IMPLEN N50-Go Nano Photo Meter User Manual
- June 10, 2024
- IMPLEN
Table of Contents
IMPLEN N50-Go Nano Photo Meter
Product Information
The product is a spectrophotometer with a software version of 4.6.16350 and a
user manual version of 4.6.2. It is manufactured by Implen, Inc. and comes
with telephone support available for customers in North and South America. The
product meets various safety and environmental standards, including the Low
Voltage
Equipment Safety Directive, Electromagnetic compatibility (EMC) directive,
Restrictions on the use of certain Hazardous Substances in Electrical and
Electronic Equipment (ROHS), and EC Directive on Waste Electrical and
Electronic Equipment (WEEE).
Product Usage Instructions
- Accessories: The product comes with standard and optional accessories. Refer to the user manual for a detailed list of accessories.
- Getting Started:
- Spectrophotometer Installation: Follow the instructions in the user manual for proper installation of the spectrophotometer.
- Safety Information: Read the safety information section in the user manual before using the product.
- Unpacking and Positioning: Follow the instructions in the user manual for proper unpacking and positioning of the product.
- Icons: Refer to the icons section in the user manual to understand the meaning of different icons displayed on the product screen.
- Buttons: Refer to the buttons section in the user manual to understand the function of different buttons on the product.
- Measurement Screens: Refer to the measurement screens section in the user manual to understand how to read and interpret different measurement screens displayed on the product.
- Methods: Refer to the methods section in the user manual to understand how to use different methods for measuring samples using the product.
NanoPhotometer® N50-Go/C40-Go
User Manual
Version 4.6.2 Software Version 4.6.16350
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 The end user of the
NanoPhotometer® Go product (“End User”) hereby takes full responsibility for
safe storage and backup of all files and/or data that may be created, saved on
or transferred from the device. End User acknowledges that it is possible that
data and/or files may be lost or damaged, and further acknowledges and agrees
that it has sole responsibility to maintain all appropriate backup of files
and data. By using the NanoPhotometer® Go device, End User hereby agrees to
these terms, and agrees that Implen shall not be held liable for any loss,
deletion or damage of any data or files for any reason, including any damages
attributable thereto.
Telephone support is available using one of the following phone numbers from
your geographic region: North and South America
Phone: +1 818 748 6400 Fax: +1 818 449-0416 Email: info@implen-go.com Website:
www.implen-go.com Implen, Inc. Unit 104 31194 La Baya Drive Westlake Village,
CA 91362 USA
Windows and Excel are trademarks of Microsoft Corporation Redmond, WA macOS is
a trademark of Apple Inc. Cupertino, CA
2
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
Declaration of conformity for the NanoPhotometer® (C40-Go/N50Go)
This is to certify that the Implen NanoPhotometer® Go conforms to the requirements of the following directives:
2014/35/EU
Low Voltage Equipment Safety Directive
2014/30/EU
Electromagnetic compatibility (EMC) directive
IEC 60529
Protection class IP20
2011/65/EU
Restrictions on the use of certain Hazardous Substances in Electrical and Electronic Equipment (ROHS)
2012/19/EU
EC Directive on Waste Electrical and Electronic Equipment (WEEE) 2003/108/EC & 2008/34/EC. By ensuring this product is disposed of correctly, you will help prevent potential negative consequences for the environment and human health, which could otherwise be caused by inappropriate waste handling of this product.
FCC 47 CFR Part15 §15.107 and §15.109
EN 301 489-1 V1.9.2
Radio and ancillary equipment for portable use (portable equipment); EUT Operating frequency range: 2.4 2.4835 GHz
EN 300 328 V1.8.1
Electromagnetic compatibility and Radio spectrum Matters (ERM); Wideband transmission systems; Data transmission equipment operating in the 2,4 GHz ISM band and using wide band modulation techniques; Harmonized EN covering the essential requirements of article 3.2 of the R&TTE Directive
EN 301 489-17 V2.2.1
Electromagnetic compatibility and Radio Spectrum Matters (ERM)
Standards to which conformity is declared, where relevant, are as follows:
IEC/EN 61010-1:2012 EN61326-1:2013
Safety requirements for electrical equipment for measurement, control, and
laboratory use. General requirements.
Electromagnetic compatibility- generic emission standard electrical equipment
for measurement, control, and laboratory use.
Signed:
Dr. Thomas Sahiri Managing Director Implen GmbH
3
N
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
NANOPHOTOMETER® N50-GO
9
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
NANOPHOTOMETER® C40-GO
10
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
INSTRUMENT REAR PANEL
To boot/shut down the NanoPhotometer® Go push shortly (< 1 second) on the
power on/off button at the back of the NanoPhotometer® Go. Note: A long push
(> 3 seconds) initiate a hard reset. Only activate a hard reset of the
NanoPhotometer® Go when necessary. To avoid unnecessary hard reboots, it is
recommended to power down the unit from the onboard touch screen by pushing on
the power button ( ) in the bottom left corner of the home screen.
INSTRUMENT BOTTOM VIEW
Model name, device serial number and FCC ID are located on the identification
plate on the bottom of the instrument.
11
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
ACCESSORIES
STANDARD ACCESSORIES
Connecting Cable
USB connecting cable to connect the NanoPhotometer® Go to a computer to
control the NanoPhotometer® Go via computer (please see page 21 Software
Installation).
NanoPhotometer® Go Power Adapter
Power adapter for the NanoPhotometer® Go. Note: Use only the power adapter
supplied with your instrument or a replacement part from the manufacturer or
your supplier.
Dust Cover
OPTIONAL ACCESSORIES
Didymium Glass Filter (C40-Go) The certified didymium glass filter can be used
to verify the wavelength and photometric accuracy of the NanoPhotometer®
C40-Go cuvette applications.
12
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
Standard Solution (N50-Go)
The NanoPhotometer® Go is recalibration free therefore it is not necessary to
review the photometric accuracy on a regular basis. If the SOPs of a
laboratory require a routine control of the photometric accuracy standard
solutions can be used.
Note: Specifications of the standard solution are guaranteed for at least one year. Please see expiration date. Once a vail is opened it can be used for 30 minutes. Note: Please read the Material Safety Data Sheet carefully prior to using this product.
DiluCellTM (C40-Go)
The DiluCellTM is a disposable cuvette with shortened path length for virtual
dilution of cuvette-based samples. Due to the reduced path length the
DiluCellTM provides an automatic dilution without the need of a physical
dilution of higher concentration samples. There are two different sizes of
DiluCells available DC 10 and DC 20 which allow an automatic 1/10 and 1/20
dilution of the sample respectively. Bypassing manual sample dilutions reduces
dilution errors and cross-contamination making DiluCellTM ideal for GLP.
Combined with small sample volume requirements and bubble free filling, the
DiluCellTM allows for convenient spectrophotometric analysis from 340 – 950
nm.
Barcode Reader
It is possible to import sample names from 1D and 2D barcodes. Connect a
compatible barcode reader to the USB port of the NanoPhotometer® Go and push
on the sample name input window. After scanning a barcode the sample name will
be shown in the input window. The imported name can then be edited or replaced
completely.
Barcode readers that have been tested and verified compatible:
1D:
Honeywell Voyager 95X0 Single-Line Laser Scanner
Datalogic Touch65
1D & 2D: AGPtEK SC36
Honeywell Xenon 1900g
DYMO Label Printer It is possible to connect a DYMO Label printer to the
NanoPhotometer® Go for direct printing on standard or cryo labels. Recommended
and tested printers are the DYMO LabelWriter 4XL/5XL (label size 10.3 x 15.8
cm) and the DYMO LabelWriter 450/550 (5.4 x 10 cm).
Cryo labels can be printed with DYMO Label printer 4XL and 450 using the
following label format: 26 x 12.7 mm and 9.5 mm circle (landscape).
13
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 Note: Cryo label paper
is not available/compatible for the DYMO Label printer 5XL and 550. Note:
After starting the NanoPhotometer® Go, plug in the DYMO printer. The home
screen will be shown. Wait at least for 30 seconds for driver installation.
Cryo labels (26 x 12.7 mm and 9.5 mm circle):
Note: Orientation of cryo labels on carrier foil needs to be landscape.
Printout for DYMO LabelWriter 4XL/5XL
Printout for DYMO LabelWriter 450/550
Note: The printouts are optimized for the label size of the DYMO LabelWriter 4XL. All other DYMO printers can be used. However, the font size will be zoomed to utilized paper size.
14
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
HP Printer
Printing from the NanoPhotometer® Go is possible via USB (HP printers) and
through network connection. Network printing should be possible with AirPrint
/ IPP compatible printers supporting PDF format. For network printer settings
see page 96 Network Printer.
Note: IPP version 2.2 is required and some printer configuration settings might need to be changed in order to allow communication with the NanoPhotometer® Go.
The following HP printers have been tested and deemed compatible to print via USB
connection:
HP LaserJet 3030
HP DeskJet 2543
HP LaserJet m1522nf MFP
HP DeskJet 1110
HP LaserJet 400 color M451nw
Further HP printers are available upon request.
Note: After starting the NanoPhotometer® Go, plug in the HP printer. The home screen will be shown. Wait at least for 30 seconds for driver installation.
CONNECTIVITY
USB A
There is a USB A port on the front as well as the rear panel of the
NanoPhotometer® Go which is compatible with standard portable USB 2.0 storage
devices (back) and USB 3.0 (front) for direct data transfer in a variety of
formats including Excel. It is also possible to connect a mouse, keyboard,
barcode reader, DYMO printer or HP printer directly to the NanoPhotometer® Go.
Note: We recommend using FAT/FAT32 formatted 2.0 USB flash drives. The USB
flash drive size is due to the standard formatting currently limited to 32 GB.
Encrypted USB flash drives are not compatible with the NanoPhotometer® Go.
Note: Cordless Bluetooth mice are not supported. Use wired mice only. Note:
Connect mouse and keyboard before starting the NanoPhotometer® Go.
USB B
There is a USB B port located on the rear panel of the instrument which is
compatible with the USB cable provided to connect the NanoPhotometer® Go to a
computer. This USB connection can be used to control the NanoPhotometer® Go
via computer.
LAN
There is an Ethernet (LAN) connection port on the rear panel of the instrument
which enables the NanoPhotometer® Go to connect with the local network. This
Ethernet connection can be used for data transfer from the NanoPhotometer® Go
to a local network, to control the NanoPhotometer® Go via a control device and
network printing. Data transfer is possible by saving on a defined network
folder (see page 95 Network Folder) or on the NanoPhotometer® Go file server
(see page 38 Data Transfer via File Server).
15
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 Note: Plug in the LAN
cable before starting the NanoPhotometer® Go. Note: The maximum LAN cable
length is 10 meters. Bit rate is 1 Gbit/s
WiFi The NanoPhotometer® Go is equipped with WiFi, which can be used as a WiFi
network or as a WiFi Hotspot. The WiFi network allows same functionality as
the Ethernet connection including direct printing via AirPrint / IPP
compatible printers supporting PDF format. Note: IPP version 2.2 is required
and some printer configuration settings might need to be changed in order to
allow communication with the NanoPhotometer® Go. The WiFi Hotspot provides the
option to control the NanoPhotometer® Go by other WiFi devices like computer,
tablets or smartphones. WiFi Hotspot connection details: SSID: NanoPhotometer®
Go serial number Password: Implenuser Note: Due to the limitations of some
handheld devices saving to a wireless device is limited to 40 measurements per
dataset. Larger datasets can be saved on the NanoPhotometer® Go itself.
HDMI There is an HDMI port located on the rear panel of the NanoPhotometer® Go
which is compatible with HDMI 1.4 cables (or better) to connect the
NanoPhotometer® Go to HDMI compatible monitors. Note: The maximum HDMI cable
length is 5 meters.
16
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
NANOPHOTOMETER® GO SPECIFICATIONS
NanoVolume Performance N50-Go
Detection Range dsDNA Detection Range BSA Sample Volume Photometric Range (10 mm equivalent) Path length Dilution Factor
5 7,500 ng/µl 0.15 217 mg/ml 0.3 2 µl 0.1 150 A
0.67 and 0.07 mm 15 and 140
Cuvette Performance C40-Go
Detection Range dsDNA
0.1 – 130 ng/µl
Detection Range BSA
0.003 – 3.7 mg/ml
Photometric Range
0 – 2.6 A
Center Height (Z-Height)
8.5 mm
Cell Types
outside dimension 12.5 x 12.5 mm
Heating
37°C ± 0.5°C
Optical Specifications Wavelength Scan Range Measure Time For Full Scan Range
Wavelength Reproducibility Wavelength Accuracy Bandwidth Absorbance
Reproducibility
Absorbance Accuracy Stray Light Optical Arrangement
C40-Go: 200 900 nm N50-Go: 200 650 nm 2.5 4.0 seconds
C40-Go: ± 0.2 nm N50-Go: ± 1nm C40-Go: ± 0.75 nm N50-Go: ± 1.5 nm C40-Go: <
1.5 nm N50-Go: < 3 nm C40-Go (cuvette): < 0.002 A @ 0 – 0.3 A @ 280 nm
CV < 1% @ 0.3 2.0 A @ 280 nm N50-Go (Lid 15): < 0.004 A (0.67 mm path) @ 280
nm
CV < 1% @ 0,3 – 1,5 A @ 280 nm < 1.75 % @ 0.7 A @ 280 nm of the reading
C40-Go: < 0.5% @ 240 nm using NaI N50-Go: < 2% @ 240 nm using NaI 1 x 4096
CMOS Array
Lamp Lifetime
Xenon flash lamp 109 flashes, up to 10 years
17
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
Processing Power and Compatibility
Operating System
Linux based OS
Onboard Processor
Intel Celeron dual core 2.4 GHz
Internal Data Storage
C40-Go, N50-Go: 32 GB
Software Compatibility
Windows 8, 10 (32 & 64 bit), OS X (Intel x86 und Apple M1), iOS and Android OS
General Specifications Main Body Size Weight Operating Voltage Display Certifications In- and Output Ports Security
200 mm x 200 mm x 120 mm 3.8 5.2 kg depending on configuration 90 250 V ± 10%, 50/60 Hz, 90 W, 18/19 VDC 1024 x 600 pixels; touchscreen glove compatible CE, IEC 61010-1:2012 and EN 61326-1:2013 2x USB A, USB B, HDMI, Ethernet, WiFi Slot for Kensington lock
Features and specifications are subject to change without notice. US Patents 20080204755 and 20080106742 Windows is a trademark of Microsoft. Mac OS & iOS are trademarks of Apple, Inc. Android OS is a trademark of Google. Linux is a trademark of Linus Torvalds.
18
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
2. GETTING STARTED
SPECTROPHOTOMETER INSTALLATION
SAFETY INFORMATION
Before commencing installation, please take time to familiarize yourself with
warning labels and symbols on your instrument and their meaning. These are to
inform you where potential danger exists or particular caution is required.
Improper use may cause personal injuries or damage to the instrument. The
instrument must only be operated by appropriately trained and experienced
personnel. Please read the complete user manual prior to use.
direct current Overvoltage catetory: Class II Maximum operating altitude: <
2000 m Polution degree: 2
Do not open the instrument as this can expose the operator to electrical
power, UV light, and delicate fiber optics or damage the instrument.
Do not use damaged power cords, accessories, and other peripherals with your
NanoPhotometer® Go. Use only the delivered and specified power supply.
Do not expose the NanoPhotometer® Go to strong magnetic, electrical fields,
water, chemicals or any type of liquid as heavy rain or moisture.
Do not put the instrument into fire, as it may swell or explode. Do not store
at or use near any type of heat source, especially temperatures above 60°C or
in an explosive atmosphere.
Do not leave your NanoPhotometer® Go on your lap or near any part of your body
to prevent discomfort or injury from heat exposure.
Do not place objects on top of the NanoPhotometer® Go.
Biological samples may contain or have the potential to transmit infectious
diseases. Be aware of the health hazard presented by such samples and wear
appropriate protective equipment. Handle such samples with the greatest of
care and according to applicable regulatory and organization requirements
before working with such potential infectious materials. Note: Do not spill
any biological samples on instrument components. If spill occurs, disinfect
the instrument immediately following your laboratory protocols and the
cleaning instruction of the instrument (see page 107 Maintenance).
The symbol on the product, or on the documents accompanying the product,
indicates that this appliance may not be treated as household waste. Instead
it shall be handed over to the applicable collection point for the recycling
of electrical and electronic equipment. Disposal must be carried out in
accordance with local environmental regulations for waste disposal.
19
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
UNPACKING AND POSITIONING
Check the contents of the package against the delivery note. If any shortages
are discovered, inform your supplier immediately. Inspect the instrument for
any signs of damage caused in transit. If any damage is discovered, inform
your supplier immediately. Ensure your proposed installation site conforms to
the environmental conditions for safe operation: indoor use or dry
environment. Note: Do not expose your NanoPhotometer® Go near liquids,
chemicals, rain, moisture or dusty environments. Working temperature range 10
– 40°C; If the cuvette heating is used the range is 10 – 27°C. Storage
temperature range is 0 – 40°C. Do not store the instrument below or above this
temperature. If the instrument is subjected to extreme temperature changes, it
may be necessary to allow the instrument to equilibrate. Turn the instrument
off and then on again once thermal equilibrium has been established (~2 – 3
hours). Maximum relative humidity (non-condensing) of 80% up to 31°C
decreasing linearly to 50% at 40°C. The instrument must be placed on a stable,
level surface that can support 4-5 kg. Ensure that air can circulate freely
around the instrument. Confirm while powered on that no materials reduce air
circulation. Avoid direct sunlight as it may bleach parts of the instrument
and can cause damage to plastic parts. The equipment should be positioned such
that in the event of an emergency the main plug can be easily located and
removed. Always carry the instrument by holding the main corpus of the
instrument and not e.g. on the optional attached display or NanoVolume
pedestal. The equipment must be connected to power with the 90W power
supply/cord supplied by Implen. The power outlet must have a protective
conductor (earth/ground). It can be used on 90-250 V ± 10%, 50-60 Hz power
supply system. Please read the complete user manual before first use. Turn the
instrument on using the power button on the rear panel after it has been
plugged in. The instrument will perform a series of self-diagnostic checks.
Please contact original supplier immediately if technical or sample handling
difficulties are experienced. Note: If this equipment is used in a manner not
specified or in environmental condition not suitable for safe operation, the
protection provided by the equipment may be impaired and the instrument
warranty voided.
20
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
SOFTWARE INSTALLATION
NPOS OVERVIEW
NPOS is a Linux-based operating system designed for the NanoPhotometer® Go.
NPOS can store data either to a common directory or be configured to save to
independent directories according to file format and/or instrument.
NPOS can save data in an Implen IDS format, PDF or as an Excel format file.
Note: PDF and Excel files cannot be opened on the NanoPhotometer® Go. Files
need to be transferred to a computer or device where Excel or a PDF reader is
available. Note: Please do not connect the instrument to a computer until the
NanoPhotometer® Go NPOS software is installed on the computer.
REQUIREMENTS AND COMPATIBILITY
The NPOS user interface is designed that all features can be operated by using
a touchscreen. If the software is installed on a computer without touchscreen,
the user interface can be operated by using keyboard and mouse. Before
starting the installation process, ensure that the software of the control
device is compatible.
Compatible Control Devices
Computer: PC: Windows 8 / Windows 10 (32 & 64 bit) Mac: macOS Catalina / Big
Sur (Intel x86 und Apple M1) Tablets (minimum requirements): iPad: iOS 13
Android (quad core 1.2 GHz with 1 GB RAM): Android version 10 Smartphones
(minimum requirements): iPhone: iOS 13 Android (quad core 1.2 GHz with 1 GB
RAM): Android version 10
Windows is a trademark of Microsoft. Mac OS & iOS are trademarks of Apple.
Android OS is a trademark of Google. Linux is a trademark of Linus Torvalds.
Note: There are two user interfaces of the software available; one for built-
in touchscreen, computer and tablets and one for smartphones.
INSTALLING SOFTWARE ON COMPUTER
The NanoPhotometer® Go software can be installed on compatible Windows and Mac
computer systems. Various operating systems and computer hardware may cause
the setup procedure to differ from that described here. This process is given
as guidance only; it may need adaptation for other systems.
Note: Do not connect the NanoPhotometer® Go to the PC/Mac before NPOS
installation. Note: If a previous version of the NPOS software is already
installed on the computer, remove the USB cable and uninstall the NPOS
software before installing the new software
21
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 version. Note: The
Windows and Mac installation files are located on the Implen USB flash drive
which is included with the NanoPhotometer® Go at time of delivery. The files
are available for free download at any time in the download area of the Implen
website (www.implen.de/downloads).
NPOS installation for single user on Windows computer 1. Check the installed
version of the NanoPhotometer® Go firmware
(Preferences/About) and update it to the latest version if necessary, before
starting the installation/update of the NPOS software on your computer. 2.
Start the NPOS installation file and follow the installation routine for
single user installation. Installation file can be found on the Implen USB
flash drive which is included with the NanoPhotometer® Go at time of delivery
or can be downloaded from the Implen webpage: www.implen.de/downloads. Full
administration rights are required for the installation. If you have
insufficient privileges, installation may fail. If in doubt consult your
computer administrator. 3. For multi user installation (only necessary for
Windows installation) choose on the License Agreement dialog the option
“Advanced” and on the following dialog “Install for all users of this machine”
4. Start the NPOS software and select the desired connection. For a
connection via USB cable, connect the NanoPhotometer® Go to the PC using the
USB cable supplied. For a connection via WiFi hotspot, ensure a stable WiFi
connection between the PC and NanoPhotometer® Go WiFi HotSpot (SSID: serial
number, password: Implenuser). For a network connection, connect the
NanoPhotometer® Go to the local network via an Ethernet cable or WiFi network
(see page 92 Network).
22
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
Note: If there is Avira installed on your computer, it is recommended to
switch off the browser safety. This may interfere with the NPOS running on
your computer.
INSTALLING NANOPHOTOMETER® GO APP ON TABLET OR SMARTPHONE
The NanoPhotometer® Go App can be installed as an application on tablets and
smartphones with compatible Android and iOS operating systems (see page 21
Requirements and Compatibility). The NanoPhotometer® Go App is available for
free download in the app store (Apple Store and Google Play Store).
1. Download and install the NanoPhotometer® Go App from the app store 2.
Connect the tablet or smartphone via WiFi HotSpot (SSID: Serial number,
password: Implenuser) or WLAN network to the NanoPhotometer® Go. 3. Open the
NanoPhotometer® Go App and choose the connection type:
4. When connected the NanoPhotometer® Go will recognize the tablet/smartphone
as a remote control device and measurements can be initiated from the tablet
or smartphone.
5. Results will be shown on the tablet or smartphone once measurements have
been taken.
Note: In order to install the NanoPhotometer® Go App on a tablet or
smartphone, the device must have an established internet connection to access
the app store for app download. Note: The version of the app and the software
of the NanoPhotometer® Go should be the same. Different versions may have not
full functionality.
FIRST STEPS AND CONFIGURATION WIZARD
When starting the Implen NPOS the first time an Implen configuration wizard is
shown. Please accept the End User License Agreement (EULA) and select the
country in which the NanoPhotometer® Go is used and confirm.
23
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
PRINTER INSTALLATION
For printers connected via USB connection: 1. Switch NanoPhotometer® Go on /
home screen 2. Connect DYMO/HP printer via USB cable 3. DYMO/HP printer is
ready to use after 30 seconds
Note: Make sure that the home screen is shown when connecting a printer. If
the USB connection between the printer and the NanoPhotometer® Go is
established while a method is open, the printer function may fail. Always
return to the home screen before connecting a printer. Note: Check printer
compatibility (page 15 HP Printer) For network printers:
1. Assure either LAN or WiFi network connection 2. Set printer IP in
preferences (page 96 Network Printer) 3. Printer is available in methods for
printing Note: Check printer compatibility (page 15 HP Printer)
24
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
3. NANOPHOTOMETER® GO BASICS
The NanoPhotometer® Go product line offers a solution for NanoVolume (N50-Go)
and standard cuvette (C40-Go) applications. NanoVolume applications start with
a minimum sample volume of 0.3 l. Standard cuvette applications can be
performed with 10 mm, 5 mm, 2 mm, 1 mm and 0.5 mm path length quartz, glass,
or plastic cuvettes with a center height of 8.5 mm.
APPLICATIONS OVERVIEW
The NanoPhotometer® N50-Go comes with two pre-programmed applications, Nucleic
Acids and Protein UV. To select a method, tap the corresponding icon and the
method opens immediately.
There is an upgrade option for the N50-Go available. The upgraded N50-Go is a
N50 Touch with access to all applications available. Homescreen C40-Go:
25
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
The following methods are available for the upgraded N50-Go and the C40-Go. *not available for N50-Go
Method Icons
Description
Nucleic Acids
Concentration, purity, and dye incorporation for DNA, RNA, Oligo, and other nucleic acids
Protein UV
Protein UV determination at 280 nm (or in a range of 200 330 nm), purity and dye incorporation
Kinetics*
Time vs. Absorbance readings
Protein Assays*
BCA (562 nm), Bradford (595 nm), Lowry (750 nm), and Biuret (546 nm) Assays
OD600*
Measures cell density at 600 nm (or in a range of 200 900 nm)
Stored Results Archive of stored results
Stored Methods Collection of stored custom methods
More Apps*
Additional applications found on a secondary method screen
Wavelength Wavescan
Define one or multiple wavelength between 200 900 nm (N50-Go: 200 650 nm)
for absorbance measurements
Define desired full scan range anywhere between 200 – 900 nm (N50-Go: 200 –
650 nm)
Concentration*
Define extinction coefficient for automatic concentration calculations
Absorbance/ Ratio*
Define two wavelengths absorbance/ratio calculation
26
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
Standard Curve* Create a standard curve at a defined wavelength
Custom Apps*
Optional custom applications for personalized methods tailored to individual spectroscopy needs
ICONS
Icon Name
Action
WiFi Network WiFi network active; status of WiFi connection
WiFi Hotspot WiFi hotspot active
Assistance
Opens the assistance page
Preferences Opens the preferences page
Home Screen Store Method Save Data
Returns to home screen with application icons for method selection.
Opens a dialog pop up with the possibility to store the actual method
parameter to a custom method
Opens a save dialog pop up
Leave Method Returns to the previous application selection
Back
Returns to the previous page (smartphone only)
Next/Confirm Confirms parameter and opens the next screen (smartphone only)
Print Data
Opens a print dialog pop up (only shown when a printer is available)
Delete Data Opens a delete dialog pop up
Parameter
Opens parameter window
Results
Opens result window
Graph
Opens graph result window
Table
Open/shows results in table format
Add Folder Manage Data Delete
Adds a new folder to the directory
Opens a dialog pop up with several action options including delete, rename or
import folders/files/data as well as copying or move folders/files/data to
defined directories
Deletes added functions in parameter; empties input windows
Full Scale Cancel
Restores graph to original size without zoom
Returns to the previous screen/closes window without implementing any changes
27
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
BUTTONS
When opening a method and starting a sample measurement, a blank measurement
is required. For the blank measurement, either water or sample buffer can be
used to give the NanoPhotometer® Go a reference of what zero should be. It is
recommended to re-apply the blank solution and measure it as a sample to
ensure the graph of the blank spectra is a flat line.
To initiate the spectral scan of the sample push the sample button. The data
will be temporarily stored until the method is exited; at this time the user
needs to define if the samples should be saved.
The auto sample button is disabled by default. When switched on sample
measurements are started automatically as soon as the lid arm is closed. Auto
sample function is only available for sample and not for blank measurements.
MEASUREMENT SCREENS
METHODS
Side Tab Bar
On the left side of the measurement screen there is a vertical tab bar that
contains four tabs including: parameters, results, graph, and table. The
different tabs allow the user to organize the measurement screen. It is
possible to show or hide the different areas on the screen. Default screen for
computer shows all areas, for the built-in screen and the tablet version the
table is hidden.
Parameter
Results
Graph
Table
Note: There is no tab bar available for smartphone versions. The parameter,
results and graph screens are shown full screen. Parameters need to be
confirmed ( ) to get to the measurement screen. It is possible to toggle
between the results and graph area by swiping left and right. There is no
table area available for smartphones.
28
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
Parameter area In the parameter area it is possible to define all necessary
parameters for a measurement as well as turn on cuvette mode and initiate
cuvette heating. The standard measurement screen shows the parameter area open
by default. The parameter area is automatically hidden when starting either a
blank or sample measurement by pushing on the Blank or Sample button. It is
also possible to hide the parameter area by tapping the parameter tab in the
vertical side tab bar.
Results area The results area shows the method specific results of the grey
highlighted measurement in the table including concentration, absorbances, and
relevant ratios. It is also possible to change the units of the calculated
concentrations in the results area with a dropdown selection menu.
Table area The table area collects the results of all samples in an active
method. The first table column shows a tick box. Selecting the samples with
the tick box the graphs are overlaid in the graph area. With the header tick
box it is possible to select/unselect all samples (maximum number of sample
selection is 30). The second column of the table indicates whether the
measurement is saved ( ) or not saved (blank field).
With the edit button the sample name of a single sample can be edited.
1. Select one sample in the table (selected sample will be highlighted in
grey)
2. Push on Edit button
3. Change sample name
4. Confirm with the “Confirm” button Note: It is not possible to edit sample
names of opened IDS files.
Graph area The graph area shows a chart with the graph of the actual
measurement or the selected line(s) in the table (tick box selection). There
is an overlay toggle switch on the left bottom of the graph area. If the
overlay option is enabled the graphs of the measurements will be automatically
overlaid. To change the overlaid graphs use the tick boxes in the table.
Note: It is only possible to overlay up to 30 graphs in a chart. If more than
30 data are selected a message will appear that says “More than 30 samples
have been selected. Only 30 will be shown in graph.” Note: The overlay button
is not available on the NanoPhotometer® Go touch screen and smartphones, only
on tablets and computer versions.
It is possible to zoom in and out any area of the diagram (x- and y-axis).
Undo the zoom by pushing the full scale icon ( ).
Note: Maximum zoom is 20 nm for the x-axis and 0.01A for the y-axis.
As legend option the sample name in the table is colored in the same color as
the graph in the chart. Pushing on the graph opens a pop up which shows the
sample name, wavelength and
29
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 absorbance of the
selected wavelength. It is possible to display the results of other graphs by
changing the sample name with the dropdown option.
DATA PROCESSING DIALOGS
PRINT
Selecting the print icon ( ) opens a full screen dialog window with various
print options. The print icon is only shown if a printer is available. The
print command is sent primarily to DYMO or HP printers if they are directly
connected to the NanoPhotometer® Go via USB cable. If no USB printer is
available, the print command is sent to the defined network printer, if
configured. Network printers can be configured in the preferences by entering
the printer IP (see page 96 Network Printer). All ticked samples are printed.
Note: If a printer is directly connected to the NanoPhotometer® via USB, this
printer will have the highest priority and will be used by default when
selecting Print on NanoPhotometer®. In order to print utilizing a printer on
the network, please disconnect the connected USB printer.
Note: The print icon is only shown, if a printer is available. Note: It is
only possible to operate one printer at a time. Do not connect more than one
printer to the NanoPhotometer® Go. Note: The print option is not available in
smartphone apps.
Auto Print If the auto print function is enabled, each measurement will be
printed directly after the measurement. Auto print function is available for
DYMO printer, HP printer connected via USB cable and network printer.
30
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 Note: Default setting
for auto print is off. If enabled in one method it is set default on for all
methods and needs to be switched off if not required. Note: The auto print
function is not available for printing via control device (local computer
printer).
Cryo Label Print To print on cryo labels connect a DYMO printer (4XL or 450)
to the NanoPhotometer® Go and insert the specified cryo label paper (26 x 12.7
mm and 9.5 mm circle / landscape mode). Note: Cryo label paper is not
available/compatible for the DYMO Label printer 5XL and 550.
SAVE
Selecting the save data icon ( ) opens a full screen overlay dialog window
with various save options.
By default all samples are ticked in the first column of the table and will be
saved. It is possible to select samples for saving by using the tick boxes.
The header tick box selects/deselects all samples. Note: In the smartphone app
there are always all measurements saved, no selection possible.
Save as Type With the Save as Type option it is possible to specify the file
type for saving. File type options include Excel, PDF and Implen Document
Source (IDS). It is possible to save different file formats simultaneously.
Note: IDS files cannot be saved on control devices. IDS files can only be
saved in the NanoPhotometer® storage, defined network folder or on a USB flash
drive. It is not possible to save IDS files from opened data. In these cases
the IDS tick box is greyed out. Note: PDF and Excel files cannot be opened on
the NanoPhotometer® Go. The files need to
31
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
be transferred to a computer or device where Excel or PDF reader is available.
Note: All data saved on the NanoPhotometer® Go are stored on an internal micro
SD card. It is recommended to back up the data regularly to a hard drive of a
computer or network. In the rare case that the micro SD card crashes data loss
cannot be ruled out.
Implen Document Source File
The Implen Document Source (IDS) file is a specific file format, which can
only be opened with the NPOS software. It is a hardcopy file which cannot be
changed. This file type contains all measurement information including raw
data, results, values, and parameters.
Note: The saved files contain only the selected samples at the time the file
is saved.
Excel File
Measurement data can be saved as Excel file. This file type contains all
measurement information including raw data, results, and parameters.
Note: The saved files contain only the selected samples at the time the file
is saved.
PDF File
Measurement data can be saved as PDF file. This file type contains all
measurement information including raw data, results, and parameters. It is
possible to configure the table columns for PDFs and printouts in preferences
(page 97 Report Configuration).
Note: The saved files contain only the selected samples at the time the file
is saved.
File Name Enter the name for the file. Allowed characters are: A…Z a…z 0…9 , .
– ( ) @ ! = _ ~ ; [ ] { } `
Note: Blank character is not allowed.
Storage Shows folder directory to select the save location. Options include:
NanoPhotometer®, USB flash drive (if connected), Network folder (if defined)
and Control Device. If Control Device is selected the data will be transferred
to the control device that is currently connected with the NanoPhotometer® Go.
Note: It is not possible to save IDS files to a control device like computer
(PC/Mac), tablets or smartphones.
Auto Save To prevent data loss, all measurements are automatically stored as
IDS file on the internal memory of the NanoPhotometer® Go. These backup copies
can be found in the Autosave folder of the NanoPhotometer® (Stored
Results/NanoPhotometer/Autosave) for up to ten days. Files contain the base
name Backup, the method name, and a time/date stamp. After ten days the
autosave files are automatically moved to an autosave archive folder. The
autosave archive folder can only be accessed via NanoPhotometer® Go file
server (see
32
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
page 38 Data Transfer via File Server). Data in the autosave archive folder
are not automatically deleted.
The content of the Autosave Archive folder can be deleted via the action button Results:
in Stored
DELETE
Selecting the delete icon ( ) opens a full screen overlay dialog window. All
data which are selected (tick) in the first table column will be deleted. The
header tick box selects/deselects all samples. Initiate the deletion with the
delete button. Confirm the deletion in the following warning message: “Do you
want to delete all/selected files?” select cancel (x) will return to the
delete menu screen or confirm with delete to delete the selected data. Note:
The delete function is not available for the software version designed for
smartphones.
33
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
STORE METHODS
With the favorite icon it is possible in each method to save the parameter
settings for easy access of custom defined methods. Select the desired
parameter settings and open the Stored Method dialog by pushing o the favorite
icon .
Enter a method name and select a save location in the folder. Options include:
NanoPhotometer®, USB flash drive (if connected) and Network folder (if
defined). Push the store button to save the method. Stored Methods can be
opened on the homescreen by opening Stored Methods (see page 87 Stored
Methods).
BASIC OPERATION
The NanoPhotometer® Go product line offers a solution for NanoVolume (N50-Go)
and standard cuvette (C40-Go) applications. NanoVolume applications start with
a minimum sample volume of 0.3 l. Standard cuvette applications can be
performed with 10 mm, 5 mm, 2 mm, 1 mm and 0.5 mm path length quartz, glass,
or plastic cuvettes with a center height of 8.5 mm.
NANOVOLUME MEASUREMENT BASICS (N50-GO)
1. Select a method depending on your sample and set the parameters for the
measurement.
2. Ensure that the sample window on pedestal and the mirror in the lid arm
are clean. 3. Raise the lid arm and pipette the appropriate amount of blank
solution onto the
illuminated sample window on pedestal. The Illumination turns automatically
off when the lid arm is lowered.
34
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
Note: Do not overfill the well. Note: The low energy red light (LED)
illumination can be switched of in Preferences 4. Lower the lid arm and
initiate a blank measurement with the blank button 5. Clean the measurement
window and mirror on the lid arm with a slightly wet lint-free tissue. Use
water, 70% ethanol or isopropanol if needed.
Note: Make sure that the metal contact face (around the measurement window and
the mirror) is clean. Note: Do not use aggressive solvents such as strong
acids or bases or organic solvents at any time (see page 38 Solvent
Compatibility (N50-Go). If unsure please contact support@implen-go.com for
detailed information about your specific reagent/solvent. 6. It is possible to
enter a sample name for each sample in the input window “enter sample name”.
Note: Allowed characters are: A…Z a…z 0…9 , . – ( ) @ ! = _ ~ ; [ ] { } `
blank character 7. Raise the lid arm and pipette the appropriate amount of
sample solution onto the illuminated sample window. Upon completion of
measurement raise lid arm, clean the surfaces and apply the next sample. Note:
Parameter setting Volume 1 – 2 µl adjusts the path length automatically. The
parameter setting Volume 0.3 µl measures only the 0.07 mm path length for
higher concentrations (dsDNA > 420 ng/µl / BSA > 12.6 mg/ml). Note: The sample
window on pedestal must be clean and residual fluff from any cleaning wipe
must be removed for optimum performance.
CUVETTE MEASUREMENT BASICS (C40-GO)
The NanoPhotometer® Go is compatible with standard cuvettes having an 8.5 mm
center height. The light path is indicated with two white arrows.
35
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
1. Select a method depending on your sample and set the parameters for the
measurement.
2. Add a blank solution to a cuvette and ensure that the filling volume is
sufficient to allow light path to pass through the solution.
3. Insert cuvette into the cell holder. 4. Initiate a blank measurement with
the blank button. Upon completion of measurement
remove the cuvette. 5. Add sample to a cuvette and ensure that the sample
volume is sufficient to allow light to
pass through the sample. 6. It is possible to enter a sample name for each
sample in the input window “enter sample
name”. Note: Allowed characters are: A…Z a…z 0…9 , . – ( ) @ ! = _ ~ ; [ ] { }
` blank character 7. Initiate a sample measurement with the sample button.
Upon completion of measurement remove the cuvette. 8. Apply further samples.
36
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
SAMPLE HANDLING TIPS
NanoVolume Methods (N50-Go)
The sample window on pedestal is illuminated (N50-Go) with a low energy red
light to assist with accurate sample application. The red light is switched
off once the lid arm is closed. It is possible to disable the illumination
feature in preferences of the NPOS software.
The minimum volume that can be used for NanoVolume samples is 0.3 µl (N50-Go
dsDNA > 420 ng/µl and BSA > 12.6 mg/ml). For automatic path length setting at
least 1µl is needed.
The maximum volume that can be used for NanoVolume samples is 2.0 µl (N50-Go).
The sample can be fully recovered after measurement with a pipette if desired.
Note: Minimal cross contamination cannot be avoided on molecular level. Proper
cleaning is important to ensure accurate measurements. In most cases a dry
lint-
free laboratory wipe is sufficient to clean the sample quartz surfaces. In the
case of highly concentrated samples or certain proteins, the recommended
procedure for cleaning is to use a slightly wet lint-free laboratory wipe
(with water or 70% EtOH depending on sample type) to thoroughly clean the
sample surface. It is mandatory that the metal contact face around the
measurement window and the mirror is clean. Cuvette Methods (C40-Go)
The cuvette holder is compatible with standard 10 mm path length quartz, glass
and plastic cuvettes with an optical height of 8.5 mm.
It is also possible to use cuvettes with 5 mm, 2 mm, 1 mm or 0.5 mm path
lengths, but there may be an adapter necessary. Please ask your cuvette
supplier for a suitable adapter.
The minimum volume for accurate measurements depends on the cuvette type used;
it is necessary that the light passes through the sample for accurate
measurements. Center height is 8.5 mm.
Note: The cuvette holder is not removable. Do not pour any cleaning solution
into the cuvette holder as larger amounts of liquids can get into the
instrument and cause damage.
37
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
SOLVENT COMPATIBILITY (N50-GO)
Most solvents typically used in life science laboratories are compatible with
the NanoPhotometer® Go NanoVolume sample surfaces. The following solvents are
compatible for use with the NanoPhotometer® models N50-Go at room temperature:
Acetone ( 5%)
Methylene chloride
Acetonitrile
MOPS
Benzene
Phenol (1%)
Butanol
N-propanol
Carbon tetrachloride
Toluene
Chloroform
Phosphate containing buffers
Ethanol
PBS (pH 4-10)
Ether
Citrate
HEPES
Borate
Hexane Isopropanol MES
Chloride salts
Acids > pH 2 Bases < pH 10
Methanol
Note: Highly concentrated acids and bases are not recommended. It is recommended to
wipe the sample surface with a lint-free laboratory wipe immediately upon completion of
each measurement. For more information about compatibility of specific solvents not listed
above, please contact the Implen support team (support@implen-go.com) to check the
compatibility.
DATA TRANSFER VIA FILE SERVER
All data saved on the NanoPhotometer® Go can easily be accessed from and
transferred to a computer via the NanoPhotometer® Go file server. Connection
options are LAN/WLAN, USB cable or WiFi Hotspot. It is possible to create user
accounts for password secured file server access. User accounts for file
server access can be activated in Preferences see page 94 File Server Access.
File Server Access via LAN/WLAN
For the file server access via LAN/WLAN it is necessary that both the computer
and the NanoPhotometer® Go are connected to the same LAN/WLAN network. For
connection of the NanoPhotometer® Go to LAN/WLAN see page 92 Network.
For Windows computer open the Windows explorer and enter the serial number or
the NanoPhotometer® Go IP in the address bar of the Windows Explorer (e.g.
\M80798 or
\Assigned IP Address).
38
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 Note: Serial number
and IP address of the NanoPhotometer® Go can be found in the NanoPhotometer®
Go software under Preferences/General/About. For a MAC computer open the
“Connect to Server” dialog in the “Go” menu of the Mac OS X Finder and enter
the NanoPhotometer® Go serial number or the active NanoPhotometer® Go IP
address in the server address field to connect.
Note: Serial number and IP address of the NanoPhotometer® Go can be found in
the NanoPhotometer® Go software under Preferences/General/About.
File Server Access via USB cable For file server access via USB cable
connection, connect the NanoPhotometer® Go with a USB A/B cable to the
computer and open the Windows Explorer or Connect to Server option for Mac
(see file server access via LAN/WLAN) and enter \192.168.7.1 for connection.
File Server Access via WiFi Hotspot For file server access via WiFi Hotpot the
WiFi Hotspot needs to be active on the NanoPhotometer® Go. For activation see
page 93
WLAN Settings. The computer needs to be connected to the NanoPhotometer® Go
WiFi Hotspot (SSID: NanoPhotometer® Go serial number; password: Implenuser).
Open the Windows Explorer or Connect to Server option for Mac (see file server
access via LAN/WLAN) and enter \192.168.8.1 for connection.
39
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
4. NANOPHOTOMETER® GO APPLICATIONS
The NanoPhotometer® Go comes with pre-programmed applications. For the N50-Go
are only the Nucleic Acid und Protein UV method available. There is an N50
upgrade option available for full applicational access. Any application method
can be selected by tapping the icon once or clicking on the icon (computer
based software).
NUCLEIC ACIDS
METHOD OVERVIEW
Nucleic acids in solution absorb light with a peak in the ultraviolet region
of 260 nm. For determination of nucleic acid concentration in solution the
absorbance at wavelength 260 nm is used along with the Beer-Lambert law. In
addition to calculating concentrations of nucleic acids, absorbance
measurements are also useful for estimating purity of nucleic acids by
calculating the 260/280 nm and 260/230 nm ratios. Further, it is possible to
determine the degree of labeling of nucleic acids with probes including
fluorescent dyes. Sample ControlTM gives useful information about sample
conditions. It recognizes air bubbles, sample impurities, turbidity, lint
residues and potential contaminations. If Sample ControlTM detects any
inconstancy an alert icon is shown in the result/table area. A push on the
alert icon shows additional information about the inconstancy.
MEASUREMENT PROTOCOL
1. Select the Nucleic Acids icon on the home screen.
2. To change the nucleic acid type push on dsDNA and a list with available
options opens on the right side. Options are: dsDNA, ssDNA, RNA miRNA, miRNA
Sequence, Oligo, Oligo Sequence and Custom (see Table 1 on page 43). It is
possible to enter a name for the custom nucleic acid factor for documentation.
40
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 3. For NanoVolume
application (N50-Go):
Select the volume of sample to be applied.
Note: 1 – 2 µl (default): automatic path length setting; 0.3 µl measures only
the 0.07 mm path length (for samples with concentrations > 420 ng/µl dsDNA)
For cuvette application (C40-Go): Select the path length depending on the used
cuvette. Options are: 0.5 mm, 1 mm, 2 mm, 5 mm and 10 mm
If it is desired to heat the sample to 37°C use the toggle switch to turn on
cell holder heater. When the cuvette holder has reached 37°C the toggle color
changes to green. Note: Only available for cuvette applications (C40-Go). 4.
Select the Units in which the concentration should be calculated. Options are
ng/µl (default), µg/µl and µg/ml. pmol/µl if a nucleic acid sequence is
entered for nucleic acid factor calculation.
5. The background correction is enabled at 320 nm by default. Selection
options are 320 nm, 340 nm or any wavelength in the range of 220 350 nm. The
background correction can be disabled with the toggle switch.
6. Air bubble recognition is disabled by default. Setting to on detects air
bubbles, lint residues and bad sample conditions of the sample. Note: Lint
residues and bad sample conditions are detected even if the air bubble
recognition is set to off.
41
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 7. For dye labelled
samples tick the dye labels in the list, which should be used for result
calculation. Note: If the used dye is not available in the list, please go to
preferences and add a custom dye to the dye list. There is an option for dye
correction which can be enabled/disabled with a toggle switch. Note: Dye
correction is only available for single dye selection.
8. Option to set/calculate a dilution factor for manual diluted samples.
9. Apply the blank ddH20 or buffer to the illuminated sample window on the
pedestal and close the lid arm for the reference measurement and select blank
to initiate the reading. Note: The illumination of the sample window can be
switched off in the preferences.
42
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
10. Use a lint-free laboratory wipe to clean both the sample window on
pedestal and mirror in lid arm prior to applying the next sample.
Note: It could be helpful to apply the blank a second time and read it as a
sample to ensure a proper blank.
11. Apply sample to the sample window on pedestal and press the sample button to initiate the measurement.
CALCULATIONS
Nucleic Acid Concentration
To determine the concentration of nucleic acids in solution, the absorbance is
measured at a wavelength of 260 nm. The function describing the relationship
between concentration and absorbance is a modification of the Beer-Lambert law
equation. The concentration of the nucleic acid samples can be calculated with
or without background correction depending on enabled/disabled background
correction parameter.
Without background correction:
C = A260 Factornuc With background correction: C = (A260 – ABKG) Factornuc
C
Concentration in ng/µl
A260 ABKG
Absorbance at 260 nm (10 mm path) Absorbance at selected background wavelength (10 mm path) Manual dilution factor
Factornuc Nucleic acid factor in ng*cm/µl
Table 1. Nucleic acids extinction coefficients (nuc)
Type dsDNA ssDNA RNA miRNA Oligo miRNA Seq. Oligo Seq. Custom
Factornuc 50 ngcm/µl 37 ngcm/µl 40 ngcm/µl 33 ngcm/µl 33 ngcm/µl calculated via extinction coefficient of constituent nucleotides entered calculated via extinction coefficient of constituent nucleotides entered Option to enter any factor between 15 and 150 ngcm/µl
43
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
Dye-labeled Nucleic Acid Concentration For dye-labeled nucleic acids, the
concentration of the nucleic acid is calculated using a modified form of the
Beer-Lambert equation. For these calculations, the instrument considers the
absorption maximum of the dye and a certain dye-specific correction factor at
260 nm (see Table 2 on page 46). The concentration of a dye-labeled nucleic
acid is calculated with or without background/dye correction as follows:
With background and with dye correction:
C = [(A260 – ABKG) – (cfdye (Amax, dye – ABKG))] Factornuc
With background and without dye correction: C = (A260 – ABKG) Factornuc
Without background and with dye correction: C = [A260 – (cfdye Amax, dye)]
Factornuc Without background and without dye correction:
C = A260 Factornuc
C
Concentration in ng/µl
A260
Absorbance at 260 nm (10 mm path)
ABKG Absorbance at selected background wavelength (10 mm path)
Amax, dye Absorbance value at the absorbance maximum of the dye (10 mm path)
Factornuc Nucleic acid factor in ng*cm/µl
cfdye
Dye-dependent correction factor at 260 nm
Manual dilution factor
Note: Dye correction is only available for single dye selection.
Dye Concentration
For dye-labeled nucleic acids, the concentration of the dye is calculated
using a modified form of the Beer-Lambert equation. For these calculations,
the instrument considers the absorption maximum of the dye, the dye-specific
extinction coefficient (see Table 2 on page 46). The dye concentration is
calculated with or without background correction as follows:
With background correction:
C
=
(Amax, dye – dye*
ABKG) 10-6
Ð
Without background correction:
C
=
Amax, dye mdye 10-6
44
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
C Amax, dye ABKG mdye
Concentration in ng/µl Absorbance value at the absorbance maximum of the dye
(10 mm path)
Absorbance at 320 nm (10 mm path) Molar extinction coefficient of dye in
M-1*cm-1
Manual dilution factor
Frequency of Incorporation (FOI)
FOI is the degree of labeling based on dye incorporation in a labeled nucleic
acid sample. It is generally expressed as the number of dye molecules
incorporated per 1000 nucleotides. FOI can be calculated with or without
background/dye correction as follows:
With background correction with dye correction:
FOI =
324.5 * (Amax, dye – ABKG)
dye 10-6 (A260 – ABKG – cfdye (Amax, dye – ABKG)) Factornuc
With background correction without dye correction:
FOI
=
324.5 mdye 10-6
- (Amax, dye – ABKG) (A260 – ABKG) * Factornuc
Without background correction and with dye correction:
FOI
=
mdye *
324.5 Amax, dye 10-6 (A260 – cfdye * Amax, dye)
- Factornuc
Without background correction and without dye correction:
FOI
=
324.5 Amax, dye mdye 10-6 A260 Factornuc
FOI
A260 ABKG Amax, dye mdye
Factornuc cfdye
Frequency of Incorporation (dye per 1,000 bases) Absorbance at 260 nm (10 mm
path) Absorbance at selected background wavelength (10 mm path) Absorbance
value at the absorbance maximum of the dye (10 mm path) Extinction coefficient
of dye in M-1cm-1
Nucleic acid factor in ngcm/µl Dye-dependent correction factor at 260 nm
45
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
Table 2. Dye Types, Absorbance Max, Extinction coefficient, and dye-dependent correction factors
NanoPhotometer® Go Model
N50-Go/C40-Go N50-Go/C40-Go N50-Go/C40-Go N50-Go/C40-Go N50-Go/C40-Go
N50-Go/C40-Go N50-Go/C40-Go C40-Go C40-Go C40-Go N50-Go/C40-Go N50-Go/C40-Go
C40-Go C40-Go N50-Go/C40-Go N50-Go/C40-Go N50-Go/C40-Go C40-Go C40-Go C40-Go
N50-Go/C40-Go
Dye Type
Absorbance maximum of Dye (nm)
Molar ext. coeff.
of dye mdye in M-1*cm-1
Alexa Fluor 350
346
Alexa Fluor 488
495
Alexa Fluor 532
532
Alexa Fluor 546
554
Alexa Fluor 555
555
Alexa Fluor 568
578
Alexa Fluor 594
590
Alexa Fluor 647
650
Alexa Fluor 660
663
Alexa Fluor 680
679
Cy3
550
Cy3.5
581
Cy5
649
Cy5.5
675
Oyster-500
503
Oyster-550
553
Oyster-556
560
Oyster-645
649
Oyster-650
653
Oyster-656
660
Texas Red
603
19,000 71,000 81,000 112,000 150,000 91,300 90,000 239,000 132,000 184,000 150,000 150,000 250,000 250,000 78,000 150,000 155,000 220,000 200,000 200,000 112,000
Dye-dependent correction factor at 260 nm cfdye
0.25 0.30 0.24 0.21 0.08 0.45 0.43 0.00 0.00 0.00 0.08 0.08 0.05 0.05 0.29
0.05 0.03 0.05 0.04 0.04 0.23
Ratios
Reactions utilizing nucleic acids often require minimum purity standards.
Common contaminants of nucleic acid samples include: proteins, organic
compounds, and other. Based on the common contaminants of nucleic acid
samples, the 260/280 and 260/230 ratios are calculated for nucleic acids to
give an indication of the purity of the samples. Pure DNA and RNA preparations
have expected 260/280 ratios of 1.8 and 2.0 respectively. An elevated
absorbance at 230 nm can indicate the presence of impurities as well; 230 nm
is near the absorbance maximum of peptide bonds and also indicates buffer
contamination since TRIS, EDTA and other buffer salts absorb at 230nm. When
measuring RNA samples, the 260/230 ratio should be > 2.0; a ratio lower than
this is generally indicative of contamination with guanidinium thiocyanate, a
reagent commonly used in RNA purification and which absorbs over the 230-260
nm range. If a ratio is detected out of the acceptable
46
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 range an alert icon is
shown in the results/table area. A push on the alert icon shows additional
information. The ranges for acceptable ratio values can be defined in
preferences. The ratios are calculated with or without background correction
according to if the background correction is activated during the measurements
or not as follows: Without background correction: 260/280 ratio = A260
A280
260/230 ratio = A260
A230
With background correction: 260/280 ratio = A260 – ABKG
A280 – ABKG
260/230 ratio = A260 – ABKG
A230 – ABKG
47
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
PROTEIN UV
METHOD OVERVIEW
The Protein UV method exploits the inherent absorbance of proteins at 280 nm
in combination with the Beer-Lambert Law, where each protein is characterized
by a protein specific extinction coefficient () which can be used to determine
total protein concentration of a solution. The intrinsic absorbance of
proteins is due to the presence of aromatic amino acids in their structure,
primarily tryptophan and tyrosine, as well as cysteine (oxidized cysteine
residues in a disulphide bond). The aromatic amino acid residues in a protein
containing tryptophan and tyrosine exhibit strong intrinsic absorbance at 280
nm, with a lesser contribution by phenylalanine. Therefore, it is the aromatic
amino acid residues which dictate the extinction coefficient at 280 nm for a
protein.
The most straightforward method to determine concentration of a purified,
homogenous protein with a known extinction coefficient () is by direct
measurement of UV280 provided as long as the protein contains no prosthetic
groups with strong absorption in the same region. However, for unknown
proteins including homogenous protein mixtures, it is possible to make direct
A280 measurements using a composite value derived from comparison of many
proteins, although this will only provide an approximate but close estimate of
the true protein concentration.
The NanoPhotometer® Go determines protein concentration by performing
calculations based on specific values, either pre-programmed in the instrument
or entered manually by the user. Extinction coefficient () values at 280 nm
vary greatly for different proteins due to their particular aromatic amino
acid content. Fixed values are pre-programmed in the software for certain
proteins (see Table 3 on page 52). However, if the protein of interest is not
included in the pre-programed methods it is also possible to manually enter
the specific for the protein of interest using the custom Mol. Ext.
Coefficient, custom Ext. Coefficient or custom 1/ protein factor option. For
correct calculation, it is necessary to supply either: a) the molar extinction
coefficient (M in M-1cm-1) and the molecular weight expressed in molar mass
units (g/mol); b) the mass extinction coefficient ( in l/gcm) or c) the
protein factor 1/ of the protein.
To determine the degree of dye labelling of a protein, the absorbance measured
at the wavelength corresponding to the absorbance maximum of the fluorescence
dye is used (see Table 3 on page 52). The corresponding extinction coefficient
of the dye is used along with the Beer-Lambert Law to determine the dye
concentration.
Note: It is important to ensure the extinction coefficient and units entered
are correct in order to ensure that calculations are performed properly for
accurate concentration values.
Sample ControlTM gives useful information about air bubbles, sample
impurities, turbidity, lint residues and potential contaminations. If the
Sample ControlTM detects any inconstancy an alert icon is shown in the
result/table area. A push on the alert icon shows additional information about
the inconstancy.
48
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
MEASUREMENT PROTOCOL
1. Select the Protein UV icon on the home screen.
2. To change the protein type push on BSA and a list with available options
opens on the right side. Options are: BSA, SA Mouse, SA Human, IgG Mouse, IgG
Human, IgE Human, Lysozyme, OD1, Custom (Molar Extinction Coefficient), Custom
(Extinction Coefficient) and Custom (1/). For Custom (Mol. Ext. Coefficient)
enter Molecular Weight in g/mol and the Mol. Ext. Coefficient in M-1cm-1 For
Custom (Ext. Coefficient) enter the Ext. Coefficient in l/gcm For Custom (1/)
enter the calculated protein factor 1/
3. For NanoVolume application (N50-Go):
Select the volume of sample to be applied.
Note: 1-2 µl (default): automatic path length change; 0.3 µl measures only the
0.07 mm path length (possible for samples with concentrations e.g. BSA > 12.6
mg/ml) For cuvette application (C40-Go): Select the path length depending on
the used cuvette. Options are: 0.5 mm, 1 mm, 2 mm, 5 mm and 10 mm
49
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
If it is desired to heat the sample to 37°C use the toggle switch to turn on
cell holder heater. When the cuvette holder has reached 37°C the toggle color
changes to green. Note: Only available for cuvette applications (C40-Go) 4.
The wavelength for protein measurements can be changed in the range of 200
330nm depending on the wavelength peak of the protein. Default setting is 280
nm 5. Select the Units in which the concentration should be calculated.
Options are ng/µl, µg/µl, µg/ml and mg/ml (default).
6. The background correction is enabled at 320 nm by default. Selection
options are 320 nm, 340 nm or any wavelength in the range of 220 350 nm. The
background correction can be disabled with the toggle switch.
7. Air bubble recognition is disabled by default. When enabled it detects air
bubbles, lint residues and poor conditions of the sample. Note: Lint residues
and bad sample conditions are detected even if the air bubble recognition is
set to off.
8. For dye labelled samples, tick the dye labels in the list which should be
used for result calculation. Note: If the used dye is not available in the
list, please go to preferences and add a custom dye to the dye list. 50
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 There is an option for
dye correction which can be enabled/disabled with a toggle switch.
Note: Dye correction is only available for single dye selection.
9. Option to set/calculate a dilution factor for manual diluted samples.
10. Apply the blank ddH20 or buffer to the illuminated sample window on
pedestal for the reference measurement and select blank to initiate the
reading. Note: The illumination of the sample window can be switched off in
the preferences.
11. Use a lint-free laboratory wipe to clean both the sample window on
pedestal and mirror in the lid arm prior to applying the next sample. Note: It
can be helpful to apply the blank a second time and read it as a sample to
ensure a proper blank.
51
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
12. Apply sample to the sample window on pedestal and press the sample button to initiate the measurement.
CALCULATIONS
Protein UV280 Concentration
The protein concentration in the Protein UV method is calculated with the
absorbance value of the sample at 280 nm or entered wavelength in the range of
220-350 nm along with the extinction coefficient defined by the user. The
protein concentration is calculated with or without background correction as
follows:
With background correction: C = (A280 – ABKG) Factorprot Ð
Without background correction:
C = A280 Factorprot Ð
C
Concentration (mg/ml)
A280
Absorbance at 280 nm (10 mm path) or entered wavelength
ABKG Absorbance at selected background wavelength (10 mm path)
Factorprot Protein factor in g*cm/l (1/Ext. Coeff. or MW/Mol.Ext. Coeff.)
Ð
Dilution factor
Table 3. Protein extinction coefficients (prot)
Type BSA SA Mouse SA Human IgG Mouse IgG Human IgE Human Lysozyme OD1
Factorprot [ g* cm/l] 1.499 1.493 1.718 0.714 0.735 0.654 0.379 1.000
Ext. Coeff. [l/g*cm] 0.6670 0.6700 0.5820 1.4000 1.3600 1.5300 2.6400 N/A
Mol. Ext. Coeff. [M-1*cm-1 ] 44,289 44,220 40,370 224,000 204,000 290,700 37,984 N/A
MW [g/mol] 66,400 66,000 69,365 160,000 150,000 190,000 14,388 N/A
Dye-labeled Protein UV280 Concentration
For dye-labeled proteins, the concentration of the protein is calculated using
a modified form of the Beer-Lambert equation. For these calculations, the
instrument considers the absorption maximum of the dye, and a certain dye-
specific correction factor at 280 nm (see Table 4 on page 54). The dye
concentration is calculated with or without background/dye correction for are
as follows:
With background and with dye correction:
C = [A280 – ABKG – (cfdye (Amax, dye – ABKG))] Factorprot * Ð
52
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
With background and without dye correction: C = (A280 – ABKG) Factorprot Ð Without background and with dye correction:
C = (A280 – ( cfdye Amax, dye)) Factorprot * Ð
Without background and without dye correction:
C = A280 Factorprot Ð
C
A280 ABKG Amax, dye Factorprot cfdye Ð
Concentration in mg/ml Absorbance at 280 nm (10 mm path) Absorbance at selected background wavelength (10 mm path) Absorbance value at the absorbance maximum of the dye (10 mm path) Protein factor in g*cm/l (1/Ext. Coeff. or MW/Mol.Ext. Coeff.) Dye-dependent correction factor at 280 nm Dilution factor
Dye Concentration
For dye-labeled proteins, the concentration of the dye is calculated using a modified form of the Beer-Lambert equation. For these calculations, the instrument considers the absorption maximum of the dye, and a dye-specific extinction coefficient (see Table 4 on page 54). The dye concentration is calculated with or without background correction for are as follows:
With background correction:
C
=
((Amax, dye – ABKG) mdye * 10-6
- Ð)
Without background correction:
C
=
(Amax, dye Ð) dye 10-6
C Amax, dye ABKG mdye Ð
Concentration Absorbance at the max dye absorbance value (10 mm path)
Absorbance at selected background wavelength (10 mm path) Extinction
coefficient of dye in M-1*cm-1
Dilution factor
Degree of Labeling (DOL)
DOL is the degree of labeling based on the average number of dye molecules
coupled to a protein molecule. The degree of labeling can be determined from
the absorption spectrum of the labeled antibody with or without background/dye
correction as follows:
53
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
With background and with dye correction:
DOL =
(Amax, dye – ABKG) * mprot
((A280 – ABKG) – cfdye * (Amax, dye – ABKG)) mdye
With background and w/o dye correction:
DOL =
(Amax, dye (A280 –
– ABKG mprot ABKG) mdye
W/o background and with dye correction:
DOL =
(A280
Amax, dye mprot – cfdye Amax, dye) * mdye
W/o background and w/o dye correction:
DOL =
Amax, dye mprot A280 mdye
DOL
Amax, dye
A280 ABKG mdye mprot cfdye
Degree of labeling/dye per protein ratio Absorbance value at the absorbance
maximum of the dye (10 mm path)
Absorbance at 280 nm (10 mm path) Absorbance at selected background wavelength
(10 mm path) Extinction coefficient of dye in M-1cm-1 Molar extinction
coefficient of protein (M-1 cm-1)
Dye-dependent correction factor at 280 nm
Table 4. Dye Types, Absorbance Max, Extinction coefficients, and dye-dependent correction factors
NanoPhotometer® Go Dye Type Models
N50-Go/C40-Go N50-Go/C40-Go N50-Go/C40-Go N50-Go/C40-Go N50-Go/C40-Go N50-Go/C40-Go N50-Go/C40-Go N50-Go/C40-Go C40-Go C40-Go C40-Go N50-Go/C40-Go C40-Go C40-Go
Alexa Fluor 350 Alexa Fluor 405 Alexa Fluor 488 Alexa Fluor 532 Alexa Fluor 546 Alexa Fluor 555 Alexa Fluor 568 Alexa Fluor 594 Alexa Fluor 647 Alexa Fluor 680 Alexa Fluor 790 Cy3 Cy5 DyLight 649
Absorbance maximum of Dye (nm)
346 401 495 532 554 555 578 590 650 679 785 550 649 654
Molar ext. coeff. of Dye mdye in M-1*cm-1
19,000 34,000 71,000 81,000 112,000 150,000 91,300 90,000 239,000 184,000
260,000 150,000 250,000 250,000
Dye-dependent correction factor at 280 nm cfdye
0.19 0.70 0.11 0.09 0.12 0.08 0.46 0.56 0.03 0.05 0.08 0.05 0.05 0.04
54
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
N50-Go/C40-Go
DyLight 488
493
70,000
0.15
N50-Go/C40-Go
FITC
494
70,000
0.30
N50-Go/C40-Go
Pacific Blue
409
30,000
0.20
N50-Go/C40-Go
Pacific Orange
397
24,500
0.60
N50-Go/C40-Go
pHrodo Green
505
75,000
0.20
N50-Go/C40-Go
pHrodo Red
560
65,000
0.12
N50-Go/C40-Go
r-PE
566
1,863,000
0.17
N50-Go/C40-Go
Texas Red
595
80,000
0.18
Ratios
Protein samples e.g. from whole cell lysates may contain nucleic acids. To
check the purity of the isolated protein, the 260/280 ratio is calculated to
give an indication of the nucleic acid contamination. A pure protein
preparation has an expected 260/280 ratios of 0.57. If a ratio is detected out
of the acceptable range an alert icon is shown in the results/table area. A
push on the alert icon shows additional information. The ranges for acceptable
ratio values can be defined in preferences. The ratio is calculated with or
without background correction according to if the background correction is
activated during the measurements or not as follows:
Without background correction:
260/280 ratio = A260
A280
With background correction:
260/280 ratio = A260 – ABKG
A280 – ABKG
PROTEIN ASSAYS
Available only for C40-Go, N50-Go needs upgrade for activation of Protein
Assays.
METHOD OVERVIEW
Protein concentration may be measured using colorimetric assays, in which
certain reagents are added to the protein solution to generate a colored
product; either a protein-cupric ion chelate as in the Biuret, Lowry, BCA
assays or a protein-dye complex as in the Bradford assay. In these
colorimetric assays, the absorbance is measured in the visible range at the
appropriate wavelength for each assay and compared against a standard curve
prepared by serial dilution of a protein standard of known concentration. A
linear, linear zero or 2nd order regression analysis of the calibration
standard data points is calculated by the NanoPhotometer® Go. A correlation
coefficient (R2) in the range of 0.95 to 1.00 indicates a good fit to a
straight line.
55
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
Bradford Assay Method depends on quantifying the binding of a dye, Coomassie
Brilliant Blue, to an unknown protein and comparing this binding to that of a
standard curve prepared from a set of known protein of known concentrations at
595 nm. This standard is usually BSA (bovine serum albumin).
Biuret Assay Method depends on a reaction between cupric ions and peptide
bonds in an alkali solution, resulting in the formation of a complex absorbing
at 546 nm.
BCA Assay Method depends on a reaction between cupric ions and peptide bonds
coupled with the detection of cuprous ions using bicinchoninic acid (BCA),
giving an absorbance maximum at 562 nm. The BCA process is less sensitive to
the presence of detergents used to solubilize membranes.
Lowry Assay Method is based on the Biuret reaction. Under alkaline conditions
the divalent copper ion forms a complex with peptide bonds in which it is
reduced to a monovalent ion. Monovalent copper ion and the radical groups of
tyrosine, tryptophan, and cysteine react with Folin reagent to produce an
unstable product that becomes reduced to molybdenum/tungsten blue. The bound
reagent changes color from yellow to blue. This binding is compared with that
obtained with a standard protein at 750 nm; this is usually BSA (bovine serum
albumin). Note: Detailed protocols are customarily supplied with these assay
kits, and must be closely followed to ensure that accurate results are
obtained.
MEASUREMENT PROTOCOL
1. Select protein assays icon from home screen.
2. To change the assay type click on Bradford and a list with available
options opens on the right side. Options are: BCA Assay, Biuret Assay (not for
N50 Go), Bradford Assay, Lowry Assay (not for N50-Go)
3. For Cuvette Application (C40-Go): Select the path length depending on the
used cuvette. Options are: 0.5 mm, 1 mm, 2 mm, 5 mm and 10 mm
56
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
If it is desired to heat the sample to 37°C use the toggle switch to turn on
cell holder heater. When the cuvette holder has reached 37°C the toggle color
changes to green. Note: Only available for cuvette applicatons (C40-Go). For
NanoVolume Application (only with N50-Go Upgrade): Select the dilution
depending on the sample concentration
Note: There is no automatic path length setting available in this method.
Select either a virtual dilution of 15 (path length 0.67 mm) or of 140 (path
length 0.07 mm) depending on your sample concentration. 4. Default values for
the baseline correction are depending on selected protein assay type and
instrument version: BCA default value 750 nm (N50: off) Biuret default off
Bradford default value 750 nm (N50: 350 nm) Lowry default value 405 nm
Note: It is recommended to use the default baseline for each assay. 5. Select
curve fit type: Options are linear regression, zero regression (forces the
straight line
through the origin) and 2nd order regression.
57
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 6. Select Unit
7. Add up to 20 Concentrations by pushing on the Add Concentration button.
Added concentrations can be deleted with Enter the concentrations of the
standards for the standard curve.
8. Select Replicates none, 2 or 3
58
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
9. Measure a blank and depending on the replicate selection, all required
concentrations. Absorbances of the replicates will be shown in the results
area and if replicates are selected a mean value for each standard. It is
possible to exclude single measurements from the curve calculation by
switching the toggle switch off. Note: After the first sample measurement the
standard curve can no longer be altered. Once a Standard curve is created or
loaded from a stored method it will be used for concentration calculations of
measured samples. It may be necessary to do a blank measurement.
10. Apply sample and press the sample button to initiate the sample
measurement. Note: Once the sample measurement is initiated it is no longer
possible to change the standard curve.
SAVING AND LOADING STANDARD CURVES
It is possible to save measured standard curves as a Stored Method. To save
the standard curve push the store method button and enter a method name,
select a folder and save with the Store button. Methods can be opened in the
Stored Methods menu on the homescreen. Opening a saved Protein Assay method
shows a message with the option to load or remeasure the standard curve.
CALCULATIONS
Protein concentration is determined using the standard curve by correlating
absorbance values of samples with known concentration to calculate the
concentration of the unknown sample. In order to maintain accuracy and
precision please ensure that the R2 value of the standard curve is 0.95 or
greater.
59
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
KINETICS
Available only for C40-Go, N50-Go needs upgrade for activation of Kinetics.
METHOD OVERVIEW
The Kinetics application is useful for: measuring initial rates of enzyme-
catalyzed reactions, performing progress-curve analysis for complete
reactions, calculating basic MichaelisMenten parameters for single-substrate
reactions, and measuring enzyme inhibition. Simple kinetics studies, where the
change in absorbance is followed as a function of time at a fixed wavelength,
can be readily performed with the NanoPhotometer® Go. The rate of a chemical
reaction can be measured using spectrophotometric methods by studying the
change in absorbance at a fixed wavelength as a function of time. These
changes in absorbance reflect corresponding changes in the concentration of
reactants or products as the reaction progresses. The rate of many chemical
reactions can be markedly accelerated by the presence of catalysts, which
remain chemically intact during the reaction. Catalysts in the case of
biochemical reactions are generally represented by enzymes, which are
specialized protein catalysts. However, a few examples of special reactions
catalyzed by RNA molecules also exist. Studying the kinetics of a reaction can
reveal important details of the catalytic mechanism involved in terms of
sequence steps, transition state of reactants or nature of enzyme inhibitors.
MEASUREMENT PROTOCOL
Note: If a kinetic is started from a control device via WiFi connection
(tablet or smartphone) set the auto lock of the tablet or smartphone to never.
Otherwise the kinetics will be interrupted when the smartphone or tablet is
locked, because of losing the WiFi connection.
1. Select the Kinetics icon on the home screen
2. For Cuvette Application (C40-Go): Select the path length depending on the
used cuvette. Options are: 0.5 mm, 1 mm, 2 mm, 5 mm and 10 mm
If it is desired to heat the sample to 37°C use the toggle switch to turn on
the cell holder heater. When the cuvette holder has reached 37°C the toggle
color changes to green. Note: Only available for cuvette applications
(C40-Go).
60
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
For NanoVolume Application (only with N50-Go Upgrade): Select the dilution
depending on the sample concentration.
Note: There is no automatic path length setting available in this method.
Select either a virtual dilution of 15 (path length 0.67 mm) or of 140 (path
length 0.07 mm). 3. Default wavelength () is 340 nm but can be changed in the
range of 200900 nm (N50: 200650 nm), depending on the application. 4. Time
settings: Enter the duration time in minutes over which measurements are to be
taken. Possible
range is 13000 minutes. Enter the interval time between measurements in
seconds. Possible interval times are 5-
3,600 seconds (N50: 103,600 sec.), depending on the duration time. Enter the
delay time in seconds before the first measurement is taken. Possible delay
time is between 03,600 seconds, depending on the duration time.
Note: A maximum of 500 samples is possible. Please consider this when choosing
the duration and interval time. 5. Insert cuvette with the reference sample
and select blank button to initiate the measurement. 6. Insert cuvette with
the sample and select the sample button to initiate the measurements. Once the
kinetic is started the Blank button turns to a Pause/Continue button and the
Sample button to a Stop button. Note: While the kinetics is running it is not
possible to change the parameters, save data or delete data. Change of
parameters is only possible before starting the kinetic readings. Save and
delete data is only available after the kinetic session is stopped. Note: Auto
print and cryo label print is not available in Kinetics.
61
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
CALCULATIONS
All absorbance values are normalized to a 10 mm path.
A0 An dA Slope Final A
R2
= absorbance of start value (10 mm path)
= absorbance of actual value time n (10 mm path)
= absorbance of actual value absorbance of start value
= linear regression fit of all actual measurement points
= absorbance of final value
=
R2
=
ni=1
(Oi -Ei)2 Ei
[Oi
=
observed
slope
value;
Ei
=
expected
slope
value]
62
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
OD600
Available only for C40-Go, N50-Go needs upgrade for activation of OD600.
METHOD OVERVIEW
The growth of bacteria in liquid culture media is commonly monitored by
measuring the optical density at 600 nm (OD600) in small samples taken from
the cultures. OD600 measurements are typically used to determine the stage of
growth of the bacterial culture, thereby ensuring that cells are harvested at
an optimum point that corresponds to an appropriate density of live cells.
Growth of bacterial cells typically progresses through a series of consecutive
phases including: lag, log, stationary and decline (see Figure 1 on page 64).
In general, cells should be harvested towards the end of the log phase using
the optical density of the samples to determine when this point has been
reached. Since optical density in the case of OD600 measurements results from
light scattering rather than light absorption, this value varies depending on
the type of bacterial cells in the culture in terms of size and shape. Cells
are routinely grown until the absorbance at 600 nm (known as OD 600) reaches
approximately 0.4 prior to induction or harvesting. A linear relationship
exists between cell number (density) and OD 600 up to an absorbance value of
0.6, approximately.
As mentioned above, for turbid samples such as cell cultures, the absorbance
measured is due to light scattering, and not the result of molecular
absorption. Since the extent of scattering is affected by the optics of the
system (distance between the cell holder and instrument exit slit,
monochromator optics, slit geometry, etc.), different spectrophotometer types
will tend to give different OD 600 readings for the same turbid sample.
Therefore, if results from different spectrophotometers are to be compared,
they must be normalized first using appropriate calibration curves. For more
information see Technical Note #8 OD 600 which can be downloaded on the Implen
webpage: www.implen-go.com/scientificpublications/
A calibration curve can be constructed by comparing measured OD 600 to
expected OD 600. Expected OD 600 is determined by counting cell number using
an alternative technique (for example microscope slide method) and converting
to OD 600 using the rule of thumb that 1 OD 600 = 5 x 108 cells/ml for E.
coli.
The NanoPhotometer® Go comes with a correction factor of 1 by default. To
compare OD 600 values between different spectrophotometers, it is necessary to
determine the constant deviation or ratio between the absorbance values for
the same sample from each instrument and use this factor within the setting
“correction factor” of your NanoPhotometer® Software.
Note: The use of 10 mm path length disposable cuvettes is recommended for
optical density measurements of cell culture solutions. The amount of cells is
reflected in the reading and the likelihood of fluctuating amount of cells in
a drop from sample to sample can be considered as extremely significant. It is
therefore recommended to use cuvettes since the amount of error in a bigger
volume is not as significant. The cuvette measurements provide a bigger
average and therefore more reproducible readings. Also, to prevent the
suspension settling too quickly and giving an OD reading that changes with
time, glycerol should be added to the sample.
63
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
Figure 1 Bacterial growth curve
MEASUREMENT PROTOCOL
1. Select the OD600 icon on the home screen
2. For Cuvette Application (C40-Go): Select the path length depending on the
used cuvette. Options are: 0.5 mm, 1 mm, 2 mm, 5 mm and 10 mm
If it is desired to heat the sample to 37°C use the toggle switch to turn on
cell holder heater. When the cuvette holder has reached 37°C the toggle color
changes to green. Note: Only available for cuvette applications (C40-Go). For
NanoVolume Application (only with N50-Go Upgrade): Select the dilution
depending on the sample concentration
Note: There is no automatic path length setting in this method. Select either
a virtual dilution of 15 (path length 0.67 mm) or of 140 (path length 0.07 mm)
3. Default wavelength is 600 nm but the wavelength can be changed in the
range of 200900 nm (N50-Go: 200650 nm), depending on the application.
4. Toggle switch cells/ml is disabled by default. Enable cells/ml to get the
cells/ml calculated. Enter the cell specific factor and multiplier (e.g. 1
OD600 = 5 x 108 cells/ml) 64
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
5. Enter the correction factor to compensate for different optical
configurations between the NanoPhotometer® Go and other instruments.
6. Option to smooth the graph with different boxcars. Options: Off, 1 =
boxcar 11 (default), 2 = boxcar 21 and 3 = boxcar 61
7. Option to set/calculate a dilution factor for manual diluted samples.
8. Insert cuvette with the reference sample and select blank button to
initiate the measurement 65
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
9. Insert cuvette with the sample and select the sample button to initiate
the measurement.
CALCULATIONS
OD600 = A600 Ð cf
OD600 A600 Ð cf
Optical density at 600 nm Absorbance at 600 nm (10 mm path) Dilution factor Correction factor for spectrophotometer
Cells/ml = A600 Ð cf * multiplier
A600 Ð cf multiplier
Absorbance at 600 nm (10 mm path) Dilution factor Correction factor for spectrophotometer Multiplier of sample
66
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
MORE APPS
Available only for C40-Go, N50-Go needs upgrade for activation of More Apps.
The More Apps icon located on the home screen opens another menu screen with
access to icons for additional applications available on the NanoPhotometer®
Go. The applications featured in this menu include: wavelength, concentration,
wavescan, absorbance/ratio, standard curve and custom applications.
MORE APPS: WAVELENGTH
Available only for C40-Go, N50-Go needs upgrade for activation of Wavelength.
METHOD OVERVIEW
In the wavelength application it is possible to measure simple absorbance (A)
and % transmittance (%Trans. / only in cuvette mode) of a sample at specific
wavelengths. It is possible to add up to 20 different wavelengths. The
wavelength method includes a calculation tool to define and calculate customer
defined formulas.
MEASUREMENT PROTOCOL
1. Select the More Apps icon from the home screen and the Wavelength icon
from the More Apps screen.
2. For cuvette application (C40-Go): Select the path length depending on the
used cuvette. Options are: 0.5 mm, 1 mm, 2 mm, 5 mm and 10 mm
If it is desired to heat the sample to 37°C use the toggle switch to turn on
cell holder heater. When the cuvette holder has reached 37°C the toggle color
changes to green. Note: Only available for cuvette applications (C40-Go). For
NanoVolume application (only with N50-Go Upgrade): Select the dilution
depending on the sample concentration.
67
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
Note: There is no automatic path length setting in this method. Select either
a virtual dilution of 15 (path length 0.67 mm) or of 140 (path length 0.07
mm). 3. Enter desired wavelength () to be measured. It is possible to measure
up to 20 wavelengths simultaneously. More wavelength () options can be added
by selecting the Add Wavelength button. Added wavelength can be deleted with
4. Baseline correction is set off by default. Enabling the baseline
corrections shows a list with different wavelength options: 377 nm, 604 nm,
650 nm, 770 nm (N/A N50-Go) and 823 nm (N/A N50-Go). Option to enter any
wavelength between 200 nm and 900 nm (N50-Go: 650 nm).
5. Option to smooth the graph with different boxcars. Options: Off, 1 =
boxcar 11 (default), 2 = boxcar 21 and 3 = boxcar 61
6. Option to enter formulas to calculate
68
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
It is possible to enter up to 5 different custom formulas for result
calculation. Formula Acceptance Guidelines: 1. Numbers:
Up to 20 significant figures if no decimal separator is used Up to 4
significant figures if a decimal separator (period “.”) is used 2. Numeric
Operations: + (add), – (subtract), (multiply), / (divide) and parentheses ()
3. Absorbance: Axxx e.g. for absorbance at 260 nm: A260 Note: Do not use blank
character. Example: Nucleic Acid (dsDNA) concentration calculation with
background correction at 320 nm: (A260-A320)50 7. Option to set/calculate a
dilution factor for manual diluted samples.
8. Apply the blank ddH20 or buffer to the illuminated sample window on
pedestal for the reference measurement and select blank to initiate the
reading.
Note: The illumination of the sample window can be switched off in
preferences.
69
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
9. Use a lint-free laboratory wipe to clean both the sample window on
pedestal and mirror in lid arm prior to applying the next sample. Note: It can
be helpful to apply the blank a second time and read it as a sample to ensure
a proper blank.
Apply sample to the sample window on pedestal and press the sample button to initiate the measurement.
CALCULATIONS
Formula Calculation: Is depending on the entered formula in the parameter
concentration.
Absorbance Calculation:
Absorbance is formally defined as the decimal logarithm (base 10) of the
reciprocal of transmittance:
A = log (T1) = – log T T = 10(-A)
Note: Corresponding absorbance value e.g. Absorbance value ( = 230) etc. normalized to 10 mm path length
%Transmittance Calculations (cuvette mode only)
In the wavelength application it is possible to measure the absorbance (A) and % transmittance (%T) of a sample with respect to a reference at a specific wavelength. Transmittance is the ratio of light intensity remaining after it has passed through the sample (I) to the initial incident light intensity (I0):
=
0
%
=
0
×
100
70
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
MORE APPS: WAVESCAN
Available only for C40-Go, N50-Go needs upgrade for activation of Wavescan.
METHOD OVERVIEW
Using the wavescan application it is possible to obtain the complete spectral
scan for a defined wavelength range between 200-900 nm (C40-Go) or from
200-650 nm (N50Go/Upgrade necessary).
MEASUREMENT PROTOCOL
1. Select the More Apps icon from the Home screen and the Wavescan icon from
the More Apps screen
2. For cuvette application (C40-Go): Select the path length depending on the
used cuvette. Options are: 0.5 mm, 1 mm, 2 mm, 5 mm and 10 mm
If it is desired to heat the sample to 37°C use the toggle switch to turn on
cell holder heater. When the cuvette holder has reached 37°C the toggle color
changes to green. Note: Only available for cuvette applications (C40-Go). For
NanoVolume application (only with N50-Go Upgrade): Select the dilution
depending on the sample concentration Note: There is no automatic path length
setting in this method. Select either a virtual dilution of 15 (path length
0.67 mm) or of 140 (path length 0.07 mm)
3. Set Start and End Wavelength to define the scan range.
Note: If samples with a different wavelength range are selected, the graphs
are shown on full scan range of 200900 nm (N50-Go: 200-650 nm). 4. Baseline
correction is set off by default. Enabling the baseline corrections shows a
list with
71
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 different wavelength
options: 377 nm, 604 nm, 650 nm, 770 nm (N/A N50) and 823 nm (N/A N50). Option
to enter any wavelength between 200 nm and 900 nm (N50: 650 nm)
5. Option to smooth the graph with different boxcars. Options: Off, 1 =
boxcar 11 (default), 2 = boxcar 21 and 3 = boxcar 61
6. Option to set/calculate a dilution factor for manual diluted samples.
7. Apply the blank ddH20 or buffer to the illuminated sample window on
pedestal for the reference measurement and select blank to initiate the
reading. Note: The illumination of the sample window can be switched off in
the preferences.
8. Use a lint-free laboratory wipe to clean both the sample window on
pedestal and mirror in lid arm prior to applying the next sample. Note: It can
be helpful to apply the blank a second time and read it as a sample to ensure
a proper blank.
72
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 9. Apply sample to the
sample window on pedestal and press the sample
button to initiate the measurement.
CALCULATIONS
No calculations necessary: values are reported based on 10 mm path length. The
results show prominent peaks with wavelength and absorbance values. For
cuvette measurements it is possible to change to the %Transmittance mode. If a
peak of interest is not shown in the results the peak can be selected by
pushing on the graph. The peak can then be added to the results by tapping on
the Add Peak button in the pop-up window.
73
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
MORE APPS: ABSORBANCE RATIO
Available only for C40-Go, N50-Go needs upgrade for activation of Absorbance
Ratio.
METHOD OVERVIEW
In this mode, it is possible to determine simple absorbance ratios for a given
sample by measuring the absorbance at two wavelengths specified in the
parameters of the method relative to a blank.
MEASUREMENT PROTOCOL
1. Select the More Apps icon from the home screen and the Absorbance/Ratio
icon from the More Apps screen
2. For cuvette application (C40-Go): Select the path length depending on the
used cuvette. Options are: 0.5 mm, 1 mm, 2 mm, 5 mm and 10 mm
If it is desired to heat the sample to 37°C use the toggle switch to turn on
cell holder heater. When the cuvette holder has reached 37°C the toggle color
changes to green. Note: Only available for cuvette applications (C40-Go). For
NanoVolume application (only with N50-Touch Upgrade): Select the dilution
depending on the sample concentration
Note: There is no automatic path length setting in this method. Select either
a virtual dilution of 15 (path length 0.67 mm) or of 140 (path length 0.07
mm). 3. Enter desired wavelengths ( 1-1 and 1-2) for ratio calculation. It is
possible to measure up to 20 absorbance/ratios simultaneously. More
wavelengths for ratio calculation can be added by selecting the Add Ratio
button. Added ratios can be deleted with .
74
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 4. Baseline correction
is set to off as default. Enabling the baseline corrections shows a list
with different wavelength options: 377 nm, 604 nm, 650 nm, 770 nm and 823 nm.
Option to enter any wavelength between 200 nm and 900 nm (N50: 650 nm)
5. Option to smooth the graph with different boxcars. Options: Off, 1 =
boxcar 11 (default), 2 = boxcar 21 and 3 = boxcar 61
6. Option to set/calculate a dilution factor for manual diluted samples.
7. Apply the blank ddH20 or buffer to the illuminated sample window on
pedestal for the reference measurement and select blank to initiate the
reading. Note: The illumination of the sample window can be switched off in
the preferences.
8. Use a lint-free laboratory wipe to clean both the sample window on
pedestal and mirror in lid arm prior to applying the next sample. Note: It can
be helpful to apply the blank a second time and read it as a sample to ensure
a proper blank.
75
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
9. Apply sample to the sample window on pedestal and press the sample button
to initiate the measurement.
CALCULATIONS
The absorbance ratio is calculated from the two path lengths specified by the user in the parameters.
1: 2=
1 2
1: 2 = Absorbance Ratio 1 = Absorbance 1 corresponding absorbance value 1
selected normalized to 10 mm path
2 = Absorbance 2 corresponding absorbance value 2 selected normalized to 10 mm
path
76
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
MORE APPS: CONCENTRATION
Available only for C40-Go, N50-Go needs upgrade for activation of
Concentration.
METHOD OVERVIEW
In this mode, concentration can be calculated for a sample by determining the
absorbance at a specific wavelength relative to a reference. The concentration
is then obtained by multiplying the measured absorbance by a specific factor.
This factor may be known in advance and entered by the user, or it may be
calculated by the instrument by measuring a set of standard (standard curve
method) with known concentrations to create a standard curve.
MEASUREMENT PROTOCOL
1. Select the More Apps icon from the home screen and the Concentration icon
from the More Apps screen.
2. For cuvette application (C40-Go): Select the path length depending on the
used cuvette. Options are: 0.5 mm, 1 mm, 2 mm, 5 mm and 10 mm
If it is desired to heat the sample to 37°C use the toggle switch to turn on
cell holder heater. When the cuvette holder has reached 37°C the toggle color
changes to green. Note: Only available for cuvette applications (C40-Go). For
NanoVolume application (only with N50-Go Upgrade): Select the dilution
depending on the sample concentration.
Note: There is no automatic path length setting in this method. Select either
a virtual dilution of 15 (path length 0.67 mm) or of 140 (path length 0.07
mm). 3. Default wavelength is 260 nm but can be changed in the range of 200900
nm (N50: 200650 nm), depending on the sample/application. 4. Enter a factor
for concentration calculation.
77
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 5. Unit selection
6. Baseline correction is set off by default. Enabling the baseline
corrections shows a list with different wavelength options: 377 nm, 604 nm,
650 nm, 770 nm (N/A N50) and 823 nm (N/A N50). Option to enter any wavelength
between 200 nm and 900 nm (N50: 650 nm)
7. Option to smooth the graph with different boxcars. Options: Off, 1 =
boxcar 11 (default), 2 = boxcar 21 and 3 = boxcar 61
78
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
8. Option to set/calculate a dilution factor for manual diluted samples.
9. Apply the blank ddH20 or buffer to the illuminated sample window on
pedestal for the reference measurement and select blank to initiate the
reading. Note: The illumination of the sample window can be switched off in
the preferences.
10. Use a lint-free laboratory wipe to clean both the sample window on
pedestal and mirror in lid arm prior to applying the next sample. Note: It can
be helpful to apply the blank a second time and read it as a sample to ensure
a proper blank.
11. Apply sample to the sample window on pedestal and press the sample button
to initiate the measurement.
79
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
CALCULATIONS
In this method, the concentration of the sample is calculated based on the
Beer-Lambert law given the user specified wavelength of interest and user
defined extinction coefficient. The equations for calculating concentration
without background correction are as follows:
Without background correction:
= Ð
C
Concentration (ng/µl)
Absorbance at user specified path length n (10 mm path)
Ð
Dilution factor
extinction coefficient/factor
80
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
MORE APPS: STANDARD CURVE
Available only for C40-Go, N50-Go needs upgrade for activation of Standard
Curve.
METHOD OVERVIEW
The construction of a calibration curve from multiple standards of known
concentrations can be created and stored on the NanoPhotometer® Go. The
standard curve can be used to quantify samples of the same type with unknown
concentrations. This application provides an extremely useful tool with which
to integrate, expedite and simplify the measurement and calculations involved
in determining the concentration of analytes in unknown samples. If a zero
concentration standard is required, include it in the number of standards to
be entered using a reagent blank and entering 0.00 for concentration.
MEASUREMENT PROTOCOL
1. Select the More Apps icon from the home screen and the Standard Curve icon
from the More Apps screen.
2. For cuvette application (C40-Go): Select the path length depending on the
used cuvette.
If it is desired to heat the sample to 37°C use the toggle switch to turn on
cell holder heater. When the cuvette holder has reached 37°C the toggle color
changes to green. Note: Only available for cuvette applications (and C40-Go).
3. For NanoVolume application (only with N50-Go upgrade): Select the dilution
depending on the sample concentration
Note: There is no automatic path length setting in this method. Select either
a virtual dilution of 15 (path length 0.67 mm) or of 140 (path length 0.07
mm). 4. Baseline correction is set off by default. Enabling the baseline
corrections shows a list with different wavelength options: 377 nm, 604 nm,
650 nm, 770 nm (N/A N50) and 823 nm (N/A N50). Option to enter any wavelength
between 200 nm and 900 nm (N50: 650 nm)
81
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 5. Select the curve
fit type: Options are linear regression, zero regression (forces the straight
line through the origin) and 2nd order regression. 6. Select Unit
82
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 7. Add up to 20
Concentrations by pushing on the Add Concentration button. Added
concentrations can be deleted with Enter the concentrations of the standard
curve.
8. Select Replicates none, 2 or 3
9. Once Standard curve is created or loaded it will be used for concentration
calculations in the method. It might be necessary to do a blank measurement.
10. Apply sample and press the sample button to initiate the measurement.
Note: Once the sample measurement is initiated it is not possible to make
changes to the standard curve.
SAVING AND LOADING STANDARD CURVES
It is possible to save measured standard curves as a Stored Method. To save
the standard curve push the store method button and enter a method name,
select a folder and save with the Store button. Methods can be opened in the
Stored Methods menu on the homescreen. Opening a saved Protein Assay method
shows a message with the option to load or remeasure the standard curve.
83
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
CALCULATIONS
Concentration is determined via the absorbance values provided by the standard
curve based on the curve fit selection including the following options: linear
regression, zero regression and 2nd order regression.
84
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
CUSTOM APPS
There is an option for designing customer specific Custom Apps which can be
loaded to the NanoPhotometer® Go. For more information about designing custom
applications to suit individual research needs please contact Implen directly
for assistance.
85
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
STORED RESULTS
The Stored Results icon opens a directory of folders containing files of
results that have been previously saved.
On the left side of the screen all available directories/storages are shown:
NanoPhotometer®, Control Device, Network and/or USB flash drive (depending on
availability). Pushing a storage folder shows the subfolders of this storage
folder on the left side and the individual files on the right side. On the
right side of the screen all saved result files of the selected folder are
shown and can be opened by a long or double click. Folders can be deleted,
renamed, moved or copied by pushing on the icon. It is also possible to
delete, rename, move or copy files by pushing on the icon. The file path of
the selected folder is shown on the top of the right file area. Note: PDF and
Excel files cannot be opened on the NanoPhotometer® Go. Files need to be
transferred to a computer or device where Excel or a PDF reader is installed.
Note: Control device is only available on computer, tablets and smartphones
not on the NanoPhotometer® Go version of the software. For data transfer via
Ethernet or WiFi see page 38 Data Transfer. Backup copies are saved in the
Autosave folder of the NanoPhotometer® Go (Stored
Results/NanoPhotometer/Autosave) for up to ten days. After ten days the
autosave files are automatically moved to an autosave archive folder. The
autosave archive folder can only be accessed via NanoPhotometer® Go file
server. Data in the autosave archive folder are not automatically deleted. The
content of the Autosave Archive folder can be deleted via the action button in
Stored Results. Make sure that a backup is created before deleting the
Autosave Archive folder content.
86
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
STORED METHODS
The stored methods icon opens the directories of folders containing methods
stored by the user.
On the left side of the screen all available directories/storages are shown:
NanoPhotometer®, Control Device, Network and/or USB flash drive (depending on
availability). Pushing a storage folder shows the subfolders of this method
folder on the left side and the individual files on the right side. On the
right side of the screen all saved method files of the selected folder are
shown and can be opened by a long or double click. On the top of the right
area the file path of the selected folder is shown. New folders can be created
by pushing on . Folders can be deleted, renamed, moved or copied by pushing on
the icon. It is also possible to delete, rename, move or copy methods by
pushing on the icon.
87
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
5. PREFERENCES
System preferences can be set by selecting preferences on the home screen. The
preferences menu includes: General, Dyes, Warning Messages, Network, Printer
and CFR21. The preference options of the selected menu item are listed in the
window on the right. Note: Preferences are not available on smartphones with
screen sizes less than 7 inches.
GENERAL
Selecting General in the Preferences menu opens a window to the right of the
preferences menu with the following options: Date and Time, Display, About,
Storage and Illumination Sample Window (N50-Go).
DATE AND TIME
Within the Date and Time it is possible to set the actual date and time of the
NanoPhotometer® Go or change the time zone.
To change the time zone or the date and/or time, push on the appropriate field
to open the selection options. The changed settings are shown below the time
zone field. To apply the changes, the NanoPhotometer® Go must be rebooted.
Start the reboot by pushing on the Set and Reboot button.
88
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2 Note: Do not change
the date and time at the same time as the time zone. Make sure that the UTC
time is set correctly before changing the time zone.
DISPLAY
Brightness: adjustment of the built-in screen brightness
ABOUT
In About the following information of the NanoPhotometer® Go are shown:
NanoPhotometer® Version, Serial Number, Ethernet IP Address, WiFi IP Address,
Hardware Version, Firmware Version, Time & Date of Initialization Test and
Status of Initialization.
STORAGE
Shows the total storage capacity and the free space of the internal
NanoPhotometer® Go storage.
ILLUMINATION SAMPLE WINDOW
Toggle switch to switch on/off the illumination of the sample window (for
N50-Go only)
DYES
There is a list of preprogrammed dye-labels for both nucleic acid dyes and
protein dyes. To toggle between the nucleic acid and protein list push on the
Nucleic Acid/Protein buttons in the header.
89
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
Each dye has either a lock icon ( ) in front of the dye name indicating that
the dye is locked and cannot be changed or a delete symbol ( ). The delete
option is only available for unlocked and not preprogrammed dyes. Selecting a
dye name opens a new screen with the dye information: dye name, absorbance
maximum dye (nm), dye-dependent extinction coefficient dye (M-1 cm-1), and
dyedependent correction factor as well as the option to show the dye in the
parameter list of the application (Nucleic Acid or Protein UV).
Note: It is not possible to delete a dye from the default factory list; custom
dyes can be deleted if they are not locked. It is possible to add a new dye to
the list by selecting the + button to add a new dye. A window will open where
it is possible to enter the: dye name, dye absorbance maximum (nm), dye-
dependent extinction coefficient dye (M-1 cm-1), and dye-dependent
correction factor. There is a toggle switch available to lock the dye to
prevent deleting a dye from the dye list accidentally.
90
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
WARNING MESSAGES
BLANK CONTROL
Toggle switch to switch on/off the Blank ControlTM of the NanoPhotometer® Go.
Note: Blank ControlTM is available for all NanoVolume methods (N50-Go).
SAMPLE QUALITY CONTROL
It is possible to change the upper and the lower limit of the ratio alert
warning messages. Default values for nucleic acid ratios are: 260/230 ratio
1.8 A – 3 A and 260/280 ratio 1.65 A – 2.5 A. Default value for Protein UV
ratio is: 260/280 ratio is 0.7 A.
91
NanoPhotometer® N50-Go/C40-Go User Manual Version 4.6.2
NETWORK
Selecting Network in the Preferences menu opens a window to the right of the
preferences menu with the following options: Network Settings, WLAN Settings,
File Server Access and Network Folder.
References
- Go.com | The Walt Disney CompanyÂ
- Implen-GO N50-GO, C40-GO, OD600-GO | Best in Spectrophotometers
- Implen-GO N50-GO, C40-GO, OD600-GO | Best in Spectrophotometers
- Downloads Spectrophotometer Manuals, Software
- Implen NanoPhotometer | UV-Vis Spectrophotometers | Downloads
- Implen NanoPhotometer | UV-Vis Spectrophotometers | Downloads