euromex OX.2253-PLF Oxion Inverso LifeScience Phase Contrast Microscope User Manual

euromix OX.2253-PLF Oxion Inverso LifeScience Phase Contrast

Microscope

Introduction

With your purchase of an inverted microscope of the Oxion Inverso range you have chosen for a quality product. The Oxion Inverso microscopes are developed for use at universities and laboratories The maintenance requirement are limited if the microscope is used in a decent manner
This manual describes the construction of the microscope, how to use the microscope (with optional phase contrast and ICSI equipment) and maintenance of the microscope

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 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

Phoobiological 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 nd 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 eyeshades 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.co m/content/pdf/10.100 7 /sOO 103-013-1863-6.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

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

Oxion Inverso available models

The microscopes of the Oxion Inverso series are available in two main configurations: two bright field models and two-phase contrast models. Additionally, the phase-contrast models can be supplied with complete IC51 equipment. See the table below for more details.

Construction of the microscope

The names of the several parts are listed below and are indicated in the picture (on the next page):

Assembling the microscope
Your microscope is pre-assembled at the factory. Some parts are not pre- assembled to prevent damage during transport:

• Eyepieces
• Phase slider (only in phase contrast models)
• Stage plate/s
• Mechanical stage

Mounting of the parts
Before mounting on the parts, make sure that the microscope is switched off and remove the main plug

Eyepieces
Remove the covers of the eyepiece tubes and insert the pair of eyepieces

Phase slider
The phase annuli slider (H) can be slid into the slot The phase alignment is already done by Euromex. Realigning the phase rings is only needed when you remove and change the objectives

Filter slider (optional)
The filters can be placed into the slider, which then can be slid into the slot (Kl

Stage plate/s
There are two types of stage plates supplied with the microscope. Take one and place it on the middle part of the stage

Mechanical stage
The mechanical stage should be mounted using two screws that are supplied with the mechanical stage (see picture)

General functions of the microscope

The stand consists of a stand arm, stand base, and an object stage. Hold the microscope at the top of the stand arm when it should be moved

Tubes
The fixed tubes are equipped with a pair of DIN HWF plan l0x/22 mm eyepieces

Revolving nosepiece
The revolving nosepiece on ball bearings can be equipped with five objectives

Optical specifications of the Oxion Inverso range
The Oxion Inverso range inverted microscopes are standard equipped with two compensation high wide field HWFl0x eyepieces

Table
The total magnification of your microscope can be calculated by multiplying the magnification of the eyepieces with the magnification of the objective. Total magnifications are displayed in the table below: Object stage
These models are supplied with either a flat object 250 x 230 mm stage or a 250 x 230 mm stage with coaxial mechanical 120 x 78 mm X-Y stage

Coarse and fine adjustment
The coarse/fine focusing knobs (D) for the height adjustment of the object stage are mounted together on one axle (i.e. coaxial). On one of the coarse adjustment knobs there is a graduation with intervals of 0.0025 mm. This can be used to measure depths in a specimen\

Illumination Oxion Inverso range
Transmitted 5 W LED intensity adjustable with internal 100-240 V power supply. Fuse holder 3.15 A/ 250 V

Preparing the microscope for use

After you have mounted on the parts indicated under section 4.1, the microscope is ready to use. Connect the plug into into the mains supply and switch on the microscope. Sit down comfortably behind the microscope and take a relaxed position while viewing through the eyepieces

Working with the microscope

Please read the following instructions to achieve the best microscope observing results
Setting the interpupillary distance

The correct interpupillary distance is reached when one round image is seen in the field of view (see image on the right). This distance can be set by either pulling the tubes towards each other or pulling them from each other. This distance is different for each observer and thus should be set individually. When several users are working with the same microscope, it is recommended to remember your interpupillary distance for a quick set up during new microscopy sessions
Setting the diopter adjustment

In case there is a difference in vision between both your eyes, you can use the microscope diopter adjustment to adjust it accordingly

• Turn the diopter adjustment ring of the left eyepiece tube until the scale shows the same reading as on the indicator
• Close the right eye and focus the left tube by means of the coarse and fine adjustment knobs
• Close the left eye and focus the right tube with the diopter adjustment ring

This procedure should be followed by each individual user
Caution:
The maximum light intensity when using the 4x and l0x objectives can damage the eyes!

Photo and video
The models are equipped with a vertical photo tube on which a camera adapter can be attached. This adapter is sold seperately and is needed to connect your camera with the microscope By moving out the photo/video switch of the trinocular head, 80% of the light is directed to the photo tube. The image remains visible through both eyepieces, with a capacity of 20%
Using the Oxion Inverso phase contrast slider

1. Keep the phase contrast slider face up (text up) and insert it from left to right into the condenser slider holder The phase contrast slider has three positions: two phase contrast positions {left and middle) and one bright field position for normal use without phase contrast (right). Each
2. phase contrast objective needs to be used in combination with the corresponding ring on the slider. For example: when working with the 10x phase contrast objective, the phase contrast ring in use should be “4X l0X” {i.e., the one on the left)

When working with phase contrast, please set the aperture diaphragm open (see A in photo next page)

Note: The phase diaphragms in the sliders are pre-centered do not need to be adjusted in operation Components:

• Phase contrast slider with three positions: phase contrast 4x/10x, phase contrast 20x/40x and bright field
• Centering telescope
• Centration keys
• Position of centration screw for 4X/10x phase contrast ring Position of centration screw for
• 20X/40x phase contrast ring

To check and correct the alignment, take the following steps:

• To check the 40x phase contrast objective, set the slider in such a way that the corresponding phase ring is in place under the condenser
• Place the centring telescope into the eyepiece tube, and focus the phase ring of the objective by means of the adjustable eye lens
• Now focus the centring ring of the condenser by means of the coarse and fine adjustment knobs
• If not correctly centred, the position of the rings in the slider can be adjusted by turning the two screws “E”
• To turn the screws “E’ use centration keys “C. Centration keys can be placed through the holes in the microscope condenser lamp house, then into screws ‘E•; see pictures on next page
• Now turn both centration keys “C” to move the phase contrast ring until it’s centred as shown below

This procedure has to be done for each individual objective. For 4x and 10x objectives use screw holes “D”

caution
Avoid contact with your finger and optical parts during centring

Maintenance and cleaning

Always place the dust cover over the microscope after use. Keep the eyepiece and objectives always mounted on the microscope to avoid dust entering the instrument

Cleaning the optics
When the eyepiece lens or front lens of the objectives is dirty they can be cleaned by wiping a piece of lens paper over the surface {circular movements). If this does not help put a drop of xylol or alcohol on the lens paper. Never put xylol or alcohol directly on the lens!

When dirt is clearly visible in the field of view it resides on the lowest lens of the eyepiece. By using the Allen-key the eyepiece can be removed from the tube. Clean the outside of the lens. In case there is still dust visible please check if the dust is in the eyepiece by turning it. If this is the case remove the lowest lens carefully from the eyepiece and clean it

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 nose piece
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 the surface can be cleaned 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 the bearing can be lubricated

Changing the LED
Caution
Always remove the power cable from the mains supply before changing W thelED!
Remove the covers of the Allen screws and unscrew the LED unit. Take out the unit and slide the the replacement LED unit {OX.9985) in its place. Reattach the Allen screws and covers

Rep lacing the fuse
Unscrew the fuse holder on the back of the microscope, using a straight screwdriver. Insert a new fuse in the holder and return it into the back of the microscope

Tension control setting
Between the right-handed coarse adjustment knob and the microscope stand there is a ring for the tension control setting. By means of turning it clock- or counter clockwise the tension of the coarse adjustment knobs can be adjusted

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