AZ-STD Diascopic Stand and AZ-STGD DIA Stage Instructions

M404 E 06.12.NF.1 AZ-STD Diascopic Stand and AZ-STGD DIA Stage Instructions

Introduction Thank you for purchasing the Nikon products. This instruction manual is written for the users of the Nikon AZ-STD Diascopic Stand and AZ-STGD DIA Stage. To ensure correct usage, read this manual carefully before operating the product. It is prohibited to reproduce or transmit this manual in part or whole without Nikon s expressed permission. The contents of this manual are subject to change without notice. Although every effort has been made to ensure the accuracy of this manual, if you note any points that are unclear or incorrect, contact your nearest Nikon representative. Some of the products described in this manual may not be included in the set you have purchased. Also be sure to read the manuals for any other products that you are using with this product. If the equipment is used in a manner not specified by the manufacturer, the protection provided by the equipment may be impaired. 1

Safety Precautions To ensure correct and safe operation, read this manual before using the products. Warning/Caution Symbols Used in This Manual Although Nikon products are designed to provide you with the utmost safety during use, incorrect usage or disregard of the instructions can cause personal injury or property damage and will lead to the forfeiture of all claims against warranty. For your safety, read the instruction manual carefully and thoroughly before using the products. And, do not discard this manual but keep it near the products for easy reference. Safety instructions within this manual are accompanied by the following symbols to highlight their importance. Be sure to follow the instructions indicated with these symbols. Symbol Warning Caution Details Disregarding instructions marked with this symbol may lead to death or serious injury. Disregarding instructions accompanying this symbol may lead to injury or property damage. 2

Safety Precautions Warning 1. Intended application of the products The products must only be used for microscopic observation. Do not use the products for other purpose. 2. Do not disassemble. Disassembling the products may result in malfunctions. Malfunctions and damage due to disassembling are unwarranted. Do not disassemble the section that is not indicated in this manual. If you notice any abnormalities, contact your nearest Nikon representative. 3. Read the instruction manual carefully. To ensure safety, carefully read this manual and the manuals for other equipment used with the products. Especially be sure to follow the warnings and cautions indicated in the first of this manual. 4. Use the specified light source. Use the products with the following light sources. Do not use the products with the devices other than the following light sources. Nikon C-FI115D Fiber Illuminator (for Japan) Nikon C-FI115 Fiber Illuminator (for the countries excluding Japan where the power supply is 100 to 120 VAC) Nikon C-FI230 Fiber Illuminator (for the countries where the power supply is 230 VAC) 5. Check the input voltage. To use a light source, check the input voltage indicated on its nameplate before turning on the light source. If the input voltage indicated on the places differs from the local voltage, do not turn on the light source. Contact your nearest Nikon representative. Use of a product under an incorrect voltage may cause over-heating due to over-current, possibly resulting in fire and damaging the light sources. 6. Use the specified lamp and power cord. Be sure to use the lamp and power cord specified in the instruction manual. Use of other devices may cause malfunctions or fire. 7. Use the provided light guide fiber. Be sure to use the light guide fiber provided with these products. Use of other devices may cause malfunctions or fire. 8. Pay attention for heat from the light source. The lamp and its surrounding areas become very hot during and immediately after illumination. Follow the cautions below to avoid burn injury and to prevent fire. Do not touch the lamp or its surroundings while the lamp is lit or for 30 minutes after turning off the power. Be sure to use the product with the top cover of the light source closed. Cool down the lamp and its surrounding areas (for approximately 30 minutes after being turned off) before replacing the lamp. Do not allow cloth, paper, or highly flammable volatile materials, such as gasoline, benzine, paint thinner or alcohol, to come near the light source while the lamp is lit or within 30 minutes after turning off the power. Do not block the opening on the side of the light source and air vents on the back. If any obstacle covers the light source or something is placed on the light source, the heat dissipation is inhibited and the light source becomes abnormally hot. 3

Safety Precautions Caution 1. Turn off the power of the light source before assembly or maintenance. To prevent electric shocks and fire, be sure to turn off the power of the light source by pressing down the power switch to O side and unplug the power cord when assembling the product, attaching the light guide fiber, replacing lamps or filters, or performing maintenance such as cleaning of the objective. Also, be sure to turn off the light source by pressing down the power switch to the O side before plugging or unplugging the power cord. 2. Do not wet the product nor allow foreign matter to be intruded. Do not wet the product. Water splashed onto any component of the products may result in malfunction. Also, malfunctions may occur if foreign matters intrude. If any liquid or foreign matters intrude, do not use the product, but contact your nearest Nikon representative. 4

Safety Precautions Notes on Handling the System 1. Carefully handle the products. These products are optical and precision instruments. Handle them carefully and do not subject them to damage or vibrations. Cautions on moving the microscope system Securely hold the bottom front of the microscope base and the indentations at the bottom back of the stage pillar when carrying the microscope. Do not grasp the stand pillar, elevating section, stage, stage height adjustment knob, focus knob, eyepiece tube, zoom body, etc. when carrying the microscope. If grasped, these parts may be disconnected inadvertently, and more, malfunctions or adverse effects on their performance may occur. 2. Installation or storage location These products are optical and precision instruments. Therefore, using or storing the products under unsuitable conditions may damage them or may have an adverse effect on their performance. The following conditions should be considered when selecting the installation or storage location. For installation, select a location with a temperature between 0 to 40 C and a relative humidity of 85 % or less (no condensation). For storage, select a location with a temperature between -20 to 60 C and a relative humidity of 90% or less (no condensation). If the microscope system is installed or stored in a location subject to high temperatures and humidity, mold or condensation may form on a lens, a malfunction may occur, or the performance may deteriorate. Avoid bright locations where the products are exposed to direct sunlight or immediately under room lights. The image quality deteriorates in bright surroundings because of the ambient light entering the objective. Install in a place with little dust and dirt. Install in a place with little vibrations. Install and store the products on a sturdy desk or table that can bear the weight of the product. Install the products in the location of 10 cm or more away from the surrounding walls. Do not install in the narrow space such as shelf or locker. Do not place anything on the products. Cover the products to avoid dust when storing. Arrange a layout that allows easy removal of the power cord from the AC inlet of the light source in the event of an emergency. 3. Weak electromagnetic waves The light source emits weak electromagnetic waves. Do not install the light source near precision electronic devices to avoid degrading their performance. If a TV or radio reception is affected, move the TV or radio set farther from the light source. 4. Notes on handling optical parts Scratch or dirt such as fingerprints on an optical part such as a lens or filter affects the viewing of the image. Carefully handle the optical parts. If any dirt attaches to the optical parts, clean them according to Chapter 6, Care and Maintenance. 5. Handling the light guide fiber Do not forcefully bend, pull or twist the light guide fiber. The inlet and outlet attachments are provided to the both ends of the light guide fiber. Make sure that there is no dirt or dust on the light guide fiber. If any dirt attaches to the fiber, clean the fiber according to Chapter 6, Care and Maintenance. 5

Contents Introduction...1 Safety Precautions...2 Warning/Caution Symbols Used in This Manual... 2 Notes on Handling the System... 5 Chapter 1 Part Names...7 Chapter 2 Microscopy...8 Standard Microscopy... 8 Bright-Field Microscopy under the Diascopic Illumination... 11 Simplified Polarization Microscopy under the Diascopic Illumination... 14 DIC Microscopy under the Diascopic Illumination... 16 Oblique Illumination Microscopy...19 Chapter 3 Operation Procedures...20 Operating the Diascopic Illumination... 20 Opening/Closing the Diascopic Illumination Shutter... 21 Operating the Filters... 21 Setting the Specimen and Adjusting the Stage... 21 Focusing on the Specimen... 22 Operating the Condenser... 24 Operating the Field Diaphragm (for the High Magnification Objective)... 26 Operating the Aperture Diaphragm... 27 Operating the Sliders... 28 Operating the Rotatable Polarizer... 28 Operating the Analyzer... 29 Operating the DIA Lambda Plate... 29 Operating the DIC Prism... 30 Operating the OI Slider... 32 Chapter 4 Assembly...33 Installation of the Diascopic Stand... 34 Attaching the Dia Stage and the Stage Glass... 35 Microscope Assembly... 36 Connecting the Light Guide Fiber... 37 Attaching the Sliders... 38 Chapter 5 Troubleshooting...39 Troubleshooting on Image Viewing... 39 Troubleshooting on the Light Source... 41 Chapter 6 Care and Maintenance...42 Cleaning the Lenses, the Filters, and the Light Guide Fiber... 42 Cleaning the Product... 42 Decontamination... 42 Storage of the Product... 42 Regular Inspections (Charged)...42 Chapter 7 Specifications...43 AZ-STD Diascopic Stand... 43 AZ-STGD DIA Stage... 44 6

1 Part Names The Nikon AZ-STD Diascopic Stand is the stand for diascopic illumination exclusive to the AZ series microscope. The AZ-STD Diascopic Stand is used for the microscopy under the diascopic illumination with the optional AZ-STGD DIA Stage and the C-FI115D/C-FI115/C-FI230 Fiber Illuminator. Configuration example: the figure below shows the configuration example of the following parts. AZ-TE80 Ergonomic Trinocular Tube 80, AZ-W10X Eyepiece 10X, AZ100 Zoom Body, AZ-FM AZ (Focusing Mount) Adapter, AZ-NP3 Triple Nosepiece, objectives, AZ-STD Diascopic Stand (light guide fiber attached), AZ-STGD DIA Stage, AZ-SG Stage Glass, and C-FI115D/C-FI115/C-FI230 Fiber Illuminator. * Additionally, filters are also attached for the DIC microscopy under the diascopic illumination. Ergonomic trinocular tube (AZ-TE80 / AZ-TE100) AZ (Focusing mount) adapter (AZ-FM) Elevating section Fiber illuminator DIA stage and stage glass (AZ-STGD and AZ-SG) (C-FI115D / C-FI115 / C-FI230) (for diascopic illumination) Zoom body (AZ100) Objective Pillar Focus knob (both sides) Triple nosepiece (AZ-NP3) Filter selector lever Diaphragm control thumbwheel Light guide fiber (provided with the AZ-STD) Stage height adjustment knobs (both sides) Diascopic stand (AZ-STD) Stage movement knob High magnification condenser aperture diaphragm open/close lever High magnification condenser centering screw (both sides) Condenser selector knob DIA Stand left view Diascopic illumination shutter open/close lever High magnification condenser centering screw (both sides) High magnification condenser focus knob (on the rear) Slots for filters - Diascopic polarizer - Lambda plate - Low magnification DIC prism / OI silder Slots for the high magnification DIC prism / OI silder 7

2 Microscopy Standard Microscopy ~ Preparation for microscopy ~ This section indicates the standard procedure for microscopy under the diascopic illumination. 1 2 3 4 Switch the optical path of the eyepiece tube to the binocular part. The optical path is switched to the binocular part by pressing the optical path selector lever on the eyepiece tube. Open the diascopic illumination shutter. Open the diascopic illumination shutter by pulling forward the diascopic illumination shutter open/close lever on the left of the DIA stage toward you. Set a specimen. Set a specimen on the stage. Use an appropriate plate or holder according to the shape of the specimen. Turn on the diascopic illumination lamp by turning on the fiber illuminator for the diascopic illumination. For details on operating the illuminator, refer to the instruction manual for the illuminator. 5 Adjust the brightness of the diascopic illumination. Adjust the brightness of the diascopic illumination properly with the brightness control knob on the optical fiber light source and the ND filter on the diascopic stand. To adjust the illumination to a daylight color, set the brightness control knob on the optical fiber light source to the position shown in the right figure, and then, insert/extract the ND filters on the diascopic stand. 45 o MIN MAX Dial position for a daylight color 6 7 8 9 Place the desired objective into the optical path (only when using the triple nosepiece). When the triple nosepiece is used, objectives are selectable. Switch the condenser according to the objective in use. Two condenser lenses are provided, one for low magnification objectives (0.5x to 1x) and another for high magnification objectives (2x to 5x). There are index marks on the right of the condenser selector knob at the microscope base for the knob position. The index marks show the magnifications (0.5 to 1x, or 2 to 5x) and the color codes of objectives. Select the suitable condenser for the objective to be used. Focus on the specimen by adjusting the position of the specimen. Move the specimen so that the position to be observed comes to the center of the viewfield. Focus on the specimen by rotating the focus knobs and the stage height adjustment knobs. The index mark on the right of the stage elevating section indicates the vertical position of the stage. When the mark is positioned at the top of the microscope base, the stage is positioned at the center of the movable range (reference position). Adjust the interpupillary of the binocular part and the diopter of the eyepieces. For details, refer to the instruction manual for the AZ100 Zoom Body. 8

Chapter 2 Microscopy 10 11 Center and focus the condenser (only when using the condenser for the high magnification objective). Center the condenser by rotating the condenser centering screws on the right and left of the microscope and rotate the condenser focus knob on the right to form the field diaphragm image on the specimen surface. For details, refer to Operating the Condenser on page 24. Adjust the diaphragms Usually the diaphragms of the AZ100 are used with fully opened because the actual size of the viewfield and the numerical aperture will change with the zoom magnification adjustment. But, when the zoom magnification is fixed and fine adjustments are required, the following operations can be performed. Adjust the field diaphragm. (only for the objectives of 2x to 5x) The field diaphragm restricts the illumination to the area on the specimen to be observed. Adjust the field diaphragm by rotating the diaphragm control thumbwheel on the right of the diascopic stand until the field diaphragm image circumscribes the viewfield. (See page 26.) Adjust the aperture diaphragm. The aperture diaphragm controls the numerical aperture of the illumination system. It is closely related to the resolution of the optical image, contrast, and depth of focus. The aperture diaphragm must be controlled with one of two parts depending on the magnification of the objective. For the high magnification objective (2x to 5x): Rotate the aperture diaphragm open/close lever on the lower part of the high magnification condenser lens to adjust the aperture diaphragm to 70 to 80% of the numerical aperture of the observation optical system. (See page 27.) For the low magnification objective (0.5x or 1x): Rotate the diaphragm control thumbwheel on the right of the diascopic stand to adjust the aperture diaphragm to 70 to 80% of the numerical aperture of the observation optical system. (See page 27.) 12 13 Observe the specimen with the zoom operation. The magnification can be adjusted in the range of 1x to 8x of the selected objective with the zoom knob. The specimen can be observed according to the following microscopies when the diascopic illumination is used. Bright-field microscopy under the diascopic illumination (See page 11.) Simplified polarization microscopy under the diascopic illumination (See page 14.) DIC microscopy under the diascopic illumination (See page 16.) Oblique Illumination microscopy (See page 19.) Turn off the lamp after observations. Turn off the illuminator for the lamp after observations. When covering the microscope or the illuminator, wait until the illuminator cools sufficiently. 9

Chapter 2 Microscopy Standard procedure 9 Adjust the interpulillary and the diopter. Use the optical path for the binocular part. 1 12 Observe the specimen with the zoom operation. 6 Select the desired magnification. 3 Set the specimen. 2 4 Open the diascopic illumination shutter. Turn on the diascopic illumination lamp. 5 Adjust the brightness of the diascopic illumination. 13 Turn off the lamp after observations. Focus on the specimen. Adjust the condenser according to the objective. Center and focus the condenser. Adjust the aperture diaphragm. (for the high magnification objective) Adjust the brightness. Adjust the diaphragm. (field diaphragm for high magnifications, aperture diaphragm for low magnifications) Focus on the specimen by adjusting the position of the specimen. 8 7 10 11 5 11 8 10

Chapter 2 Microscopy Bright-Field Microscopy under the Diascopic Illumination This section indicates the procedure for the bright-field microscopy under the diascopic illumination. 1 2 Prepare for operation. Prepare the microscope and a specimen according to the steps 1 to 11 of Standard Microscopy. (See pages 8 to 11.) Remove the polarizer, analyzer, DIC prism, lambda plate, and the OI slider from the optical path. When these sliders are attached, remove them from the optical path by pulling them to the click-stop position. 3 Adjust the brightness of the diascopic illumination. Adjust the brightness of the diascopic illumination properly with the brightness control knob on the illuminator and the ND filter on the diascopic stand. To adjust the illumination to a daylight color, set the brightness control knob on the optical fiber light source to the position shown in the right figure, and then, insert/extract the ND filters on the diascopic stand. 45 o MIN MAX Dial position for a daylight color 4 5 6 7 Place the desired objective into the optical path (only when using the triple nosepiece). When the triple nosepiece is used, objectives are selectable. Select the condenser according to the objective in use. Two condenser lenses are provided, one for low magnification objectives (0.5x or 1x) and another for high magnification objectives (2x to 5x). There are index marks on the right of the condenser selector knob at the microscope base for the knob position. The index marks show the magnifications (0.5 to 1x, or 2 to 5x) and the color codes of objectives. Select the suitable condenser for the objective to be used. Set the zoom magnification properly. The zoom operation is enabled for the selected objective in the range of 1x to 8x by rotating the zoom knob. Focus on the specimen by adjusting the position of the specimen. Move the stage to adjust the position of the specimen so that the observation point is located at the center of the viewfield, and then, focus on the specimen with the focus knobs and the stage height adjustment knobs. 11

Chapter 2 Microscopy 8 Adjust the diaphragms Usually the diaphragms of the AZ100 are used with fully opened because the actual size of the viewfield and the numerical aperture will change with the zoom magnification adjustment. But, when the zoom magnification is fixed and fine adjustments are required, the following operations can be performed. Adjust the field diaphragm. (only for the objectives of 2x to 5x) The field diaphragm restricts the illumination to the area on the specimen to be observed. Adjust the field diaphragm by rotating the diaphragm control thumbwheel on the right of the diascopic stand until the field diaphragm image circumscribes the viewfield. (See page 26.) Adjust the aperture diaphragm. The aperture diaphragm controls the numerical aperture of the illumination system. It is closely related to the resolution of the optical image, contrast, and depth of focus. The aperture diaphragm must be controlled with one of two parts depending on the magnification of the objective. For the high magnification objective (2x to 5x): Rotate the aperture diaphragm open/close lever on the lower part of the high magnification condenser lens to adjust the aperture diaphragm to 70 to 80% of the numerical aperture of the observation optical system. (See page 27.) For the low magnification objective (0.5x or 1x): Rotate the diaphragm control thumbwheel on the right of the diascopic stand to adjust the aperture diaphragm to 70 to 80% of the numerical aperture of the observation optical system. (See page 27.) 9 Observe the specimen. 12

Chapter 2 Microscopy Bright-field microscopy under the diascopic Illumination 1 Prepare for operation (See pages 8 to 10.) 9 Observe the specimen. 6 Set the zoom magnification properly. 2 Remove the sliders from the optical path. 4 Locate the desired objective in place. 3 Adjust the brightness of the diascopic illumination. Focus on the specimen. Select the condenser according to the objective. Adjsut the aperture diaphragm. (for high magnification objective) Adjust the brightness of the diascopic illumination. Adjust the diaphragm. (field diaphragm for high magnifications, aperture diaphragm for low magnifications) Adjust the position of the specimen and fucus on the specimen. 7 5 8 3 8 7 Left view of the DIA stand 2 Remove the sliders from the optical path 13

Chapter 2 Microscopy Simplified Polarization Microscopy under the Diascopic Illumination This section indicates the procedure for the simplified polarization microscopy under the diascopic illumination. Necessary accessories The following accessories are necessary for the simplified polarization microscopy under the diascopic illumination. AZ-RP Rotatable Polarizer AZ-AN DIA DIC Prism Holder with Analyzer 1 2 3 4 Focus on a specimen with the bright-field microscopy under the diascopic illumination. Focus on a specimen with the bright-field microscopy under the diascopic illumination according to Bright-Field Microscopy under the Diascopic Illumination. Place the rotatable polarizer and the analyzer into the optical path. Insert the rotatable polarizer into the polarizer slot on the left of the diascopic stand. When the rotatable polarizer is inserted as far as it goes, it is placed into the optical path. And when the rotatable polarizer is pulled to the click-stop position, it is removed from the optical path. Insert the analyzer into the slot on the front right of the zoom body. When the analyzer is inserted as far as it goes, it is placed into the optical path. And when the analyzer is pulled to the click-stop position, it is removed from the optical path. Adjust the orientation of the polarizer and observe the specimen. Adjust the orientation of the polarizer by rotating the diascopic polarizer thumbwheel. The viewing of the image is changed by adjusting the orientation of the polarizer. There are mark on the polarizer thumbwheel. When the polarizer thumbwheel is aligned with mark, the orientation of the polarizer is positioned in the horizontal direction, which is at the right angle to the orientation of the analyzer (crossed Nicols position). When the orientations of the polarizer and the analyzer are crossed Nicols position, the view becomes darkest. For details on the rotatable polarizer, refer to Operating the Rotatable Polarizer on page 28. Adjust the brightness if necessary. If the brightness is not enough at the crossed Nicols position, adjust the brightness of the diascopic illumination. 14

Chapter 2 Microscopy Simplified polarization microscopy under the diascopic illumination 1 Focus on the specimen with the bright-field microscopy under the diascopic illumination. (See page 11 to 13.) 3 Observe the specimen. Left view of the diascopic stand 2 Place the analyzer into the optical path. Place the polarizer into the optical path. Adjust the orientation of the polarizer. 2 3 4 Adjust the brightness if necessary. Adjust the brightness if necessary. 4 15

Chapter 2 Microscopy DIC Microscopy under the Diascopic Illumination This section indicates the procedure for the differential interference contrast (DIC) microscopy under the diascopic illumination. Necessary accessories The following accessories are necessary for the simplified polarization microscopy under the diascopic illumination. AZ-RP Rotatable Polarizer AZ-AN DIA DIC Prism Holder with Analyzer AZ-DP1 / AZ-DP4 / AZ-DP5 DIA DIC Prism AZ-DPS1 / AZ-DPS5 DIA DIC Prism Slider AZ-DL DIA DIC Lambda Plate (for the sensitive color DIC microscopy) AZ-Plan Apo 1x / AZ-Plan Apo 4x / AZ-Plan Fluor 5x objective (The DIC microscopy cannot be performed with other objectives.) 1 2 3 4 Focus on a specimen with the bright-field microscopy under the diascopic illumination. Focus on a specimen with the bright-field microscopy under the diascopic illumination according to Bright-Field Microscopy under the Diascopic Illumination. Place the rotatable polarizer and the analyzer into the optical path. Insert the rotatable polarizer into the polarizer slot on the left of the diascopic stand. When the rotatable polarizer is inserted as far as it goes, it is placed into the optical path. And when the rotatable polarizer is pulled to the click-stop position, it is removed from the optical path. Insert the analyzer into the slot on the front right of the zoom body. When the analyzer is inserted as far as it goes, it is placed into the optical path. And when the analyzer is pulled to the click-stop position, it is removed from the optical path. Adjust the orientation of the polarizer to be at the crossed Nicols position. Rotate the diascopic polarizer thumbwheel so that the view becomes darkest. There are mark on the polarizer thumbwheel. When the polarizer thumbwheel is aligned with mark, the orientation of the polarizer is positioned in the horizontal direction, which is at the right angle to the orientation of the analyzer (crossed Nicols position). When the orientations of the polarizer and the analyzer is crossed Nicols position, the view becomes darkest. For details on the rotatable polarizer, refer to Operating the Rotatable Polarizer. Adjust the brightness if necessary. If the brightness is not enough at the crossed Nicols position, adjust the brightness of the diascopic illumination. 16

Chapter 2 Microscopy 5 6 7 8 9 Place the DIC prism (DIA DIC prism 1x / 4x / 5x) on the illumination side into the optical path. The prism to be used and its inserting position change depending on the magnification of the objective. The color code of the objective is displayed on the DIA DIC prism and the slot. Insert the DIA DIC prism into an appropriate slot according to the color code of the objective. For the 1x objective: insert the AZ-DP1 DIA DIC Prism 1x into the slot for the DIC or OI slider on the left rear of the diascopic stand. For the 4x objective: insert the AZ-DP4 DIA DIC Prism 4x into the slot for the high magnification condenser on the left of the microscope. For the 5x objective: insert the AZ-DP5 DIA DIC Prism 5x into the slot for the high magnification condenser on the left of the microscope. When the slider is inserted as far as it goes, the prism is placed into the optical path. And when the slider is pulled to the click-stop position, the prism is removed from the optical path. Place the DIC prism on the objective side (DIA DIC prism slider 1-4x / 5x) into the optical path. Insert the DIA DIC prism slider into the slot on the analyzer on the AZ100 Zoom Body. An appropriate slider differs depending on the magnification of the objective. For the 1x/4x objective: insert the AZ-DPS1 DIA DIC Prism Slider (1-4x). For the 5x objective: insert the AZ-DPS5 DIA DIC Prism Slider (5x). Loosen the clamp screw on the side of the analyzer. Insert the DIA DIC prism slider as far as it goes to place the prism into the optical path and pull the DIA DIC prism slider fully to remove the prism from the optical path. Be careful that the DIA DIC prism slider is detached if the clamp screw is loosen too much. Select the interference color. Set an interference color by rotating the prism position knob at the end of the DIA DIC prism slider. Place the lambda plate into the optical path if necessary. When the lambda plate is placed into the optical path in a condition that the interference color is gray, the interference color changes to sensitive red. This color is useful for the high color contrast observation. Insert the AZ-DL DIA DIC Lambda Plate into the slot for the lambda plate on the left rear of the diascopic stand. When the lambda plate is inserted as far as it goes, it is placed into the optical path. And when the lambda plate is pulled to the click-stop position, it is removed from the optical path. Observe the specimen. 17

Chapter 2 Microscopy Procedure for the DIC microscopy under the diascopic illumination 1 Focus on the specimen with the bright-field microscopy under the diascopic illumination. (See pages 11 to 13.) 9 Observe the specimen. 2 Place the analyzer into the optical path. 6 Place the DIA DIC prism into the optical path. 7 4 Left view of the diascopic stand (forthe low magnification objectives) Select the interference color. Adjust the brightness if necessary. Place the polarizer into the optical path. 2 Adjust the orientation of the polarizer to be at the crossed Nicols position. 3 Place the DIC prism into the optical path. 5 Place the lambda plate into the optical path. 8 Left view of the diascopic stand (for the high magnification objectives) 2 3 Place the polarizer into the optical path. 8 Place the lambda plate into the optical path. Adjust the orientation of the polarizer to be at the crossed Nicols position. 18 Place the DIC prism into the optical path. 5

Chapter 2 Microscopy Oblique Illumination Microscopy This section indicates the procedure for the oblique illumination microscopy with the OI slider. Oblique illumination The oblique illumination microscopy is performed as follows: A part of the illumination is shielded with the sliding diaphragm. Coherent light is projected obliquely onto the specimen. The specimen is shaded and its contrast increases. Necessary accessories The following accessory is necessary for the oblique illumination microscopy. AZ-OI OI Slider 1 2 3 4 5 Focus on a specimen with the bright-field microscopy under the diascopic illumination. Focus on a specimen with the bright-field microscopy under the diascopic illumination according to Bright-Field Microscopy under the Diascopic Illumination on pages 11 to 13. Place the OI slider into the optical path. The inserting position for the slider changes depending on the magnification of the objective. The color code of the objective is displayed on the slot. Insert the slider into an appropriate slot according to the color code of the objective. For the 0.5x or 1x objective: insert the slider into the slot for the DIC/OI slider on the left rear of the diascopic stand. For the 2x to 5x objective: insert the slider into the slot for the high magnification condenser on the left of the microscope. Adjust the contrast with the knob on the OI slider. Adjust the contrast of the image by moving the sliding diaphragm right and left with the knob on the OI slider. To open the diaphragm fully, align the index on the knob with the click-stop position. Adjust the brightness if necessary. If the brightness is less than the desired brightness when the sliding diaphragm is in the optical path, adjust the brightness of the diascopic illumination. Observe the specimen. 19

3 Operation Procedures This chapter indicates the details on operating this product. Color code For this product, according to the magnification of the objective, switch the condenser lens and select an appropriate slider and insert it into a proper slot. The color code of the objective is displayed on such part that must be switched according to the magnification of the objective. For the color code of the objective, see the right figure. Objective AZ-Plan Apo 0.5x AZ-Plan Apo 1x AZ-Plan Fluor 2x AZ-Plan Apo 4x AZ-Plan Fluor 5x Color code Silver Gold White Red Orange Operating the Diascopic Illumination Turning on/off the lamp Turning on/off the power supply of the optical fiber light source to turn on/off the lamp. Press down the power switch to the I side to turn on the lamp, and to the O side to turn off. Brightness control The brightness of the illumination is adjusted with the brightness control knob on the optical fiber light source. Turn the brightness control knob clockwise to make the illumination brighter, and turn the knob counterclockwise to make the illumination darker. Adjusting the illumination to a daylight color The illumination becomes a daylight color when the brightness control knob on the optical fiber light source is aligned to the position as shown in the right figure. Adjust the brightness by pushing in or pulling out the ND filter on the right of the diascopic stand. Power switch Brightness control knob Operation of the diascopic illumination 45 o MIN MAX For detailed operation of the optical fiber light source, refer to the instruction manual for the optical fiber light source. Knob position for a daylight color 20

Chapter 3 Operation Procedures Opening/Closing the Diascopic Illumination Shutter The shutter for blocking the diascopic illumination is located on the substage of the diascopic stand. When performing a microscopy under the diascopic illumination such as the bright-field microscopy, the simplified polarization microscopy, or the DIC microscopy, open the shutter to enter the diascopic illumination into the optical path. When performing the epi-fl microscopy with the AZ-FL Epi-Fluorescence Attachment, close the shutter not to enter the diascopic illumination into the optical path. Diascopic illumination shutter open/close lever (Far side: closing the shutter, front side: opening the shutter) Opening/closing the diascopic illumination shutter Operating the Filters Three filter selector levers are located on the right of the product. Pull the lever to remove the filter from the optical path and press it as far as it goes to place the filter in the optical path. Filter type Intended application ND2 (ND filter) For brightness control (transmittance: 50%) ND4 (ND filter) For brightness control (transmittance: 25%) ND16 (ND filter) For brightness control (transmittance: 6%) Filter selector lever Operation of the filters The NCB14 filter is equipped with the product. The NCB14 filter cannot be removed from the optical path. Setting the Specimen and Adjusting the Stage Setting the specimen Sufficiently lower the stage by rotating the stage height adjustment knob and place the specimen on the stage without being touched to the objective. Adjusting the stage (fine focus) To change the observation point after setting the specimen on the stage, rotate the stage movement knobs so that the specimen can be finely moved. The upper knob is used to move the stage back and forth and the lower knob is used to move the stage right and left. Stage movement knobs Adjusting the stage (fine focus) 21

Chapter 3 Operation Procedures Focusing on the Specimen Caution Do not rotate the right and left focus knobs and the right and left stage height adjustment knobs in the opposite direction. Do not rotate those knobs forcibly over the limit. Do not rotate the fine focus knob with holding the coarse focus knob nor vice versa. Doing so may cause damage to the product. For this product, focusing on the specimen can be performed in two ways, with the stage height adjustment knobs or the focus knobs. Use either of those knobs that allow you to operate easier and focus on the specimen. Adjusting the zoom body vertically The zoom body is adjusted vertically with the focus knobs on the both sides of the focus elevating section of the pillar. The coarse focus knob and fine focus knob are provided. One revolution of the coarse focus knob allows the stage to move by 18.5 mm and that of the fine focus knob allows by 3.27 mm. Zoom body Focus knobs (both sides) Coarse focus knob Fine focus knob Torque for the focus knob The torque adjustment ring is equipped inside the left focus knob. Adjust the torque enough to prevent the focusing mount from descending with its own weight by inserting the hexagonal screwdriver into a hole on the ring. To increase the torque, rotate the screwdriver counterclockwise. To decrease the torque, rotate the screwdriver clockwise. Adjusting the stage vertically The stage is adjusted vertically with the stage height adjustment knobs on the both sides of the stand. The coarse focus knob and fine focus knob are provided. One revolution of the coarse focus knob allows the stage to move by 37.7 mm and that of the fine focus knob allows by 0.27 mm. The index mark on the right of the stage elevating section indicates the vertical position of the stage. When the mark is positioned at the top of the microscope base, the stage is positioned at the center of the movable range (reference position). See the right figure. Stage Torque Stage height adjustment ring Fine focus knob adjustment knobs (both sides) Coarse focus knob Focusing on the specimen Index mark Index mark on the elevating section of the stage Torque for the stage height adjustment knob The torque adjustment ring is equipped on the left of the stage height adjustment knob. The torque required to rotate the knob can be adjusted by rotating the ring. The torque is increased by rotating the torque adjustment ring in the direction of arrow of TORQUE. Adjust the torque enough to prevent the stage from descending with its own weight. 22

Chapter 3 Operation Procedures Focusing procedure Perform the following operations to focus on the specimen. (1) Check the working distance of the objective. The distance between the focal plane and the lower end of the objective is called working distance, which displayed on the objective as WD. Focusing will be easier by adjusting the distance between the specimen and the objective considering the working distance. (2) Focus on the specimen. Two types of knobs are used for focusing on the specimen. The first type is the focus knobs to move the elevating section of the stand, the second type is the stage height adjustment knobs to move the stage up/down. Use the coarse focus knobs for coarse adjustment and use the fine focus knobs for fine adjustment. To focus on the specimen under a high magnification, the fine focus knobs are recommended. 23

Chapter 3 Operation Procedures Operating the Condenser Selecting From Between the Low Magnification Condenser and the High Magnification Condenser The diascopic stand has the condenser lens for objectives of low magnifications (0.5x to 1x) and another condenser lens for objectives of high magnifications (2x to 5x). Switch those condenser lenses according to the objective in use. Use the condenser selector knob to switch the condenser. Move the knob to the 0.5-1x side (far side) to place the low magnification condenser into the optical path and move it to the 2-5x side (front side) to place the high magnification condenser into the optical path. The color code of the objective is displayed on the condenser selector knob, indicating the position of the condenser. Condenser Objective Color code Condenser selector knob Low magnification 0.5x, 1x Silver and gold 0.5-1x High magnification 2x, 4x, 5x White,red, and orange 2-5x Use of the high magnification condenser for the objective of 2x and the zoom magnification of 1.3x or less may cause vignetting in the viewfield. Use the low magnification condenser to keep the viewfield clear. Low magnification condenser and high magnification condenser 24

Chapter 3 Operation Procedures Focusing and Centering the High Magnification Condenser For use of the high magnification condenser, the condenser must be focused and centered so that the light passing through the condenser forms an image at the correct location (the center of the optical path) on the specimen surface. Once the condenser is adjusted before operation, it does not need farther adjustment. 1. Place a specimen on the stage. 2. Place the high magnification condenser in the optical path. 3. Rotate the zoom knob of the AZ100 to the position of minimum magnification and then operate the focus knob and stage height adjustment knob to focus on the specimen. 4. Rotate the diaphragm control thumbwheel on the right of the diascopic stand to decrease the size of the field diaphragm. 5. Rotate the condenser focus knob to form the field diaphragm image on the specimen surface. Condenser focus knob (on the rear surface of the condenser slider) Condenser centering screws (both sides) Field diaphragm image Eyepiece viewfield Focusing and centering of the high magnification condenser 6. Turn the two condenser centering screws (on both sides) so that the field diaphragm image is positioned in the center of the view field. 7. Set the zoom knob of the AZ100 to 8x and then operate the focus knob and stage height adjustment knob to focus on the specimen. 8. Rotate the condenser focus knob to form the field diaphragm image on the specimen surface. 9. Adjust the diaphragm control thumbwheel and the condenser centering screws so that the field diaphragm image inscribes the view field. 10. To observe the specimen, rotate the field diaphragm adjuster so that the field diaphragm image circumscribes the viewfield. An appropriate size of the field diaphragm differs depending on the zoom magnification. Therefore, the field diaphragm is fully opened in general operation. Once the Zoom magnification is fixed and fine adjustment is required, change the size of the field diaphragm. 25

Chapter 3 Operation Procedures Operating the Field Diaphragm (for the High Magnification Objective) The field diaphragm restricts the illumination to the area on the specimen to be observed. Operate the diaphragm control thumbwheel on the right of the diascopic stand to adjust the size of the field diaphragm for the diascopic illumination. An appropriate size of the field diaphragm differs depending on the zoom magnification. Therefore, the field diaphragm is fully opened in general operation. Once the zoom magnification is fixed and fine adjustment is required, change the size of the field diaphragm. Normally, adjust the size of the diaphragm so that the image of the diaphragm circumscribes the viewfield. If a broader area than necessary is illuminated, stray light may enter the optical system, creating flaring, and reducing the contrast of the optical image. Perform the field diaphragm adjustment after completing focusing and centering of the condenser lens. When the low magnification objective (low magnification condenser) is attached, the field diaphragm cannot be used. Diaphragm control thumbwheel Field diaphragm image Eyepiece viewfield Operation of the field diaphragm 26

Chapter 3 Operation Procedures Operating the Aperture Diaphragm The aperture diaphragm controls the numerical aperture of the illumination system, closely related to the resolution of the optical image, contrast, and depth of focus. For the high magnification condenser When the high magnification condenser is attached, use the aperture diaphragm adjuster on the bottom of the condenser to adjust the aperture diaphragm. To adjust the size of the aperture diaphragm, rotate the aperture diaphragm adjuster. Remove one of the eyepieces, and then adjust the aperture diaphragm to about 70% to 80% of the pupil of the objective while observing the pupil of the objective in the eyepiece tube. (Refer to the right figure.) Aperture diaphragm adjuster For high magnification condenser Perform the aperture diaphragm adjustment after completing focusing and centering the condenser lens. An appropriate size of the aperture diaphragm differs depending on the zoom magnification. Usually, fully open the aperture diaphragm. When the observation magnification is determined, adjust the size. For the low magnification condenser When the low magnification condenser is attached, use the diaphragm control thumbwheel on the right of the diascopic stand to adjust the aperture diaphragm. Remove one of the eyepieces, and then adjust the aperture diaphragm to about 70% to 80% of the pupil of the objective while observing the pupil of the objective in the eyepiece tube. (Refer to the right figure.) Diaphragm control thumbwheel For low magnification condenser Pupil of the objective 70~80 100 Aperture diaphragm Operating the aperture diaphragm 27

Chapter 3 Operation Procedures Operating the Sliders Various slots are provided on the left of the diascopic stand to insert sliders according to the microscopy. This section indicates the procedure for operating each slider. Operating the Rotatable Polarizer Usage of the rotatable polarizer The simplified polarization microscopy under the diascopic illumination is enabled in combination with the rotatable polarizer and the analyzer. The DIC microscopy under the diascopic illumination is enabled by using the rotatable polarizer and the analyzer together and inserting a DIC prism into each slot on the illumination side and the objective side. The sensitive color microscopy is enabled when the DIA lambda plate is used under the simplified polarization microscopy under the diascopic illumination or the DIC microscopy under the diascopic illumination. Placing into or removing from the optical path Insert the rotatable polarizer into the P slot on the left of the diascopic stand with its orientation indication facing toward the front side (observer side). When the rotatable polarizer is inserted as far as it goes, it is placed into the optical path. And when the rotatable polarizer is pulled to the click-stop position, it is removed from the optical path. Adjusting the orientation The orientation of the polarizer and the viewing of the image are changed by rotating the thumbwheel on the rotatable polarizer. Perform the following to make the polarizer and the analyzer at the crossed Nicols position. 1. Place the polarizer and analyzer into the optical path. Insert the analyzer into the slot on the front of the AZ100 Zoom Body. (Refer to Operating the Analyzer. ) 2. When the high magnification condenser is used, maximize the size of the aperture diaphragm. 3. Remove an eyepiece from the eyepiece tube and look into the tube, and then make sure that the shape of the pupil of the objective is a bright circle. 4. Rotate the polarizer thumbwheel until the view becomes darkest. This is the crossed Nicols position. The polarizer and the analyzer are at the crossed Nicols position when the thumbwheel is aligned with mark. A dark cross image appears in the pupil as shown in the right figure. Rotatable polarizer Polarizer thumbwheel Operating the rotatable polarizer Dark cross 28

Chapter 3 Operation Procedures Operating the Analyzer Operating the analyzer The simplified polarization microscopy under the diascopic illumination is enabled in combination with the rotatable polarizer and the analyzer. The DIC microscopy under the diascopic illumination is enabled by using the rotatable polarizer and the analyzer together and inserting a DIC prism into each slot on the illumination side and the objective side. The sensitive color microscopy is enabled when the DIA lambda plate is used under the simplified polarization microscopy under the diascopic illumination or the DIC microscopy under the diascopic illumination. Analyzer Inserting the analyzer Placing into or removing from the optical path Insert the analyzer into the filter slot on the front of the zoom body. When the analyzer is inserted as far as it goes, it is placed into the optical path. And when the analyzer is pulled to the click-stop position, it is removed from the optical path. The analyzer has a slot for inserting the DIC prism slider. Operating the DIA Lambda Plate Operating the DIA lambda plate The sensitive color polarization microscopy under the diascopic illumination is enabled by adding the DIA lambda plate to the condition of the simplified polarization microscopy under the diascopic illumination. The DIC microscopy with the sensitive color is enabled by adding the diascopic lambda plate to the condition of the DIC microscopy under the diascopic illumination. Placing into or removing from the optical path Insert the DIA lambda plate into the λ slot on the left of the diascopic stand. When the DIA lambda plate is inserted as far as it goes, it is placed into the optical path. And when the DIA lambda plate is pulled to the click-stop position, it is removed from the optical path. DIA lambda plate Operating the DIA lambda plate 29

Chapter 3 Operation Procedures Operating the DIC Prism Notice For the DIC microscopy, optical distortion in the vessel, glass slide, and cover glass must be small. Therefore, do not use these parts whose plastic part enters the optical path. Combination of the objective for the DIC microscopy and the DIC prism The color code of an appropriate objective is displayed on the sliders for the DIC prism. Check the color code displayed on the objective and place the DIC prism slider marked with the same color code into the optical path. Objective * 1 AZ-Plan Apo 1x AZ-Plan Apo 4x AZ-Plan Fluor 5x Color code Gold Red Orange DIC prism on the illuminator side * 2 * 3 (Diascopic stand base) AZ-DP1 DIA DIC Prism 1x AZ-DP1 DIA DIC Prism 4x AZ-DP5 DIA DIC Prism 5x DIC prism on the observer side * 3 (AZ100 zoom body) AZ-DPS1 DIA DIC Prism Slider 1-4x AZ-DPS5 DIA DIC Prism Slider 5x *1: The DIC microscopy cannot be performed when the 0.5x or the 2x objective is used. *2: The magnification of an appropriate objective is displayed on the DIA DIC prism. *3: Shear of the DIC prism is optimized for the high zoom magnification. Usage of the DIC prism The DIC microscopy under the diascopic illumination is enabled in a condition that the polarizer and the analyzer are at the crossed Nicols position and the DIC prism is used. The DIC microscopy under the sensitive color is enabled in combination with the DIC lambda plate and other parts required for the DIC microscopy under the diascopic illumination. 30

Chapter 3 Operation Procedures Placing into or removing from the optical path (illuminator side) The inserting position of the DIC prism on the illuminator side differs depending on the objective or the condenser in use. For the 1x objective: insert the AZ-DP1 DIA DIC Prism 1x into the DIC/OI slot on the left of the diascopic stand. For the 4x objective: insert the AZ-DP4 DIA DIC Prism 4x into a slot on the lower part of the high magnification condenser. For the 5x objective: insert the AZ-DP5 DIA DIC Prism 5x into a slot on the lower part of the high magnification condenser. When the DIC prism is inserted as far as it goes, it is placed into the optical path. And when the DIC prism is pulled to the click-stop position, it is removed from the optical path. AZ-DP1 DIA DIC Prism Attaching the DIC prism for low magnification to the illuminator side AZ-DP4 DIA DIC Prism 4x or AZ-DP5 DIA DIC Prism 5x Attaching the DIC prism for high magnification to the illuminator side 31

Chapter 3 Operation Procedures Placing into or removing from the optical path and operating the prism (observer side) To attach the DIC prism to the observer side, the AZ-AN DIA DIC Prism Holder with Analyzer must be attached to the AZ100 Zoom Body. Insert the AZ-DPS1/AZ-DPS5 DIA DIC Slider into the specified slot on the DIA analyzer. Use the AZ-DPS1 DIA DIC Slider for the low magnification condenser and use the AZ-DPS5 DIA DIC Slider for the high magnification condenser. When the DIA DIC prism slider is inserted as far as it goes, the DIC prism is placed into the optical path. And when the analyzer is pulled to the click-stop position, it is removed from the optical path. The DIC prism is moved by rotating the prism movement knob on the tip of the DIA DIC prism slider so that the interference color for the DIC microscopy can be adjusted. AZ100 Zoom Body DIA analyzer DIA DIC prism slider Prism movement knob Attaching the DIA DIC prism slider When the AZ-FL Epi-Fluorescence Attachment is attached to the microscope, the DIC microscopy under the diascopic illumination can be performed by attaching the AZ-FLDIC FL-DIC Prism Holder between the AZ-FL Epi-Fluorescence Attachment and the nosepiece. For details, refer to the instruction manual for the AZ-FL Epi-Fluorescence Attachment. DIA DIC prism slider Operating the OI Slider The OI slider is use to perform the oblique illumination microscopy. The oblique illumination microscopy enables observation with an appropriate contrast by shielding a part of the light beam with the sliding diaphragm and projecting coherent light obliquely onto a transparent specimen for shading. Placing into or removing from the optical path The inserting position of the OI slider differs depending on the magnification of the objective. For the 0.5x / 1x objective: insert the OI slider into the DIC / OI slot on the left of the diascopic stand. For the 2x / 4x / 5x objective: insert the OI slider into the slot on the lower part of the high magnification condenser. Insert the slider as far as it goes. Operating the sliding diaphragm Rotate the knob on the OI slider to adjust the opening of the sliding diaphragm and control the contrast of the image. When the knob is rotated to the click-stop position (reference position), the sliding diaphragm opens fully. 32