ey) Polarizing Microscope OPTIPHOT2-POL NA hyBavoid Strong Shocks! Handle the microscope gently, taking ‘care to avoid strong shocks. When Carrying the Microscope ‘When carrying the microscope. support the bottom of the micrascope base. The instrument weighs about 12. kg. Do not hold the overhanging portion of the base. EPlace of Use ‘Avoid the use of the microscope in a dusty place, or where it may be subject to tions, oF exposed to high tempera: tures, moisture, oF direct sunlight, Power Source Voltage and Fuse Check the power source voltage and fuse following the procedure on p.5: Light Source ‘The halogen lamp to be used is 12V- OW. Do notuse bulbs other than the one specifiedin p. 46 (Electrical Specifications), Ifa bulb of mare than the suggested wattage is used, the light adjusting circuit may be damaged [Turning on the Lamp TTake care nat to touch the lamp housing when itis lit, and don’t bring inflammable substances such as gasoline, thinner and alcohol near it, as some parts of the lamp housing may become very hot while the lamp is on __ CAUTIONS _ Thank you very much for purchasing a Nikon Microscope. ‘This microscope is a high precis instrumert with a very delicate structure and varied functions. Please thoroughly read this manual first to use the microscope correctly ion. WChanging the Bulb and Fuse Betore replacing the bulb or fuse, tun OFF the power switch and disconnect the power source plug. Replace the halogen famp (12V-100W), making sure that 1s ool enough. Do not touch the glass pert with your bare hands. Dirt on the Lens Do not leave dust, dirt, or finger marks fn the lens or bulb surtaces. ‘Thy will prevent you from clearly observing the specimen IStrain-free Glasses The sttain-free glasses ere used for the ‘optical elements of this microscope. Handle the objectives and condenser with special care. {Focus Knobs Nover turn the right-or left-hand focus knobs while holding the other -(left- or right-) hand focus knob. It may cause problems, Do not forcibly, turn the focus knobs past the limit because it may cause probiems.CONTENTS: 1 NOMENCLATURE 1 U.ASSEMBLY z 4 Il. MICROSCOPY 13 V.OPERATIONS IN DETAIL 19 1.Oriemation of Dia-polarizer anc Analyzer 19 2. Focusing and Centering the Bertrand lens 20 3. Swing-Out the Top Lens of the Condenser 21 4.1/4 & Tint Plate 24 5 Centering the Objectives “~ 22 6 Stage Rotation, 45° Chp Stop" 23 7 Focusing sven BB 8.Centering the Condenser 25 9. Use of Condenser Aperture Diaphragm 26 10..Use of Field Diaphragm ss 27 11 Optical Path Cnange-Over sv 27 ~~ CARE AND MAINTENANCE [WCleaning the Lenses ‘To clean the lens surfaces, remove dust using a soft brush or gauze. Only when removing linger marks or grease. use a soft cotton cath, lens tissue. or gauze lightly moistened with pure alcohol {iethyt alcoho! or ethyl alcohol. For Cleaning the objectives of immersion oil ‘use only aylene. Do not use xylene for Cleaning the surlace of the entrance lens ‘of the eyepiece tube or the prism surtace of the Ultra-Wide Eyepiece Tube “UW" Observe sufficient caution in handling alcohol and xylene (they are inflammable). and the ON-OFF of the power source switch Cleaning the Painted Surfaces ‘Avoid the use of any organic solvent (for example, thinner, ether, alcohol for Cleaning the painted surfaces and plastic parts of the instrument. We recommend you use the sitcon cloth EiNover Attempt to Dismantle! ‘Never attempt to dismantle the instrument because you may impatr the 12.Intexpupiliary Distance Adjustment z 28 1. Diopter Adjustment: 28 14.011 Immersion Manipulation 29 15. Brightness and Contrast Adjustment 30 16. PHOTO Voltage Set and Adjustment 32 17 Lowering the Substage we 32 18. Objectives and Eyepieces 33 V.OPTIONAL ACCESSORIES ~~~ 34 1 Sénaemont Compensator 34 2.Quarte Wedge 35 3.Pin Hole Eyopiece 36 4, Aitachable Mechanical Stage" 36 5 Universal Stage 37 6 Universal Epiiluminator = 3B. VI.TROUBLESHOOTING 40 ELECTRICAL SPECIFICATIONS ~- 46 When Not in Use When not in use, cover the instrument with the accessory vinyl cover, and store it in a place free from moisture and fungus. It especialy recommended that the objectives and eyepieces be kept in an airtight container containing desiccant EpPeriodic Checking ‘To maintain the best performance of the instrument, we tecommend thet the instrument be periodically checked. (For details ofthis check, contact your ‘surhorized Nikon distibutor. +k Please note as per your Nikon warranty, “Any defects or damage directiy or indirectly caused by the use of ‘unauthorized replacement parts and/or performed by unauthorized pesonnel vl void the warranty1 .NOMENCLATURE Nosepiece centering Centering revolving nosepiat CF Achromat P objective (Strain treo) Specimen clip Vernier Circular graduated stage Condenser centering Condenser aperture diaphragm control ring Field diaphragm control ring Power switch “” duster Lamp housing ‘cover clamp) {indicates also the power som rest ‘switch and PHOTO voltage) L Ames Fig.1-1ns focus fing Analyzer / aterpupillary Analyze Eyepiece Bertrand lens ¢ seraw Diogter ring Analyze 1/42 & tnt plate Coarse-focus stopper clamp ng a Coarse-tocus kno! Vernier Stage rotation clamp screw Fine-focus knob Condenser top lens sming-out knob ‘Achromat swing-out condenser (Strain-free) Power cord condenser focus inoe_f // ip -s condenser camp seew /// Transparent cover ND fiter knob // PHOTO voltage switch Fig. 1-2I1.ASSEMBLY To assemble the microscope, follow the procedures given from Oto ®. For details, read p.5 to p.12. Fig.2 Tools: hexagonal wrench (accessory), plus screwdriver, minus screwdriver, coin, etc.D Check the Power Source Voltage and Fuse @ Disconnect the power cord from the AC mout connector (Assure that itis disconnected.) © Make sure that the line voltage indication plate at the rear of the base indicates the correct line voltage to be used (100-120 or 220-240V) (Fig.2-1) if indicates incorrect voltage, remove the line voltage indic plate by unscrewing the two screws using a plus screwdrwer ‘® Switch over the input voltage change-over switch and reattach the line voltage indication plate so that the voltage ts indicated © Remove the fuse holder at the bottom of the base using a plus screwdriver (Fig. 2-2}. Check the fuse inside by the indication on the metal part of the fuse. —"125V 3.15A” for 100-120V or "250V T2A" for 220-240V— If the incorrect fuse is stored in the fuse holder, please contact your authorized Nikon distributor ‘@ Replace the {use and tightly screw the fuse holder into the original position using a plus screwdrver ® Leveling Foot Screw ‘For stable installation of the microscope, manipulate the adjustment screw at the rear-left of the base (Fig.2-3).® Lamp and Lamp Housing Keep the cover on or use gloves when mounting the lamp bulb so as not to touch the surface directly with your fingers. If the bulb surface is contaminated with finger marks or dirt, wipe them off with lens tissue. Be sure to remove the cover after mounting the lamp. ‘Release the housing cover clamp screw using a coin {f] to remove the cover [2] While pulling down the lamp clamp lever @, firmly insert the lamp Into the sacket pin holes until it reaches the limit @] (Fig.2-4) Caution Be careful not to tilt the lamp when mounting. If tilted, try again Return the lamp clamp lever to the original position ‘®Close the cover and fasten it with the lamp housing cover clamp screw. @ Insert the lamp housing into the lamp housing mount of the microscope base fitting the groove of the lamp housing to the turning limit pin of the lamp housing mount (8) and fasten it firmly | with the lamp housing clamp screw] (Fig. 2-5) a) 4 Fig. 2-5 tion mounting | ‘® Turn off the power switch. (Assure that itis off.) De not touch the bulb immediately after turning it off because it is very hot. Wait Until it cools enough for replacing ‘@To remove the lamp housing, remove the filter holder first, release the lamp housing clamp screw and pull out the lamp housing from the base.EE @ Circular Graduated Stage #Fit the dovetail groove of the substage with the dovetail of the microscope stand, and slide the substage on, Line up the white dot marking (@} on the substage side with the white triangle marking ( on the microscope stand side In this position tighten the substage clamp screw with a minus © screwdriver (Fig. 2-6) ® Specimen Clip Insert the two specimen clips into the holes on the (Fig.2-7) © Revolving Nosepiece Lower the substage by turning the coarse focus knob Loosen the revolving nosepiece clamp screw using the hexagonal wrench. (See ©.) @ install the revolving nosepiece by fitting the groove of the revolving nosepiece with the pin of the circular dovetail, and fasten it with the revolving nosepiece clamp screw [2] (Fig. 2-8) Cautions when dismounting @ Remove the ND filter cassette if attached, and lower the stage by turning the coarse focus knob. Remove the objective if attached #When loosening the revolving nosepiece clamp screw support the revolving nosepiece 80 it does. not drop lage surface@ Objectives ‘# Screw the objectives into the holes of the revolving no positions such that, when viewed from above, their r Power increases as the nosepiece is revolved clockwise (Fig 2-9) [Cautions when dismounting | #Remave the NO filter cassette if attached, and lower the stage by turning the coarse focus knob #Remove any specimen from the stage Hold the objective so as not to drop it when unscrewing ® Condenser ‘fiaise the substage by turning the coarse tocus knob . Completely lower the condenser carrier by turning the condenser focus knob ‘Insert the condenser into the condenser carrier [1] with the aperture number plate facing toward the user, Fasten it with the condenser clamp screw @] (Fig.2-10) ‘Completely raise the condenser carrier by turning the condenser focus knob [Cautions when Dismounting ‘*Flemove the ND filter cassette if attached Completely lower the condenser carrier by turning the condenser focus knob. Remove the condenser by releasing the condenser clamp screwSe @ Intermediate Tube ‘© Mount the intermediate tube on the microscope arm by fitting the groove of the circular dovetail of the intermediate tube bottom with the tip of the tube clamp screw. Tighten the clamp screw using the hexagonal wrench. (See @ and Fig. 2-11} (@ 1/42 and Tint-Plate ‘@ Remove the screw from the side of the 1/44 plate. Insert the plate nto the compensator slot of the intermediate tube facing 1! Positioning notches toward you. Reattach the screw. (Fig. 2-12} @ Trinocular Eyepiece Tube ‘Mount the eyepiece tube onto the intermediate tube by fitting the groove of the circular dovetail of the eyepiece tube with the tip of the clamp screw. Tighten the clamp screw with the hexagonal wrench (See @ and Fig. 2-13) Cautions when clamping ‘Tighten the eyepiece tube clamp screw holding the stem of the hexagonal wrench. If overtightened holding the plastic part, optical-path change-over of the eyepiece tube may be malfunctionedSLL (2 Eyepiece ‘® Use the same magnification eyepieces for both the right and left eyes @ Insert the eyepi ces into the sleeves of the observation tube by engaging the three grooves of the eyepiece with the three protrusions of the sleeve (Fig. 2-14) When using eyeguard rubbers, put them onto the eyepieces (Fig. 2-15) J Fig.2-14 (3 Filter (Filter Holder) When mounting/dismounting the filter to/from the filter holder, use gloves or gauze so as not to touch the filter with your bare hands. Mount the filter 10 be used as shown in Fig.2-16-], NCB11, GIF. and ND filters should be mounted to match their respective indications Insert the filter holder into the filter holder mount (Fig.2-17). Leave the three filter holders inserted to protect the holder mount from dust even when the filters are not mounted When removing the filter, push the filter back in the same direction it was attached as shown in Fig. 2-16- 101 @ND Filter Cassette Push down the ND filter cassette so the two protrusions of the cassette bottom fit with the two mounting grooves on tt field lens part (Fig. 2-18) ‘When frequently lowering the stage, mount the ND fier cassette in the direction shown in Fig.2-19 ‘®When dismounting, push the cassette either to the lett or right and raise the opposite side rim of the @ Power Cord Insert the socket of the power cord to the AC input connector and plug it into an AC line receptacle (Fig. 2-20) When the receptacle is not a double-pole ground type, use a plug converting adapter (Fig. 2-21) Ground Sexes Fig.2-21is stored in the ack of the microscope 1213 I. MICROSCOPY 1) Turn ON the power switch to light the lamp (1). Set the lamp brightness indicator to "9" using the brightness adjuster (2) or by turning the PHOTO voltage switch to ON (Fig.3-1) 2) Pull out the analyzer knob of the intermediate tube [1] to remove the analyzer from the optical path, Turn the Bertrand lens turret to the “0” position 2) to remove the Bertrand lens out of the optical path (Fig. 3-2) 3) Slide the ND filter knobs of the ND filter cassette to the limit to place all the ND filters in the optical path (Fig. 3-3) 4) Put the NCB11 filter (or necessary filter) into the optical path (Fig. 3-4)5} Revolve the revolving nosep ece to swing in the 10x objective to the optical path. Assure that the nosepiece properly settles in position (Fig.3-5) 6) Place the specimen (glass slide) on the stage with its caver giass facing up and fasten it with the two specimen clips (Fig.3-6) 7) Raise the condenser uppermost using the condenser focus knob (Fig 3-7) - 8) Fully open the fiald and the aperture diaphragms (Fig 3-8) 1415 | Object portion ‘enthe ‘optical path. 9) Bring the object portion of specimen into the optical path (Fig.3-9). {Object portion is immediately above the condenser lens.) 10) Loosen the coarse-focus stopper clamp ring until it reaches the limit (Fig.3-10) (See p.24, Operations in Detail-7-3).) 11) Push in the optical path change-over knob of the eyepiece tube to enter 100% Of the light into the ‘observation tube (Fig. 3-11) (See p.27, Operations in Detail-11.) 12) Focus on the specimen locking in the eyepiece by manipulate the coarse/fine focus knob (Fig. 3-12)13) Adjust the diopter (Fig.3-13) (See p.28, Operations ir Detail-13.) 14) Adjust the interpupillary distance (Fig. 3-14), (See p.28, Operations in Detail 12.) 15) Center the objective (Fig.3-15). (See p.22 Operations in Detail-5.) 1617 16) Focus and center the condenser (Fig.3-16). (See p.25, Operations in Detail-8.) 17) Push in the analyzer knob to Eyeniece put the analyzer into the @ vowed stop aptical path (Fig. 3-17) 18} Revolve the revolving nosepiece to the objective to be used and focus on the specimen by manipulating the fine or coarse focus knob (See Note: 2.) Wien using an oil immersion objective, apply oil to the space between the top of the objective and the specimen Take care not to produce bubbles in the oil. See p.29, Operations in Detail-14.) 19) Adjust the brightness by sliding the ND filter knob of the ND filter cassette. (See p.30, Operations in Datail-15.) When adjusting the brightness of the lamp, turn OFF the PHOTO voltage switch and manipulate the brightness adjuster 20) Adjust the viewfield and the aperture diaphragms by manipulating their respective control rings. (See pp.26 and 27, Operations in Detail-9 and -10.)Table 1 Orthoscopic microscopy Conoscopic microscopy Top tens of Bertrand lens Tox or | higher IN toner’ our our IN | iN Aperture diaphragm Field diaphragm 70% 80% of she Cicumscribed the circumference of the Fully opened Fully opened eyepiece fad ot" ‘conoscopic field of view {ar fully opened) Circumseribed the circumference of the orthoscopic field of view Manipulate the viewfield is dark items # Insertion/removal of the ND filter @ Turning the revolving nosepiece (click-stop position) ‘Position of the condenser @Viewfield and aperture diaphragms fully open #Change-over of the optical path of the eyepiece tube @ Mounting the lamp © Mounting the revolving nosepiece ‘Mounting the condenser © Mounting the filter holder Check the following items when the focused Placing the specimen © Releasing the coarse-focus stopper clamp ring Position of the substage ‘© Thickness of the cover glass (standard =0. 17mm) condenser centering screws if part of the If this doesn’t help, check the following Il, MICROSCOPY-3) Il MICROSCOPY-5) m0 iO MICROSCOPY-7) MICROSCOPY-8) MICROSCOPY. 11) ASSEMBLY-@ Mounting the lamp and the lamp housing ASSEMBLY-© Mounting the revelving nosepiece ASSEMBLY-® Mounting the condenser ASSEMBLY-@ Mounting the filter 0 uw specimen cannot be ii ii MICROSCOPY-6} MICROSCOPY- 10) 11.ASSEMBLY-© Mounting the circular graduated stage 1819 IV. OPERATIONS IN DETAIL soseon a as 1. Orientation of Dia-polarizer and Analyzer (Dia-polarizer and Analyzer) When the dia-polarizer and analyzer are set at “O” on the protractor scale, position of the polanzation plane coincides with the orientation plate on the microscope base, offering the crossed Nicols (dark eross) Position. The plate shows the indication “P* of the X-direction is for dia-polarizer and the “A” of the Y-direction is for analyzer. (Fig. 4) Note | Some reference books for polarizing microscope available in the market may explain that the X direction is for analyzer and the Y-direction, for polarizer Fig © Dia-polarizer attached on the bottom of the substage is rotatable 36 and detachable by pulling down. (Fig.6} When attaching, align the positioning pin on the dia-polarizer with the groove at “O™ position of the bottom of the substage The analyzer can be rotated by loosening the analyzer clamp screw [1] and rotating the rotation ring @J. The rotation angle can be read in the range between 0° to 180" with the unit of 0.1" by the vernier. The analyzer can be removed from the optical path by pulling the analyzer knob Gl. (Fig.6) Note ] As the depolarizer is —- builtin the intermediate tube, you don’t need to be concerned about the relation between the orientation of the polarizing plate and photomicrographic deviceIN DETAIL 2. Focusing and Centering the Bertrand Lens (Bertrand lens of the intermediate tube) When objectives are replaced, turn the Bertrand lens focus ring under the the Bertrand fens turret and focus the Bertrand lens Centering is carried out using the condenser aperture diaphragm image and the two centering screws. (Fig.7) Centering procedure for the Bertrand lens is the same as that for the condenser except that the condenser aperture diaphragm image is used in place of the field diaphragm image in condenser centering (Refor to Item 8.) [Note] ¢Another Bertrand lens is built in the trinocular eyepiece tube. Turn the Bertrand lens ring to the position of "8". and the Bertrand lens will be brought into the optical path and a conoscope image may be observed, (Fig.8) 20A a 3. Swing Out the Top Lens of the Condenser (Swing-out condenser) ‘®The top lens of the condenser can be removed from the optical path by the swing-out knob. (Fig. 9} Bring the top lens into the optical path during normal orthoscopic microscopy or conoscopic microscopy. Swing it out for orthoscopic microscopy with 4.x or lower magnification objective When measuring the retardation or observing the interference color. ‘swing out the top lens (or stop down the condenser aperture diaphragm) so as to make the illumination light flux as parallel as possible to the optical axis 4. 1/4, & Tint Plate @The 1/42 & tint plate has an empty hole at the center, By pushing it into the compensator slot, the sensitive tint plate (530nm) is brought into the optical path and by pulling it out the 1/4 plate is brought into the optical path. It is used for recognition of very weak birefringence and the determination of X" and Z’ of the specimen 21PAS 5. Centering the Objectives (Centering revolving nosepiece, Circular graduated stage) Before centering the objectives, implement the microscopy procedures from 1} to 14) so that a specimen is focused using the 10x objective Make coincidence with the rotation center of the stage and the cross lines center of the eyepiece by the following procedure @Bring an appropriate target on the specimen to the center of the crass lines in the eyepiece insert the centering tools in the centering screws on the nosepie: (Fig. 10) @Rotate the stage about 180°, and the target is displaced from the center of the cross lines. Move the objective using the centering tools. 0 that the center of the cross lines moves one half the displacement of the target. (Fig. 11) Repeat the above procedure two or three times. Carty out centering for each objective Saplacerant| woe Tata A 2223 CE ey Ss 6. Stage Rotation, 45° Clip Stop (Circular graduated stage) ©The stage can be rotated by loosening the stage rotation clamp screw. The rotation angle of the stage is readable with the accuracy of 0.1" via paired vernier scales, When the reading with one of the vernier scales is interrupted by the attachable mechanical stage (option), read the other vernier and add +90" to the reading @The 45° click-stop device of the stage is effectuated at every 45° starting from a position where the click stop lever has been pulled toward you. This offers convenience in switching the observation over from a crossed Nicols position to a diagonal position. (Fig. 12) top release, turn the lever toward the microscope stand 7. Focusing 1) Coarse and Fine focus (Coarse and Fine focus knobs) ‘© Focusing is carried out by the coarse and fine focus knobs at the left and right of the microscope stand. ©The direction of vertical movement of the stage corresponding to the turning direction of the focus knob is shown in Fig. 13, ‘© One rotation of the fine focus knob moves the stage 0. Imm and the graduation on the fine focus knob is 1 micron. One rotation of the coarse focus knob moves the stage 12mm. The range of coarse and fine motion is 2mm up and 28mm down from the standard position Never turn the right or left knob while holding the other. It may cause problems. Do not tum the coarse and fine focus knobs further than the lio 2) Coarse-focus torque adjustment (Coarse-focus torque adjustment ring) The rotation of the coarse-focus knob can be tightened when the torque adjustment ring is turned counter-clockwise 3) Coarse-focus stop (Coarse-focus stopper clamp ring) Turn the coarse-focus stopper clamp ring and clamp the coars movement at the position where the specimen is focused. It is clamped at the position where the black triangle mark (d\} on the side of the clamp ring reaches the top. (Fig. 14) @When the coarse-focus stop is carried out, the stage cannot be raised from that position using the coarse focus knob. However, the fire focus knob can move the stage regardless of the limit © Lower the stage by using only the coarse focus knob and replace the specimens. © Raise the stage slowly unti it reaches the limit using only the coarse focus knob. The specimen should be almost focused at the position where the stage reaches the limit. Then focus accurately by turning the fine focus knob When the coarse-focus stop is not used, loosen the coarse-focus ‘stopper clamp ring until it reaches the limit. (See Fig. 3-10) focus 2425 8. Centering the Condenser (Condenser focus knob, Condenser centering screw, Field diaphragm control ring) (Before focusing and centering the condenser. implement Microscopy procedure from 1) to 16} and focus on the specimen with 10% objective.) Close the field diaphragm to its smallest size by manipulating the field diaphragm control ring. Rotate the condenser focus knob to move the condenser vertically so that a sharp image of the field diaphragm is formed on the specimen surtace (Fig. 15 a Eyepiece viewed sop Eyopioce viwsold stop Fig.15 @If the image decenters from the viewlield of the eyepiece, bring it roughly to the center of the viewtield by moans of the condenser centering screws (Figs. 15-2) and 16) @change to the 40 objective. Focus on the specimen by turning the fine focus knob and form an image of the “ield diaphragm on the specimen surface by manipulating the condenser focus knob @when the image decenters from the viewliald of the eyepiece, bring it to the center of the viewfield by means of the condenser centering screws. At this time, stopping down the field diaphragm image to be slightly smaller than the viewfield of the eyepiece may be convenient for centering (Fig. 15-3)POSEY 9. Use of Condenser Aperture Diaphragm (Condenser aperture diaphragm control ring) 1) For orthoscopic microscopy The condenser aperture diaphragm is provided for adjusting the numerical aperture (N.A.) of the illuminating system of the microscope It is important because it determines the resolution of the image, contrast, depth of focus, and brightness Stopping down the aperture diaphragm will lower the resolution and brightness but increase the contrast and depth of focus. In general when itis stopped down to 70% ~80% of the numerical aperture of the objective, a good image of appropriate contrast will be obtained (Fig. 17) Ullal — Exit pupil of objective >, 1020 | “05 Jp -reorse tephra Size of the condensar aperture diaphragm Fig.17 To adjust the size of the condenser aperture diaphragm, manipulate the diaphragm control ring referring to the condenser's N.A. scale, or observing the diaphragm image visible on the exit pupil inside the objective. The exit pupil can be seen by removing the eyepiece from the eyepiece tube or putting the Bertrand lens into the optical path. Stopping down the aperture diaphragm too far will lower the resolving power Therefore, it is recommended not to stop down the aperture to a size smaller than 60% of the NA. of the objective in use, except when observing almost transparent specimens. When swinging out the top lens of the condenser (for microscopy using 4X or lower objective), fully open the condenser aperture diaphragm 2) For conoscopic microscopy In conoscopic microscopy, the condenser aperture diaphragm works as a field diaphragm on the conoscopic image surface. Stop down the diaphragm to such an extent that it circumscribes the circumference of the field of view of the conoscopic image (exit pupil of objective) to shut out the stray light 2610. Use of Field Diaphragm (Field diaphragm control ring) The field diaphragm is used for determining the illuminated area on the specimen. The size of the diaphragm is adjusted by manipulating the field diaphragm control ring. In general stop down the diaphragm to such an extent that the circumference of the illuminated area circ or inscribes that of the eyepiece field of view. if a wider area 1! required is illuminated, extraneous light will entor the field of view. causing flare in the image and lowering the cortrast. Therelore especially in photomicrography. the proper adjustment of the field diaphragm is very important. Generally, good results will be achieved when the diaphragm is stopped down to such én extent that the diameter of the illuminated area is slightly larger than the diagonal of the film format ‘@ Thickness of the glass slide must be 1.7mm or less otherwise, the field diaphragm image may not be focused on the specimen surface. ®This diaphragm does not work as the field diaphragm when the condenser top lens is swung o.t of the optical path. In this case, fully open the diaphragm as the numerical aperture of the illuminator will be cut off when this, diaphragm is excessively stopped down 11. Optical Path Change-Over (Optical path change-over knob of the trinocular eyepiece tube) ‘As shown in Fig, 18, when the change-over knob is pushed until it reaches the limit [1], 100% of the light enters the observation tube. When the change-over knab reaches the intermediate click (2). the proportion of light entering the observation tube and photo tube will be 14:86. When the change-over knob is pulled to the limit 3), 100% of the light enters the photo tube. Fig. 18PASS STST.T 12. Interpupillary Distance Adjustment (Observation tubes) (Before adjusting the interpupillary distance, implement MICROSCOPY procedure 1}t0 13) and facus on the specimen with the 10x objective. ‘Adjust the interpupillary distance, so that both the right and left viewlields become one (Fig This adjustment will enable the user to observe the specimen with both eyes Fig.19 13. Diopter Adjustment (Eyepiece) (Before adjusting the diopter, implement MICROSCOPY procedures 1) to 12) and focus on the specimen with the 10 x objective.) Make diopter adjustments for both the right and left eyepieces ©Turn the diopter compensation rings on each eyepiece until the end surface of the ring coincides with the engraved line. {This is the position of O dioptic compensation.) (Fig. 20-1) @Swinging the 40x objective by turning the revolving nosepiece and bring the specimen image into focus by turning the fine focus knob (or the coarse focus knob) Ss nd surface cw tos & Engraved kine Fig.20-129 @Swing the 4x or 10 objective into position. Without manipulating the fine and coarse focus knobs, turn the diopter rings on the eyepieces so that the specimen images in the right and lett eyepieces are focused individually (Fig. 20-2) ‘# Repeat the above procedure two times. to adjust the diopter perfectly # The above adjustment, compensating diopter difference between the User's right and left eyes, will Keep the tube length of the microscope correct. This enables the user to take full advantage of the high-quality objectives, including their parfocality 14. Oil Immersion Manipulation (Oil immersion objective) The CF achromat P objective 10x is an oil immersion type and is to be immersed in oil between the specimen and the front lens of the objective. Use only the included oil Apply the minimum amount necessary (the quantity that fils the gap between the front of the objective and the spesimen) to avoid a fiow of excessive oil that will adhere to the stage To see if air bubbles are present in the immersion oil, which deteriorate the image quality, pull out the eyepiece from the eyepiece tube. Fully open the viewlield diaphragm and the aperture diaphragm to examine the objective exit pupil (bright circle) inside the tube: To remove air bubbles, slightly rotate the nosepiece several times, or apply additional oil, or replace the oil. Any remnant of oil left on the oil immersion objective or adhesion of oil to the front of the dry system objective will deteriorate the image quality Clean off the oil after using it and make sure that the oil did not adhere 10 the front of other objectives To clean off the oil, wipe with lens tissue or a soft cloth, moistened with xylene, lightly two or three times over the lens. Itis essential at this time to avoid touching the lens with a part of tissue or cloth already used.PASS 15. Brightness and Contrast Adjustment (ND filter cassette and Filter holder) ND filters supplied with the ND filter cassette are used for brightness adjustment in general microscopy and photomicrography illumination brightness can be reduced to 1/2—1/128 by the combined use of these filters. (Reter to Table 2.) Securely change the filter knob when inserting or removing the filters Less frequently changed filters such as shown in Table 3 are used attached to the filter holder. ND filters can be also attached to the filter holder when it is too bright with only the ND filters of the ND filter cassette used Any filters of 45mm in outer diameter and 3mm or less in thickness can be mounted to both the ND filter cassette and filter holder Filters in ND Fitter Cassette Table 2 N Transmission Light Reduction by Fiter Combination Rate ‘pe. “Does. pes. N02 | Abour6o% | 1/2 A 1/8 nos | Abou 25% | ssi/a 21/32] 1/128 1/64 wp16 | About 6% e116 Filters in Filter Holder Table 3 Name Purpose Use ] NCB1T [Color balancing For ganeral microscopy and color Retardation measurement, |For phase-contrast meroscopy =| or Contrast adjustment monochrome photomicrograph 30"ERATIONS IN DETAIL Removing and mounting the ND filters Use gloves or gauze so as not to touch the filters with your bare hands. Lay soft cloth such as gauze on a desk beforehand. Place the ND filter cassette on it, spread the lever and remova the filter (Fig.22} When mounting, insert the filter obliquely in the opposite side of the lever and spread the lever from the lower side Fig.22 Transparent Cover Place the transparent cover on the ND filter cassette to protect the filters from dust (Fig. 23-1) When the ND filter cassette is removed, place the transparent cover on the field lens (Fig. 23-2) However, remove the cover during observation or photomicrographing with the viewfield diaphragm stopped down 3116. PHOTO Voltage Set and Adjustment (PHOTO voltage switch and PHOTO voltage fine adjustment switch) A constant voltage should be used for color photomicrography because the color temperature of the lamp varies with the voltage being used The color temperature balancing filter (NCB1 1) should be used when daylight-type color film is used. The standard ts to set the voltage of the amp to 9V. The PHOTO voltage switch is provided to set the standard lamp voltage automatically, The lamp voltage is set to 9V, when the switch is turned ON, regardless of the position of the brightness adjuster Fine adjustment of the PHOTO voltage The PHOTO voltage of 9V can be finely adjusted by about ~0.9¥. +0.8V, or +1.6V. When the color film photographed at 9V is reddish, the voltage should be raised. (A higher lamp temperature gives a bluish color.) When bluish, the voltage should be reduced (A lower lamp temperature gives a reddish color.) Turn the rotary switch at the bottom of the base with a minus screwdriver to adjust the voltage (Fig.24) When this adjustment is inadequate. also use color compensating fiers (CC filters) Fig.24 17. Lowering the Substage (Substage) The mounting position of the substage can be lowered by releasing the substage clamp screw with a screwdriver. It is lowered as far as 32mm down from the standard position for observation, permitting the use of the universal stage (optional) and the observation of the thicker specimen (mainly in episcopic polarizing microscopy} 3233 EE ey ER 18. Objectives and Eyepieces In the OPTIPHOT2-POL, the CF P objectives (strain-free) and CF evepieces based on the CF (Chromatic Aberration Free) system are adopted. Always use the CF objective and the CF eyepiece in ‘combination. Since the compensation of chromatic aberration is carried out respectively in the objective and the eyepiece. the direct image produced by the objective can be utilized for observation by a TV camera, otc With the objectives marked "160/0.17", use a covergiass of 0.17mm in thickness. The indication “160/—" on the objective means that no matter whether a covergiass is used of not, no decrease of image definition of of contrast will result ‘When the eyepiece with cross lines and scale is inserted into the eyepiece sleeve with the orientation pin fitted to the right-hand groove of the sleeve, the O-direction of the analyzer and dia-polanizer are aligned with the cross lines direction If the orientation pin is fitted to the upper-right groove of the sleeve, the cross lines will be aligned with the diagonal position of the polarization ‘CF PL Projection lenses are exclusively designed for photomicrography Do not use them for observation. '®For focusing with the binocular observation tube in photomicrography, use the eyepiece with photo mask.V_. OPTIONAL ACCESSORIES 1. Sénarmont Compensator To be inserted into the compensator slot af the intermediate tube in place of the 1/42 and tint plate to measure the retardation with the accuracy of the lambda (a) unit. (Fig. 26) Fig.26 1) Detecting extinction position Rotate the stage with the specimen under the crossed Nicols to find out the direction where the part of the specimen to be measured appears darkest 2) Detecting subtraction position Rotate the stage 45° to bring it from the extinction position to the diagonal position. Insert the 1/4 and tint plate into the optiéal path, and confirm that the interference color of the part of the specimen to be measured changes toward the lower order side. if the color changes toward the higher order side, rotate the stage another 90" 3) Mesurement Place the GIF filter on the filter receptacle, and replace the 1/44 and tint plate with the compensator. Rotate the analyzer so that the part of the specimen to be measured becomes darkest Let the angle of the above analyzer rotation be theta (é] then the retardation R (nm) will be obtained as follows: ay 780 Where a: wave length of the light used for the measurement When the GIF filter is used: A=546nm 34ETO 2. Quartz Wedge To be inserted into the compensator slot of the intermediate tube instead of the 1/44 and tint plate. (Fig.27) The quartz wedge is engraved with scale and used for the rough ‘measurement of retardation in the range of 1A~6A. Fig.27 1) Detecting extinction position Detect the position where the part of the specimen to be measured becomes darkest by rotating the stage under the crossed N 2) Detecting subtraction position Rotate the stage 45° to bring it from the extinction position to the diagonal position. Insert the quartz wedge into the optical path, and confirm that the interference color of the part of the specimen to be measured changes toward the lower order side If the color changes toward the higher order side, rotate the stage another 90° 3) Measurement Slide the quartz wedge along the stot, and the interference color changes Stop sliding the quartz wedge where the dark stripe covers the part of the specimen to be measured. In this position. find the part with no object under the same dark stripe, and compare the interference color of that part with the Interference Color Chart to assume the amount of retardation If the view field around the part to be measured is entirely filled with the object, restrict the illumination to the part to be measured by means of the field diaphragm, remove the specimen from the optical path and then compare the interference color with the Chart 35ONS 3. Pin Hole Eyepiece Replace one of the paired eyepieces with the pin hole eyepiece. Remove the Bertrand lens out of the optical path, and the conoscopic image can be observed overlapping the orthoscopic image through the binocular observation. (Fig.28) In the above simultaneous observation of both images, the conoscopic image may be observed deviated from the orthoscopic view field center The conoscopic image observed through the pin hole eyepiece is represented by the conoscopic light flux that covers the central part of the orthoscopic view field to the extent of about 1/20 4. Attachable Mechanical Stage - Mount the attachable mechanical stage on the circular graduated stage inserting the two positioning pins on the bottom of the attachable stage into the two pin holes on the graduated stage surface. Tighten the clamp screw using a screwdriver or coin. (Fig.29) The attachable mechanical stage with point counters (whose pitch are 0.2mm or 0.3mm) is also available. The point counter can be replaced by releasing the head of the point counter using a coin and removing the milled part of the counter. To release the click-stop of the point counter, release the click spring nut ex e Positioning 3637 Ce 5. Universal Stage The universal stage can be used for the polarizing microscope OPTIPHOT2 POL. When using the universal stage, lower the substage beforehand to face the white dot @ with the mark €T on the microscope stand. (Refer to p.32-17,) For the usage of the stage, refer to the separate instructions on Universal Stage’ sage ZIV eoton | fempnn eV) Nemphes oe6. Universal Epi-illuminator Used for episcopic polarizing microscopy, mounted between the OPTIPHOT2-POL stand and the intermediate tube. 1) Nomenclature and Assembly Referring to Fig. 31, assemble in the order given. CDRemove the eyepiece tube and the intermediate tube from the microscope stand @Mount the universal epi-illuminator on the microscope arm positioning the illuminator nearly parallel to the arm, Clamp the screw @Attach the lamp bulb (12-50W Halogen lamp) and socket to the lamp housing. Release sufficiently the clamp screw on the lamp housing and mount it to the universal epi-lluminator. Retighten the clamp screw. @onnect the lamp cord to the transformer @Remove the accessory ND32 filter slider from the illuminator. Insert the polarizer slider into the illuminator until the slider clicks twice. @Place the filters. @Mount the intermediate tube onto the illuminator, fitting the notch of the circular dovetail with the end of the clamp screw. Fasten the clamp screw @®Referring to p.9, mount the eyepiece tube on the intermediate tube. C= > 2 Lamp housing clamp screw Socket soeve clamp screw ‘Aperture diaphragm sin Field ciaphragm ing Univers Lamp lateral entering serew ranstorner Lamp vertical cantonng fing olazer sider intermediate tube clam serew iaceone a Opticat- path change-over knob Dust-ight sider ~Fig.31 3839 A YEE 2) Preparation (1) Centering the lamp Make certain that the optical-path chang the limit Turn ON the power switch on the transformar and set the voltage to 6V @Fully open the aperture diaphragm. @Place the ND filter on the stage and focus on it using the 10x objective Remove the eyepiece from the sleeve (or put the Bertrand lons into the optical path), and looking into the exit pupil of objective, move the lamp housing back and forth to form a sharp image of the lamp filament on the diffuser of the exit pupil @Manipulate the lamp centering screws to center the filament image on the exit pupil (2) Orientation of polarizer Roughly focus on the ND fier on the stage using the 40x objective @Set the analyzer scale to "0" Bring the Bertrand lens inio the optical path. Looking into the exit pupil of the objective. rotate the polarizer rotation ring to form the dark cross image on the exit pupil. (Fig.32) Note ] Take care not to touch the polarizer rotation ring while observing the specimen, or the orientation of the polarizer will have problems It itis touched by mistake, readjust the orientation 7 ‘over knob is pushed in to LU Dark cross image Fig.32 3) Objectives Use the CF M Plan Achromat P series objectives (Strain-free, 210/45) For microscopy, refer to the descriptions for diascopic polarizing microscopyVI. TROUBLESHOOTING SS SEEING AND OPERATION Failures Causes I ‘Actions (Opucal path in tinocular tube not tully changes Changeover the path securely to Optical path #9 tinooular binocular observation changed: over for binocular tube 27 observation Revolving nosepiece not mounted | eer Mount tiem 67 Revolving nosepiece notin Revolve it to click stop lick-stop position (objective not | position (put objective centered in optical ath) into optical path). (Ip 14) Position correctly so the Condenser is too low wiewield diaphragm image is formed. (p 25) Darkness at the | Condenser not centered Centering periphery Condenser not mounted correctly. | Mount correctly Uneven IND fiter not fully changed-over. | Changeover to the limit brightness of Sot NY Crenenover_| Crengeover to Viewticl. ‘iter holder not mounted correctly. | Mount property in Fiter holder not mounted "| comeet postion. (p.10) No appearance | Fieid disphragm closed toa much. | Open properly (p 271 of viowfeld Improper combination of objective Improper come Use proper combination Dirt or dust on the lens (field lens. condenser. objective, eyepiece. eanin >. eyepiece tube entrance lens) or | ean. oo specimen, Lamp not mounted correctly Mount properly. _(p.6) Fomove out of th Bertrand lens in optical path, cee ie a3) Top fens of the condenser not fully | Swing in or auto the swing outlin iri 20 1/44 and tint plate or compensatar not in correct positian Insert property. (p.21)41 ES Failures Dirt or dust in the viewtield Poor image obtained (Contrast is too strong or too) weak.) {Details are not clear.) Causes, Position af condenser to0 low Aperture diaphragm toa restricted Dir ar dust on the lens (el lens condenser, objective, eyepiece, eyepiece tube entrance len: ecimen I Position of condenser 100 low. Position correctly so the Actions Position correctly s0 the Vviewlield diaphragm image is formed. (p.25) Open properly. _{p.26) Cleaning ow Open properly. (p.26) Viowfield diaphragm image is formed. (p.28) Too thick oF thin coverglass. No coverglass attached NG objective for observing specimen without coverglass used to ‘observe specimen with coverglass Normal objective for observing specimen with coverglass used 10 observe specimen without coverglass: Use specified thickness (0. 17mm) coverglass Use normal abjective for observing specimen with coveralass, Use NCG objective | 'No immersion oil used on the front of immersion system objective Immersion oil used not the ne spectied ‘Air bubbles in immersion oil Use Nikon immersion | oi (o.29)} Remove bubbles. (p.29) Immersion oil soils the top of ary system objective (especially 40%) Cleaning (».29) ‘Compensation ring of objective not adjusted ‘Adjust to match coverglass, Dirt or dust on the lens (field lens: condenser, objective, eyepiece, eyepiece tube entrance lens) or specimen, Cleaning fo.Image too bright, Not bright enough (see also Electrical). Lamp power source voltage too | high. Lamp power source voltage 100 ‘Aperture diaphragm too restricted. Position of condenser 100 low. ‘Optical path change-over of ‘rinocular tube not 100% binocular Fale Causes rion | | image is dim. Place it stably by ety Revohi tach coveety. 9. | click-stop position. position (p14) | Image moves it su - Paces by focuses (p.14) centered er 9 {p,28) Sige ed ‘iach coneaiy. 0.0 NCBI ter wat od Use NCBI her 13) image tinged | use NCB He bd Tam power soueevolage too | Saiamp valage 6 by turning ON the PHOTO voltage switch or by manipulating the brightness adjuster. Use the ND fiters of the low. | cassette, (p.13 & 30} ‘Open properly. _(p-26) Position correctly 80 the viewfield diaphragm image is formed. (p.25) that 100% of light enters binocular part. (p.27) 4243 Failures No focused image obtained with high powers | objectives, | Slide upside-down Covergiass too thick Actions Use covergiass of specified thickness (0.1? ich to stage with covergiass up (when no coverglass, specimen surface should be up) 14 a | High-power objective touches the specimen, when changed-over | from low power Insufficient | parfocality of objective when changed-over. Slide upside-down Coverglass to0 thick, Eyepiece diopter not adjusted Eyepiece diopter not adjusted ‘Attach ta siage with covergiass up (when no coverglass. specimen surface should be up) Use coverglass of specified thickness (0.17mm) Diopter adjustment Diopter adjustment 14) (p.28) (9.28) Movement of image not smooth when moving the | specimen. (When optional | attachable mechanical stage is used.) Attachable stage not tightly fastened | to stage. Fasten it tightly 36) Travel of stage limited to one-half length of slide. (When optionat attachable mechanical stage is used.) Improper attachment of attachable stage Shift the attachment | position (.36)ESS [Failures _ Causes ‘Actions ‘Adjust nterpupsllary Interpupiary astance not adjusted, | Aone merPup No cohesion of — binocular image Diopter adjustment Eyepiece diopter not adjusted ‘p.2m) Magnifications of aight and left oyepieces ater Use same eyepieces ‘Adjust inte Intorpupillary distance not adjuste p i " distance. Diopter adjustment 28) Use the ND fier cassame nacequata ilumination, | and adjust the brightness | Inea Harn with combination of ND fiters (9.301 | Eyepiece diopter not adjuste Experiencing eve | Eyepiece diopt usted fatigue. 4445 ELECTRICAL Failures Brightness of lamp does not ‘change even though brightness adjuster is ‘manipulated Lamp does not light even though switched ON. PHOTO voltage switch is ON No electricity obtained [No bulb attached Bulb blown, Fuse blows Lamp a correctly mounted Jtemrworo ctoge | puron or 3a)| Connect the cord to socket Attach lamp 6) Replacement. (p 5 Flickering and unstable illumination. ment, (p.6) Bulb about to blow. } " Connector not secure. [Fuse holder not firmiy fastened. Connect power source Cord and lamp housing securely. (p.6 and p.11) Fasten securely. (p 5) Bulb insufficiently inserted into the ‘Bulb immediately blown. Bulb used not the one specified Fuse used not the one specified Use 12V-100W halogen | lamp OSRAM HUX 64623 or PHILIPS Insufficient 7724 |itimination, rel [Mumia input voltage change-over switch | Changeover input does not match the power votage of | voltage change-over the room switch (5) Fuse blown, —Vncms > Use the fuse specified 0.51]| | | ELECTRICAL SPECIFICATIONS 100-120V/220-240V, 50/60Hz 12V 100W (use OSRAM HLX 64623 or PHILIPS 7724) ~ 100-120V:125V 3.15A 220-240V:250V T2A soe less than 130W Power source Halogen lamp -~ Fuse Power consumption - REFERENCE This manual instructs only how to manipulate the OPTIPHOT2-POL microscope For the practical explanation on polarizing microscopy. refer to the following special works ‘AN INTRODUCTION TO THE METHODS OF OPTICAL CRYSTALLOGRAPHY” — F. Donald Bloss Holt, Rinehart and Winston “ORE MICROSCOPY” — Eugene n. Cameron John Wiley & Sons, Inc. ‘© “THE POLARIZING MICROSCOPE” ‘A. F. Hallimand Vickers Instruments Nikon reserves the right to make such alterations in design as may be considered necessary in the light of experience. For this reason, particulars and illustration in this, handbook may not conform in every detail to models in current production, 46