- Frank`s Hospital Workshop
Contents
1. condenser The method allows elements in the field of view to be distinguished that would be invisible or poorly visible using other methods of lighting such as brightfield or phase contrast light This method is commonly used to evaluate slides on which the object and the background are the same colour such as red blood cells in vital blood or clear bacterial organisms in saline In darkfield microscopy objects appear in the form of glowing outlines on a black dark background The advantage of darkfield illumination is that details can be seen that are normally not resolved by the microscope s objective While the actual details cannot be seen the light reflected by them can A nice analogy here is that of dust in a room Normally in a well lit room very small dust particles cannot be seen However if the lights are turned out a beam of light from an acute angle makes these same particles visible Besides the optical advantages darkfield illumination is very beautiful and gives an almost science fiction like image What is the difference between darkfield and brightfield The difference between the two is the way light is projected onto the slide As a result elements on the slide not visible or poorly visible in bright light are visible with darkfield illumination 8 2 Brightfield This is the basic microscope configuration In brightfield microscopv the transparent or translucent specimen is either naturallv coloured or stained and app2. 600x Eyepiece WF 10x 18 amp P 16x 11 Interpupillary Distance 55 75mm Nosepiece Inward Quintuple 5 hole Objective Lens Semi Plan DIN Achromatic Objectives Anti Fungal 4x 0 1 10x 0 25 40x 0 65 100x 1 25 Condenser Movable N A 1 25 Abbe Condenser with Iris Diaphragm Filtering Filter Holder and Daylight Blue Filter Stage Double Layer Mechanical Stage 160x140mm 75x50mm Dual Slide Holder Focusing Coaxial Coarse and Fine Focus Controls Focus Stroke 23mm Gradation 2um Collector Koehler Illumination Illumination Halogen Bulb 6V20W Adjustable Light Intensity Diagnostics Worldwide 3 10 HumaScope Plus User Manual 4 STRUCTURE 4 1 Base The base Figure 1 supports the entire weight of the microscope It has four rubber feet to stabilise the instrument The base contains the lamp source and the Complete electronic circuit Ta Vama ge eee gt ig Figure 1 4 2 Siedentopf Head The Siedentopf type ergonomic head figure 2 can be inclined at angles from 10 to 30 and the eyepieces can be extended up to 40mm The Siedentopf head does not require compensating adjustment for different interpupillary distances This ensures an optimum eye level and comfortable viewing position regardless of the operator s physique In addition a digital eyepiece camera can be attached to the tubes Figure 2 4 3 Illumination System In order to provide sufficient brigh
3. HUMASCOPE PLUS User Manual i Puma Sage pi CE Cat No 16201 REVISION LIST OF THE MANUAL No DATE Rev REVISION DESCRIPTION 1 01 2007 01 First edition Diagnostics Worldwide 1 INTRODUCTION This manual is considered as a part of the instrument it has to be at the operator s hand as well as at the maintenance operator s availability For accurate installation use and maintenance please read the following instructions carefully In order to avoid instrument or personal damages carefully read the GENERAL SAFETY WARNINGS describing the suitable operating procedures In case of breakdowns or any troubles with the instrument apply to the local Technical Service 2 USER WARRANTY HUMAN warrants that instruments sold by one of its authorised representatives shall be free of any defect in material or workmanship provided that this warranty shall apply only to defects which become apparent within one year from the date of delivery of the new instrument to the purchaser The HUMAN representative shall replace or repair any defective item at no charge except for transportation expenses to the point of repair This warranty excludes the HUMAN representative from liability to replace any item considered as expendable in the course of normal usage e g lamps valves syringes glassware fuses diskettes tubing etc The HUMAN representative shall be relieved of any liability under this warranty if the pro
4. NOTICE APPLICATIONS SPECIFICATIONS STRUCTURE 4 1 Base 4 2 Siedentopf Tube 4 3 Illumination System 4 3 1 Koehler Illumination 4 4 Mechanical Stage Slide Holder 4 5 Ocular Objective and Nosepiece 4 6 Coarse and Fine Focussing Knobs 4 7 Condenser 4 8 Immersion Oil OPERATION CARE AND MAINTENANCE DISPOSAL MANAGEMENT CONCEPT MICROSCOPE GLOSSARY 8 1 Darkfield Microscopy 8 2 Brightfield Microscopy 8 3 Phase Contrast Microscopy 8 4 Fluorescence Microscopy 8 5 Lenses 8 6 Focus O o 0 o oo Y NN NN A DO US A A A U U W FP LA O O O 2 10 1 NOTICE Every effort has been made to avoid errors in this text and the accompanying diagrams however HUMAN GmbH assumes no responsibility for any errors which may appear in this publication It is the policy of HUMAN GmbH to improve products as new technologies and components become available Human GmbH therefore reserves the right to change specifications if necessary in the course of such improvements 2 APPLICATIONS The HumaScope Plus biological microscope is suited for use in medical and sanitary establishments laboratories institutes agricultural research networks colleges and universities for clinical microscope inspections It can also be used in research work such as biology bacteriology cytology histology and pharmaceutical chemistry 3 SPECIFICATIONS Observation Tube Compensation Free Siedentopf type Binocular Head 302 inclined Magnification 40x 1
5. ar power source have to be dismounted from electric electronic parts and disposed off in accordance with applicable local regulations 6 INSTRUMENT DISINFECTION Instruments or parts which may come in contact with biological samples patient specimens controls etc should be considered at least potentially infectious Before doing any servicing on the instrument it is very important to thoroughly disinfect all possibly contaminated parts Before the instrument is removed from the laboratory for disposal or servicing it must be decontaminated disinfected Decontamination disinfection should be performed by authorised welltrained personnel observing all necessary safety precautions Instruments to be returned have to be accompanied by a disinfection certificate completed by the responsible laboratory manager If a disinfection certificate is not supplied the returning laboratory will be responsible for charges resulting from non acceptance of the instrument by the servicing centre or from authority s interventions 7 NOTICE Every effort has been made to avoid errors in text and diagrams however HUMAN GmbH assumes no responsibility for any errors which may appear in this publication It is the policy of HUMAN GmbH to improve products as new techniques and components become available HJMAN GmbH therefore has to reserve the right to change specifications if necessary in the course of such improvements BR _ W NH GO N M VI CONTENTS
6. d for transmitted light brightfield illumination 1 Switch on the light source and make sure that light is passing through the field diaphragm at the base of the microscope stand It may help to place a piece of paper over the field stop to confirm this Place your specimen on the stage and turn the nosepiece which holds the objective lenses to the 10X or 20X lens Open the field diaphragm as far as possible Check if the specimen is illuminated It will help to place a piece of paper over the top of the specimen to see if light is getting through If you are using the brightfield condenser stop open the iris diaphragm or aperture diaphragm on the condenser turret which contains the stops for brightfield and phase etc to the maximum The front lens should be about 1 3 mm above the specimen Use the condenser focussing knobs to do this Bring the specimen into focus with the coarse and fine focussing knobs If the light is too bright reduce the light intensity using the light adjustment wheel When the specimen is in focus start to close the field diaphragm and also begin to carefully move the condenser up and down with the condenser focussing knobs Look for a sharp image of the edge of the field diaphragm When the edge of the field diaphragm silhouette is sharply defined centre it with the two knurled knobs that extend diagonally from the condenser Close down the field diaphragm most or all the way to get it centred properly Wh
7. duct is not used in accordance with the manufacturer s instructions altered in any way not specified by HUMAN not regularly maintained used with equipment not approved by HUMAN or used for purposes for which it was not designed HUMAN shall be relieved of any obligation under this warranty unless a completed installation warranty registration form is received by HUMAN within 15 days of installation of this product This warranty does not apply to damages incurred in shipment of goods Any damage so incurred shall be reported to the freight carrier for settlement or claim 3 INTENDED USE OF THE INSTRUMENT The instrument has to be used for the expected purposes and in perfect technical conditions by qualified personnel in working conditions and maintenance operations as described in this manual according to the GENERAL SAFETY WARNINGS This manual contains instructions for professional qualified operators 4 GENERAL SAFETY WARNINGS Use only chemical reagents and accessories specified and supplied by HUMAN and or mentioned in this manual Place the product so that it has proper ventilation The instrument should be installed on a stationary flat working surface free from vibrations Do not operate in area with excessive dust Work at room temperature and humidity according to the specifications listed in this manual Do not operate this instrument with covers and panels removed Only use the power cord specified for this product with th
8. e grounding conductor of the power cord connected to earth ground Use only the fuse type and rating specified by the manufacturer for this instrument use of fuses with improper ratings may pose electrical and fire hazards To avoid fire or shock hazard observe all ratings and markings on the instrument Do not power the instrument in potentially explosive environment or at risk of fire Prior to cleaning and or maintaining the instrument switch off the instrument and remove the power cord For cleaning use only materials specified in this manual otherwise parts may become damaged It is recommended always to wear protective apparel and eye protection while using this instrument Respective warning symbols if appearing in this manual should be carefully considered Diagnostics Worldwide 5 DISPOSAL MANAGEMENT CONCEPT The currently valid local regulations governing disposal must be observed It is in the responsibility of the user to arrange proper disposal of the individual components All parts which may comprise potentially infectious materials have to be disinfected by suitable validated procedures autoclaving chemical treatment prior to disposal Applicable local regulations for disposal have to be carefully observed The instruments and electronic accessories without batteries power packs etc must be disposed off according to the regulations for the disposal of electronic components Batteries power packs and simil
9. e objectives are black anodised Ocular and objectives have an anti fungal coating Vor Figure 6 es Wy em HEI 1 2 28 11 2006 3 56 pm igh b 1 6 10 4 6 Coarse and Fine Focussing Knobs The HumaScope Plus features coaxial coarse and fine focussing knobs Figure 7 on the left and right sides of the instrument The knobs are located below the stage and are easy to operate The coarse and fine focussing range is 30mm The fine focussing knobs are the smaller knobs centred in the coarse focussing knobs The lever on the left inside the left side coarse focussing knob is the coarse focussing stopper When the specimen is in clear focus the stopper may be used to lock in the focus This also allows the stage to be lowered and raised again to exactly the same position When the position is no longer needed simply loosen the stopper Figure 7 4 7 Condenser The Abbe condenser is a specially designed lens mounted under the stage It can be raised or lowered It has an iris type aperture to control the diameter of the beam of light entering the lens system By changing the size of the iris and moving the lens toward or away from the stage the diameter and focal point of the cone of light that goes through the specimen can be controlled Abbe condensers become most useful at magnifications above 400x The condenser lens system should have a numerical aperture equal to or greater than the Numerical Aperture of the objecti
10. ears dark against a bright white background or field 8 10 8 3 Phase Contrast Phase contrast is a technique for revealing the structural features of microscopic transparent objects that cannot otherwise be accomplished with brightfield microscopy Phase contrast achieves the same effect as staining a specimen which can kill a live specimen In phase contrast microscopy the staining is in the optics In a phase contrast microscope the annular rings in the objective lens and the condenser split the light The light that passes through the central part of the light path is recombined with the light that travels around the periphery of the specimen The interference produced by these two paths produces images in which the dense structures appear darker than the background Phase Contrast Light Pathways E 3 5 EE Diagnostics Worldwide 9 10 HumaScope Plus User Manual 8 4 Fluorescence Microscopy The fluorescence of a substance is seen when a molecule is exposed to a specific wavelength of light excitation wavelength or spectrum and the light it emits the emission wavelength or spectrum is of a higher wavelength To view this fluorescence in the microscope several light filtering components are needed Specific filters are needed to isolate the excitation and emission wavelengths of a fluorochrome A bright light source with proper wavelengths for excitation is also needed For normal fluorescence applicati
11. en it is centred open the field diaphragm until its edge is outside the field Your specimen should now be properly illuminated and you should have a good image Figure 4 Diagnostics Worldwide 5 10 HumaScope Plus User Manual 4 4 Mechanical Stage Slide Holder The HumaScope Plus features a double layer two slide stage mechanism See figure 5 This mechanism allows the operator to move the specimen slide along the X or Y axis by turning the corresponding knob This is very useful at higher magnifications as it can be difficult to move the slide in small enough increments by hand Also moving the slide by hand can be difficult since it must be moved in the opposite direction of the observed movement The mechanical stage also has a graduated scale so you can see how far the slide has been moved or keep track of the position of various objects on the slide Figure 5 4 5 Ocular Objective and Nosepiece The microscope imaging system is composed of two oculars and the inward 5 hole objective revolver see figure 6 The objectives are screwed into the threaded holes in the nosepiece which can be rotated to quickly achieve the desired magnification The advanced high precision nosepiece design ensures that the field of view is always centred in the observation area for precise focussing parfocal The ocular tube is inclined 450 for convenient fatigue free observation In order to avoid pertubation caused by reflection th
12. ns by wiping it carefully with a soft clean cloth moistened with alcohol Take care however that alcohol does not seep it into the objectives to avoid dissolving the glue Dust on the lens can be removed using a clean lens brush 3 The coarse and fine focussing assembly and nosepiece are precisely adjusted at the factory and should not be dismantled 4 When not in use keep the objectives in the objective box and cover the ocular tube with the ocular tube cover TE Diagnostics Worldwide 7 10 HumaScope Plus User Manual 7 DISPOSAL MANAGEMENT CONCEPT The currently valid local regulations governing disposal must be observed It is in the responsibility of the user to arrange for proper disposal of the individual components All parts which have come into contact with potentially infectious materials must be disinfected using a valid procedure autoclaving chemical treatment prior to disposal The instrument and electronic accessories without batteries power packs etc must be disposed of in accordance with regulations for the disposal of electronic components Batteries power packs etc must be separated from electric electronic parts and disposed of in accordance with applicable local regulations 8 MICROSCOPE GLOSSARY 8 1 Darkfield Microscopy Darkfield microscopy is a method of acquiring a slide image It is specific to the way the light is projected onto a slide in the microscope equipped with a darkfield
13. ons this is a mercury vapour arc burner E sf E 8 5 Lenses objective lens gathers light from the specimen ocular transmits and magnifies the image from the objective lens to your eye nosepiece rotating mount that holds 4 interchangeable objective lenses tube holds the ocular at the proper distance from the objective lens and blocks out stray light 8 6 Focus to position the objective lens at the proper distance from the specimen coarse focus knob used to bring the object into the focal plane of the objective lens fine focus knob used to make fine adjustments to focus the image 10 10 HUMAN Gesellschaft fur Biochemica und Diagnostica mbH Max Planck Ring 21 65205 Wiesbaden Germany Tel 49 61 22 99 88 0 Fax 49 61 22 99 88 100 e Mail human human de www human de 01 2007 01
14. tness to observe the specimen and make full use of the resolution of the objectives this instrument uses a 6V 20W built in variable brightness lamp For optimal results the condenser must be adjusted properly The condenser is composed of two parts The iris diaphragm is mounted under the stage the illumination source is in the base Adjust the condenser iris mounted under the stage to match the aperture of the objective The stage can be raised and lowered using the focussing knobs The light axis of the condenser must coincide with the light axis of the illumination source To correct the alignment adjust the 3 screws in the part of the condenser mounted on the stage The HumaScope Plus is supplied with an advanced Iris diaphragm It contains an adjustment gear which can be used to adapt the iris setup to the selected objective Figure 3 4 10 4 3 1 Koehler Illumination The HumaScope Plus is equipped with Koehler illumination See figure 4 In order to get the best image possible from the optical setup of the microscope for brightfield phase contrast and differential interference contrast it is crucial that the light path be set up properly HOW TO SET UP HumaScope Plus PROPERLY FOR KOEHLER ILLUMINATION If the light path is set up properly the user has the advantage of an evenly illuminated field a bright image without glare and minimum heating of the specimen The following instructions apply to a microscope equippe
15. ve lens being used 4 8 Immersion Oil Immersion oil is a special oil used in microscopy only with the 100X objective lens usually at 1000X total power A drop is placed on the cover slip and the objective is lowered until it just touches the drop Once brought into focus the oil acts as a bridge between the glass slide and the glass in the lens This concentrates the light path and increases the resolution of the image 5 OPERATION 1 Screw the objectives into the threaded holes in the nosepiece in order of their magnification Insert the oculars into the inclined ocular tube 2 Place a specimen slide on the stage Adjust the stage so that the specimen is centred over the hole in the stage 3 Using first the 10X objective turn the coarse focussing knob to move the objective close to the specimen Now observe the image through ocular and turn the coarse focussing knob to lower the objective until the image comes into approximate focus Next turn the fine focussing knob until the image is sharp Thanks to the instrument s precise and reliable nosepiece a clear focus is maintained even when switching objectives 6 CARE AND MAINTENANCE As with any other optical instrument the microscope should be kept in a cool dry dust and acid free location The instrument should be covered with a dust cover after use 2 Never disassemble the lenses these are precisely adjusted and glued in place at the factory Stains can be removed from the le
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