euromex BS.1152-PLPHi Bscope Phase Contrast Microscope Instruction Manual

BS.1152-PLPHi Bscope Phase Contrast Microscope

Instruction Manual

Introduction

Thank you for purchasing the Euromex bScope.

The bScope series has been designed with all kinds of Life Sciences applications and great durability in mind. This resulted in a modern, robust and high-level microscope for advanced use, equipped with the best optical and mechanical components. Specific attention to production methods resulted also in an excellent price/performance ratio

Please read this manual carefully before using this product to ensure correct and safe usage

• The content of this manual is subject to change without notice
• The appearance of the actual product can differ from the models described in this manual
• Not all equipment mentioned in this manual has to be part of the set you have purchased
• All optics are anti-fungus treated and anti-reflection coated for maximum light throughput

General safety instructions

Intended use: a non-medical device
This microscope is intended for general observation of cells and tissues, with transmitted/reflected illumination and with the specimen fixed on a slide

Intended use: medical device class I
This microscope is intended for observation and diagnostics of cells and tissues at hospitals or by physicians and veterinaries in private practice in pathology, anatomy and cytology applications. To be used with transmitted/ reflected illumination and with the specimen fixed on a slide. Physicians and veterinaries use microscopes to identify the different types of cells and spot abnormal cells. This product helps in identifying and treating diseases

Dangers associated with the operation

• Improper use could result in injury, malfunction or damage to property. It must be ensured that the operator informs every user of existing hazards
• Danger of electrocution. Disconnect the power to the entire lighting system before installing, adding or changing any component
• Not to be used in corrosive or explosive environments
• Avoid direct exposure of eyes to the collimated light beam or direct light from the light guides or fibres
• To avoid a hazard to children, account for all parts and keep all packing materials in a safe place

Photobiological safety LED, important safety instructions

• Avoid direct eye exposure to any LED light source while switched on
• Before looking through the eyepieces of the microscope, lower the intensity of the LED illumination
• Avoid long and high-intensity exposure to LED light because this may cause acute damage to the retina of the eye

Prevention of biological and infectious hazards
Infectious, bacterial or viral biohazard substances under observation may be a risk to the health of humans and other living organisms. Special precautions should be taken during in vitro medical procedures:

• Biological hazards: keep a logbook of all the biological substances or pathogenic microorganisms that were under observation with the microscope and show it to everybody before they use the microscope or before they do some maintenance work on the microscope! Agents can be bacterial, spores, enveloped or non­enveloped virus particles, fungi or protozoa
• Contamination hazard:
• A sample that is properly enclosed with a cover glass never comes in direct contact with the microscope parts. In that case prevention of contamination lies in the handling of the slides; as long as the slides are decontaminated before use and are undamaged and treated normally, there is virtually zero risk of contamination
• A sample that is mounted on a slide without cover glass, can come in contact with components of the microscope and may be a hazard to humans and/or the environment. Therefore, check the microscope and accessories on possible contaminations. Clean the microscope surfaces and its components as thoroughly as possible. Should you identify a possible contamination, inform the local responsible person in your organisation
• Microscope operators could be contaminated from other activities and cross-contaminate components of the microscope. Therefore, check the microscope and accessories on possible contaminations. Clean the microscope surfaces and its components as thoroughly as possible. Should you identify a possible contamination, inform the local responsible person in your organisation. it is recommended to wear sterile gloves when preparing the slides and handling the microscope in order to reduce contamination by the operator
• Infection hazard: direct contact with the focusing knobs, stage adjustments, stage and eyepieces/tubes of the microscope can be a potential source of bacterial and/or viral infections. The risk can be limited by using personal eyeshades or eyepieces. You can also use personal protections such as operation gloves and/or safety goggles, which should be changed frequently to minimize the risk
• Disinfectant hazards: before cleaning or disinfecting, check if the room is adequately ventilated. If not, wear respiratory protective gear. Exposure to chemicals and aerosols can harm human eyes, skin and respiratory system. Do not inhale vapours. During disinfection, do not eat, drink or smoke. Used disinfectants must be disposed of according to local or national regulations for health and safety

Disinfection and decontamination:

• Exterior casing and mechanical surfaces must be wiped with a clean cloth, dampened with a disinfectant
• Soft plastic parts and rubber surfaces can be cleaned by gently wiping a clean cloth, dampened with a disinfectant. Discoloration can occur if alcohol is used
• The front lens of eyepieces and objectives are sensitive to chemicals. We recommend not to use aggressive disinfectants but to use lens paper or a soft fibre-free tissue, damped in cleaning solution. Cotton swabs may also be used. We recommend you use personal eyepieces without eyes hades in order to minimize risk
• Never immerse or dip the eyepiece or objective into a disinfectant liquid! This will damage the component
• Never use abrasive compounds or cleaners that may damage and scratch optical coatings
• Properly clean and disinfect all possible contaminated surfaces of the microscope or contaminated accessories before storing for future use. Disinfection procedures must be effective and appropriate
• Leave the disinfectant on the surface for the required exposure time, as specified by the manufacturer. If the disinfectant evaporates before the full exposure time, reapply disinfectant on the surface
• For disinfection against bacteria, use a 70% aqueous solution of isopropanol (isopropyl alcohol) and apply for at least 30 seconds. Against viruses, we recommend to refer to specific alcohol or non-alcohol based disinfection products for laboratories

Before returning a microscope for repair or maintenance through a Euromex dealer, an RMA (return authorization form) together with a decontamination statement must be filled in! This document – available from Euromex for any reseller- must be shipped together with the microscope at all times

Reference documents:
World Health Organisation:
https://www.who.int/ihr/publications/biosafety-video-series/en/
Robert Koch lnstitut:
https://link.springer.com/content/pdf/10.1007/s00103-013-1863-6.pdf
US Centre for Disease Control and prevention
https://www.cdc.gov/infectioncontrol/guidelines/disinfection/index.html

Handle with care

• This product is a high quality optical instrument. Delicate handling is required
• Avoid subjecting it to sudden shocks and impacts
• Impacts, even small ones, can affect the precision of the instrument

Handling the LED
Note: Always disconnect the power cord from your microscope before handling the LED bulb and power unit and allow the system to cool down approximately 35 minutes to avoid burns

• Never touch the LED with your bare hands
•  Dirt or fingerprints will reduce the life span and can result in uneven illumination, lowering the optical performance
• Use only original Euromex replacement LEDs
• The use of other products may cause malfunctions and will void warranty
• During use of the microscope the power unit will get hot; never touch it while in operation and allow the system to cool down approximately 35 minutes to avoid burns

Dirt on the lenses

• Dirt on or inside the optical components, such as eyepieces, lenses, etc., affects the image quality of your system negatively
• Always try to prevent your microscope from getting dirty by using the dust cover, prevent leaving fingerprints on the lenses and clean the outer surface of the lens regularly
• Cleaning optical components is a delicate matter. Please, read the cleaning instructions further on in this manual

Model with rechargeable batteries

• Always disconnect the power cord from the microscope before you replace the rechargeable batteries
• The rechargeable batteries must not be thrown away as regular trash but should be taken to special waste collection sites, according your local or national regulations
• Risk of explosion: when removing the rechargeable batteries, do not throw the batteries into fire or any other heat source
• Do not replace the rechargeable batteries with non-rechargeable batteries
• Avoid extreme environmental conditions and temperatures which could affect the rechargeable batteries and lead to fire, explosion or leakage of hazardous substances
• If the rechargeable batteries have leaked, avoid contact of the chemicals with skin, eyes and mucous membranes
• When in contact with the chemicals, flush the affected areas immediately with plenty of fresh water and seek medical attention

Environment, storage and use

• This product is a precision instrument and it should be used in a proper environment for optimal use
• Install your product indoors on a stable, vibration free and level surface in order to prevent this instrument to fall thereby harming the operator
• Do not place the product in direct sunlight
• The ambient temperature should be between 5 to +40°C and humidity should be within 80% and 50%
• Although the system is anti-mold treated, installing this product in a hot, humid location may still result in the formation of mold or condensation on lenses, impairing performance or causing malfunctions
• Never turn the right and left focus knobs in opposite directions at the same time or turn the coarse focus knob past its farthest point as this will damage this product
• Never use undue force when turning the knobs
• Make sure that the microscope system can dissipate its heat (fire hazard)
• Keep the microscope away from walls and obstructions for at least approximately 15 cm
• Never turn the microscope on when the dust cover is in place or when items are placed on the microscope
• Keep flammable fluids, fabrick, etc. well out of the way

Disconnect power
Always disconnect your microscope from power before doing any maintenance, cleaning, assembling or replacing LEDs to prevent electric shocks

Prevent contact with water and other fluids
Never allow water or other fluids to come in contact with your microscope, this can cause short circuiting your device, causing malfunction and damage to your system

Moving and assembling

• This microscope is a relatively heavy system, consider this when moving and installing the system
• Always lift the microscope by holding the main body and base of the microscope
• Never lift or move the microscope by its focusing knobs, stage or head. When needed, move the microscope with two persons instead of one

Models
The bScope microscope is available in the following brightfield and phase contrast models.
Please note: on www.euromex.com you can find the latest updates about bScope models and accessories

bScope for brightfield

bScope for phase contrast

Objectives

The total magnification of the microscope can be calculated by multiplying the magnification of the eyepiece with the magnification of the objective. The magnifications are displayed in the table. The S40x, S60x and S100x objectives are equipped with a spring mount, to prevent damage to the front lens and the slide

Components of the microscope

The names of the several components parts are listed below and are indicated in the picture:

Preparing the bScope microscope for use
Carefully remove the items from its packaging and place them on a flat, firm surface. Please do not expose the microscope to direct sun light, high temperatures, damp, dust or acute shake.

Make sure the table or surface is flat and horizontal

When moving the microscope, use the left hand to hold the transport handle (A) and hold the base of the microscope with the right hand

Caution! Hold the microscope at the top of the stand arm when it is moved

Insert the power cord into the back of the microscope and use the cable storage system (CSS) to store excessive cable while in use or to store the cable after use (B)

Caution! If the bacterial solution or water splatters over the stage, objective or head, pull out the power cord immediately and dry the microscope

Assembling steps
Euromex Microscopen BV always tries to keep the number of assembly steps for their customers as low as possible but in

some cases there are some steps to be taken. The steps mentioned below are often not necessary but nonetheless described for your convenience nonetheless

Mounting the objectives

1. Rotate the coarse focusing knob to lower the stage to its lowest position
2. Install the objectives into the objective nosepiece from the lowest magnification to the highest in a clockwise direction from the rear side of the microscope. When using the microscope, start using the low magnification objective (4X or lOX) to search for specimen and focus, hereafter continue with high magnification objective to observe

The microscope head
The standard bScope series configuration is supplied with the head assembled. However, if your order contains a fluorescence or metallurgical attachment then this should be mounted first. There is a supplementary manual supplied with any intermediate attachment with detailed mounting instructions

Placing the eyepieces
The standard bScope series configuration is supplied with the eyepieces assembled. However, if your order contains additional eyepieces, please follow these steps to mount/replace them:

1. Remove the current eyepiece from the eyepiece tube, unlocking it with a screwdriver as shown in the pictures (A and B) on the next page

Note : the eyepiece tube with the diopter adjustment has three screws. In order to remove the eyepiece you need to unscrew the one aligned with ·o· (see picture C)

2. Insert the eyepiece into the eyepiece tube

3. lock the eyepieces with a screwdriver

The eyeshades (optional)
The eyepieces can be equipped with optional rubber eyeshades. This prevents damage to the lens, and prevents stray light. The eyeshade can simply be slipped over the eyepiece

Connecting the power cord
The bScope microscopes support a wide range of operating voltages: from 100 to 240V. Please use a grounded power connection

• Make sure the power switch is off before connecting
• Insert the connector of the power cord into the bScope power socket, and make sure it connects well
• Insert the other connector into the mains socket, and make sure it connects well. Do not bend or twist the power cord, it will get damaged. Use the power cord that is supplied by Euromex. If it is lost or damaged, choose one with the same specifications

Operation:

Setting up the illumination:
For optimal contrast and resolution one should follow the below procedure:

1. Place a specimen on the object stage and focus using the 4x objective, with a fully opened iris diaphragm
2. Turn light intensity to lowest position, then look through the eyepiece(s) and turn up to the comfortable light intensity level
3.  Turn the condenser in the highest position (for phase contrast models, please set condenser to brightfield position)
4.  Close the iris diaphragm, until it is just visible on the edge of the field of view

The microscope is properly set for use with the 4x objective. For each other magnification in brightfield use this procedure should be repeated to ensure the best balance between contrast and resolution. Phase contrast set up is explained later in this manual

STOP, Caution:
The maximum light intensity when using the 4x and 10x can damage the eyes!

Placing the specimen slide

1. Push the arm of the specimen holder backwards
2. Release the arm slowly clamping the slide with the cover glass facing up
3. Rotating the X and Y-axis knob will move the specimen to the center for alignment with the center of the objective

Focusing and slide protection

1. Select the 4x objective and make sure that it is placed correctly in the optical path
2. Move the stage to the top by using the coarse adjustment knob and focus till the image appears
3. Rotate the fine focusing knob to sharpen the image
4. When you perform focusing with a S100x objective, you need to lock the slide protection handle. The slide protection handle protects the slide by limiting the travel range of the mechanical stage. This way the objectives will not touch or damage your slides

1. The inter pupillary distance The inter pupillary distance
The correct inter pupillary distance is reached when one round image is seen in the field of view (see image below). This distance can be set by either pulling the tubes towards each other or pulling them away from each other. This distance is different for each observer and thus should be set individually. When more users are working with the microscope it is recommended to remember your inter pupillary distance for a quick set-up during new microscopy sessions. The bScope’s swiveling eyepiece tube can be rotated 360°.
You can select corresponding eye point height according to your own preference

2. The correct eye point
The eye point is the distance from the eyepiece to the user’s pupil. To obtain the correct eye point, move the eyes towards the eyepieces until a sharp image is reached at a full field of view

3. Adjusting the diopter

Using a binocular (or trinocular) tube is less tiring for the eyes than the use of a monocular tube. In order to obtain the right inter pupillary setting, one should go through the below steps

• Set diopter adjustment ring to zero
• Close the left eye and focus the right tube by adjusting the coarse-and fine adjustment knobs
• Close the right eye and focus the left tube with the diopter adjusting ring

This procedure be followed by each individual user. When more users are working on the same microscope it is recommended to remember your own diopter setting for a quick set-up during new microscopy sessions

Abbe condenser
Beneath the object stage an Abbe condenser N.A. 1.25 is mounted. The condenser can be adjusted in height by moving the rack and pinion knob beneath the mechanical stage. By adjusting the condenser you can focus the light on the specimen for a optimized contrast. The condenser is factory pre-centered. If needed, the following procedure can be followed to center the condenser

1. Move the condenser to its highest position
2. Select the 10x objective and place it into the light path and focus the specimen
3. Rotate the field diaphragm adjustment ring to put the field diaphragm to the smallest position
4. Adjust the condenser to the point where the image is the sharpest
5. Adjusting the center adjustment screw and put the image to the center of the field of view
6.  Open the field diaphragm gradually
7. The condenser is centered correctly if the image remains in the center when you open the field diaphragm and inscribed to the field of view

The field (Kohler) diaphragm (for Infinity models)
By limiting the diameter of the beam entering the condenser, the field diaphragm can prevent other light from entering and increase the image contrast. When the image is just on the edge of the field of view, the objective will show the best performance and obtain the clearest image. (see A in picture below.) The diaphragm is factory pre­centered

Adjusting the aperture diaphragm (see Bin picture below)

• The aperture diaphragm is used to select the numerical aperture (N.A.) of the illumination. When the N.A. of illumination matches with the N.A. of the objective, you get the highest possible resolution, depth of field and contrastare obtained
• When contrast is low, rotate the diaphragm adjustment ring to 70%-80% of the N.A. of objective this will improve the contrast of the image. The diaphragm is factory pre-centered

Use of the S1 00x oil-immersion objective
The Euromex bScope microscopes are equipped with an S100x N.A. 1.25 oil immersion objective. Please follow the below instructions on how to use this objective:

1. Remove the dust protection cap from the revolving nose piece to mount the SlOOx objective
2. Focus the image with the S40x objective
3. Lock the slide protection handle
4. Turn the revolving nosepiece so the S100x objective almost reaches the click-stop
5. Put a small drop of immersion oil on the center of the slide (always use Euromex Immersion oil)
6. Now turn the S 100x objective so that you feel the click stop
7. The front lens is in contact with the immersion oil
8. look through the eyepiece and focus the image with the fine adjustment knobs
9. The distance between the lens of the objective and the slide is very small!
10. In case there are small bubbles visible, turn the SlOOx objective a couple of times from left to right so that the front of the objective moves in the oil and the bubbles will disappear
11. After using the SlOOx objective, loosen the slide protection handle and turn the table with the course adjustment knobs downwards until the front lens doesn’t touch the oil any longer. Clean the front lens of the S100x objective
12. Always clean the front lens of the S100xobjective with a piece of lens paper that is moistened with a drop of isopropanol. We recommend using Euromex lens paper and isopropanol
13. Clean the slide after use as well

Illumination bScope series
The illumination has the following specifications:

LED: 3W NeoLED for binocular and trinocular models. .
Power supply: Primary AC 100 -240 Volt-SO Hz

Fuse specification: 250V 1A

Phase contrast

Use of phase contrast with the bScope microscope
The phase contrast method was designed in 1934 by the Dutchman Frits Zernike to observe very thin or transparent objects. This technique uses the fact that light travelling through tissue undergoes a phase shift due to diffraction
By recombining the phase shifted light with the background light, a contrasted image appears in the eyepiece

Using the Zernike phase contrast set
Any bScope model with a Zernike phase contrast set comes with the phase contrast condenser and objectives already mounted and centered on your microscope. If you suspect misalignment or want to check the alignment please see the next point for “centering the phase rings•
The height of the condenser can be adjusted by turning the rack and pinion up and down. By doing this the light beam will be focused more on the specimen for maximum resolution

Phase Contrast­ Principle

Centering the phase rings
The Zernike phase disc has five positions:
“0” for brightfield observation, this position also has an iris diaphragm
“10”
“20”
40″
“100”

These positions correspond to the respective phase contrast objectives 10x, 20x, 40x and 100x.
When the condenser is in the •o• position, the objectives can be used for brightfield observation. For phase contrast, the condenser position should match the objective used, meaning that when the condenser is in position •40• the objective used should also be 40x

• Rotate the 1 Ox infinity phase contrast objective into the field of view, then set the Zernike phase disc to position
  · 10· to match the objective

• Take the eyepiece out of the tube and insert centering telescope in its place. When looking through the centering telescope, the dark and bright ring images should coincide with each other as shown in the pictures below. If the ring images cannot be observed clearly, focus the centering telescope first (A) and if this does not solve the issue then try to adjust the condenser by turning it up and down (B).

• If the bright ring and dark ring images do not coincide as shown below, adjust the position of the ring by moving the ring at the bottom of the condenser with the centering knobs (C). Move it until the bright and dark ring images superimpose. Repeat for all objectives/Zernike disc positions

Using the phase contrast slider condenser (optional)

1. Keep the phase contrast slider facing up (text up); insert it from left to right into the condenser slider socket as the direction of the arrow indicates
2.  Each slider has three positions, two phase contrast positions and in the center of the slide the bright field brightfield position for normal use without phase contrast. Each phase contrast objective used has to be matched with the phase contrast ring on the slider.
   For example: when the 10x phase contrast objective is used, the slider should be positioneds to match the 10 phase diaphragm)

Note: the phase diaphragms in the sliders are pre­centered. It is not needed to adjust it before using

Maintenance and cleaning

Always place the dust cover over your bScope microscope
after use. Always keep the eyepiece and objectives mounted on the microscope to avoid dust entering the instrument

Cleaning the optics
When the eyepiece lens or front lens of the 10xor S40x objective are dirty, they can be cleaned by wiping a piece of lens paper over the surface (circular movements). When this does not help put a drop of alcohol on the lens paper and wipe it. Never put xylol or alcohol directly on the lens! Please note that Euromex offers a special microscope cleaning kit: PB.5275
It is not necessary – and not recommended – to clean the lens surfaces at the inner side of the objectives. Sometimes dust can be removed with high pressured air. There will never be dust in the objectives if the objectives are not removed from the revolving nosepiece
ill-; Caution Cleaning cloths containing plastic fibers can damage the coating of the lenses!

Maintenance of the stand
Dust can be removed with a brush. In case the stand or table is really dirty then you can clean the surface with a non­aggressive cleaning product. All moving parts like the height adjustment or the coaxial course and fine adjustment contain ball bearings that are not dust sensitive. With a drop of sewing-machine oil you can lubricate the bearingcan be lubricated

Replacing the fuse
To change the fuse, please follow these procedures below:

• Remove the power cord from the back of the microscope
• Locate the fuse compartment, which has a fuse image. It is typically located beneath the power connector
• Remove the fuse compartment. To do so, insert a flat head screw driver in between the metal power tines and gently pry the fuse compartment loose with a slight downward and outward motion
• Insert the new fuse into the compartment, and then replace the fuse compartment cover to where it originally was
• Power up the microscope and test

Note: The fuse may blow.in order to protect internal damage to the microscope. And iln most cases, replacing the fuse with the correct voltage will resolve the issue. However, should you encounter a blown fuse frequently, please contact your distributor for further assistance
Fuse specification: 250V, 3A

Replacing/placing the rechargeable batteries (optional)

• Remove the power cord from the back of the microscope

• Place the microscope on its back

• Remove the six screws of the base of the microscope
  Location of screws are indicated on drawing aside

• The battery compartment in located on the baseplate

• Open battery compartment, by removing the small screw on top, slide compartment open

• Place batteries and close the compartment

Note: Always use high quality rechargeable batteries, preferably supplied by Euromex. Minimum 1800mA capacity

type. Charge batteries fully for 8 hours. Use the microscope untill batteries are fully depleted then recharge again. Average use with full batteries: 8-32 hours, depending on light intensity and battery capacity and quality

Green battery Indicator: batteries are charged

Red battery indicator: batteries are being charged

Using the Kensington security slot
At the backside of the microscope a Kensington security slot is placed, which can be used to secure the instrument from theft using a Kensington lock (not supplied)

Digital cameras

Digital models are equipped with a build in digital camera inside. Connect the suppled USB cable to camera and follow the dedicated software manual for use. The LED which is placed beside the USB port will start to blink when activated by the software

Digital cameras are designed to be used on the photo port of the microscope head. It is also possible to use the digital camera in combination with a binocular head.

For To use the camera on a binocular bScopes, you can simple remove the eyepiece[1] and then place the camera with mounted c-mount adapter into the eyepiece tube[2]. Focus the digital image with the coarse and fine controls of the microscope

For trinocular models, slide the camera with mounted c-mount adapter into the 23,2mm tube of the photo port. Take an easy-to-view specimen and focus the image through the microscope’s eyepieces. For focusing the camera, slowly move tube (A) up and down while watching at the screen untill the camera view is in focus

Follow the manual that comes with the camera for camera operation

Accessories and spare parts

For current accessories and spares, please visit our website www.euromex.com
Euromex Microscopen by,  Papenkamp 20, 6836 BD Arnhem, The Netherlands
T +31 (0) 26 323 22 11, [email protected], www.euromex.com

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