IMPLEN OD600 DiluPhotometer User Manual

: June 6, 2024
: IMPLEN

IMPLEN OD600 DiluPhotometer™ User Manual

OD600

phone: + 49 (0) 89 726 3718 0 Fax. + 49 (0) 89 726 3718 54 Email: [email protected] www.implen.de

Declaration of Conformity

This is to certify that the Implen OD600 DiluPhotometer™ conforms to the requirements of the following directives:

2014/35/EU Low Voltage directive (LVD)
2014/30/EU Electromagnetic Compatibility directive (EMC)
2011/65/EU Restriction on the use of certain hazardous substances directive (RoHS)
2012/19/EU Waste electrical and electronic equipment directive (WEEE)
2006/42/EC Machinery directive

Standards used to demonstrate conformity include:

EN 61010-1:2010 Safety requirements for electrical equipment for measurement, control and laboratory use, General Requirements

EN 61010-2-101:2002 Safety requirements for electrical equipment for measurement, control and laboratory use.

EN 61326-1:2013 Electrical equipment for measurement, control and laboratory use –EMC requirements (class B)

EN ISO 12100:2010 Safety of machinery-General principles for design, risk assessment and risk reduction

Signed: Dated: October 18, 2021

Dr. Thomas Sahiri
Managing Director
Implen GmbH

ESSENTIAL SAFETY NOTES

Unpacking, Positioning and Installation

Check the contents of the pack against the packing list. If any shortages are discovered, please inform your supplier immediately.
Inspect the instrument for any signs of damage caused in transit. If any damage is discovered, please inform your supplier immediately.
Ensure your proposed installation site conforms to the environmental conditions for safe operation: Indoor use only Temperature 5°C to 35°C Maximum relative humidity of 80 % up to 31°C decreasing linearly to 50 % at 40°C
The instrument is powered by the internal rechargeable battery or by mains electricity using the supplied power-adapter. Using the instrument with the mains adapter will automatically recharge the battery.
The battery will last approx. 1 month when fully charged with normal use.
A full battery recharge will take approx. 12 hours (overnight).
Please read through this user manual prior to use.
Please contact your original supplier in the first instance if you experience technical or sample handling difficulties.

If this equipment is used in a manner not specified or in environmental conditions not appropriate for safe operation, the protection provided by the equipment may be impaired and instrument warranty withdrawn.

INTRODUCTION

Your Spectrophotometer

Your OD600 DiluPhotometer™ is a small, easy to use instrument that is dedicated to measuring samples at a wavelength of 600 nm. It is suitable for measuring growth rates of all types of cell (e.g. E. coli and yeast). Combining OD600 DiluPhotometer™ with DiluCell™ 10 facilitates measurements with low sample volumes of 300 µl and a virtual sample dilution of factor 10.

With a bandwidth of 40 nm the OD600 DiluPhotometer™ enables also measurements nearby 600 nm like the protein quantification assay according to Bradford (595 nm).

A 600 nm LED source in combination with a fibre optic is used to obtain the measurement. The instrument can be used in incubation cabinets and under anaerobic conditions.

Sample Handling Tips

Note that the light beam is directed horizontal from the front of the instrument to the back; therefore please ensure the cell is inserted in the correct alignment.
The optical height is 8.5 mm.
The cell holder supplied with the instrument accepts:

Sample Handling Tips

*Depends on tube being used.

IMPORTANT WARNING

Always wear protective clothing when handling bacteria or other cells.

Keypad and Display

DILUCELL™

Description

DiluCell™ is especially designed for the usage of the OD600 DiluPhotometer™ for the analysis of samples at a wavelength range of 340 nm to 950 nm. Due to the reduced pathlength, DiluCell™ provides a virtual dilution without the need of a physical dilution of higher concentrated samples.

DiluCell™ 10 allows an automatic 1/10 dilution of the sample. The required sample volume is 300 µl.

The following scheme explains the functionality of the DiluCell™ technology:

Lambert-Beer-Law:

Lambert-Beer-Law

Operation Instructions

Step 1

Place the DiluCell™ into the cuvette sample compartment. Ensure the correct positioning of the DiluCell™. The light beam of the OD600 DiluPhotometer™ is directed horizontal from the front of the instrument to the back.

Important Note: DiluCell™ will not work in any other orientation; do not mix other cuvette types within a DiluCell™ measurement series (for blank, etc.).

Step 2

Pipette your reference or sample into the DiluCell™ . DiluCell™ is very easy to fill; the geometrical setup supports air bubble free sample application. It is recommended however that you check for air bubbles before the measurement. In case of an air bubble in the cell please remove it by gently tapping the cell.

Volume requirement is: 300 µl for DiluCell™ 10

Important Information:

Remember to multiply the reading from the OD600 DiluPhotometer™ after the measurement with the proper virtual dilution factor:

Virtual dilution factor is: 10 for DiluCell™ 10

Important Note: It is not recommended to reuse DiluCell™ due to:

high probability of cross contamination
high probability of air bubbles when refilling the cell
decontamination not easily possible (for cell culture measurements, e.g.)

Specifications

Limitations

Do not autoclave DiluCell™.

The DiluCell™ is approved only for the usage with the OD600 DiluPhotometer™.

For solvent compatibility please contact your local Implen partner or the Implen Team at www.implen.de.

OD600 DILUPHOTOMETER™ – DILUCELL™ APPLICATION

Bacterial Cell Culture Measurement

Description

The stage of growth of a bacterial culture needs to be monitored to ensure that the cells are harvested at the optimum point for the greatest density of live cells. An exemplary growth curve is given below. Cells should be harvested towards the end of the log phase. The optical density of the sample indicates when this point has been reached. This value varies dependent on the cells being grown. Routinely the cells are 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 approx. 0.6.
It is important to note that for turbid samples such as cell cultures, the absorbance measured is due to light scattering, and not the result of molecular absorption. The amount of scatter is affected by the optics of the system (distance between the cell holder and instrument exit slit, geometry of this slit and the monochromator optics). Different spectrophotometer types therefore give different responses for the same turbid sample; to compare results, they must be normalized using calibration curves.
A calibration curve can be determined 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 = 8 x 108 cells/ml for E. coli. Clumping together of cells will also affect readings, so the medium they are suspended in will also make a difference.

Operation Instructions

Making a measurement:

Step 1 Switch the instrument on by pressing the ON/OFF button.
Step 2 Place a reference into the cuvette compartment.
Step 3 Press the blank button. The display will show 0.00.
Step 4 Remove the reference and replace with the sample solution in a cuvette.
Step 5 Press the sample button. The display will show the OD of the sample in absorbance units.

Important Information:

If using DiluCell™ remember to multiply the reading from the OD600 DiluPhotometer™ after the measurement with the proper virtual dilution factor:

Virtual dilution factor is: 10 for DiluCell™ 10

Multiple samples can be compared with the same reference by placing different samples in the cuvette chamber and making measurements for each one. It is recommended to re-reference with the reference solution every 10 to 15 minutes to avoid any slow instrument drift. Please re-reference always if in doubt.

Using the memory function

The instrument can store up to 99 readings in the memory. The results can then be viewed, printed or downloaded at a later time. This enables readings to be taken at, for example, an incubator and downloaded to a PC in a different laboratory. The results remain in the memory even when the instrument is switched off.

Step 1 Switch the instrument on by pressing the ON/OFF button.
Step 2 Press mem button to display MEM (if not already displayed).
Step 3 Place a reference into the cuvette compartment.
Step 4 Press the blank button. The display will show 0.00 but the memory number will not change.
Step 5 Insert the sample and press the sample button. The result will be displayed and automatically stored and the Memory Number will increase by one.
Step 6 To retrieve the results press recall/print. This will print out all of the results held in the memory if the instrument is connected to a PC or printer and cause the memory number to flash. Repeated pressing of the button will display the results in the memory on the screen in reverse order scrolling back to the beginning.
Step 7 Press reset or mem to go back to the latest result.
Step 8 Pressing reset when the latest result in the memory is showing will cause the screen to flash “rSt and ?”.
Step 9 If no further action is taken the screen will revert to its normal state after 7 seconds. If reset is pressed again whilst the screen is flashing all of the memory positions will be cleared.

Important Note: The instrument can store up to 99 readings.

The hundredth reading will automatically overwrite the first reading.

OD600 DILUPHOTOMETER™ – STANDARD CUVETTE APPLICATION

Protein Quantification with Bradford Assay

Description

Bradford Assay is a rapid and accurate method used to determine the total protein concentration of a sample by quantifying the binding of a dye, Coomassie Brilliant Blue G-250, to an unknown protein. The assay is based on the observation that the absorbance maximum for an acidic solution of Coomassie Brilliant Blue G-250 shifts from 465 nm to 595 nm when binding to protein occurs. By comparing this binding to that of different, known concentrations of a standard protein, the concentration of the unknown protein can be calculated. Standard curves with bovine serum albumin (BSA) are given below as an example.

Description 1

Operation Instructions

Important Note:
Please follow the detailed protocols supplied with your Bradford assay kit to ensure accurate results.

Making a measurement:

Step 1 Switch the instrument on by pressing the ON/OFF button.
Step 2 Place a reference into the cuvette compartment.
Step 3 Press the blank button. The display will show 0.00.
Step 4 Remove the reference and replace with standard solutions.
Step 5 Press the sample button. The display will show the OD of the standard solutions in absorbance units.
Step 6 Create a standard curve.
Step 7 Measure the absorbance of sample solutions.
Step 8 Determine the protein concentration of unknown samples according to the standard curve.

TROUBLE SHOOTING NOTES

ACCESSORIES

DiluCell™ 10 DC 10
Adapter set for 10 and 12 mm tubes OD600-AT

MAINTENANCE

Cleaning and General Care of the Instrument

The instrument has no serviceable parts.
The instrument requires little maintenance, but the following are considered good practice:

Keep the instrument clean and dry. Wipe off any spilt liquids immediately. Clean with a slightly damp cloth; a non-abrasive water-based soap or detergent may be used. The instrument may be wiped.
Remove the cuvettes from the instrument when not in use.
Store in a cool place away from corrosive chemicals or fumes.

De-contamination Procedure

To decontaminate we recommend that the instrument is wiped with ethanol or other antibacterial detergent as required. A soaked cloth may be inserted into the cuvette chamber or ethanol sprayed directly into the compartment.

The instrument can be sterilised using formaldehyde or ethylene oxide, but not with UV light (due to plastic degradation).

For severe contamination it is possible to remove the 4 screws in the base and separate the top and bottom covers (taking care to not drop the battery inside the instrument). The contaminated areas in the instrument may then be wiped with a suitable anti-bacterial detergent.

SPECIFICATION AND WARRANTY

Technical Specification

Technical Specification

Specifications are measured after the instrument has warmed up at a constant ambient temperature and are typical of a production unit. As part of our policy of continuous development, we reserve the right to alter specifications without notice. The product does not fulfil the specific requirements of the IVD.

Warranty

IMPLEN guarantees that the product supplied has been thoroughly tested to ensure that it meets its published specification. The warranty is as defined in our current terms and conditions and is only valid if the product has been used according to the instructions supplied. IMPLEN or your supplier can accept no liability for loss or damage, however caused, arising from the faulty or incorrect use of this product.