Operating Manual for the Olympus BX51 and Image Capture System
Brightfield (transmitted light)………..…………………………..… 2 Brightfield (reflected light)………..…………………………..…….3 Fluorescence……………………………………………………….. 4
Diagram of the BX51 Microscope…………………………………. 5 Guide to switches, filters, knobs, etc………………………………..6
Image acquisition and measurements……………………………….8
Troubleshooting……………………………………………………. 10
BRIGHTFIELD (transmitted light)
Illumination transmits through a transparent sample from the bottom of the microscope.
1. Turn on the halogen light switch (1) for transmitted light to “│” (ON). 2. Engage the brightfield mirror unit (BF) (17).
3. Adjust the light path. The light path selector knob (2) should be pushed in all the way. 4. Select transmitted light (lower position) on the switch by the brightness indicator (4). 5. Adjust the light intensity using the brightness adjustment knob (3). The numerals by the lamp voltage indicator LEDs (4) indicate the voltage. Deselect the “PRESET” button (not illuminated green) to avoid using maximum illumination.
6. Adjust the interpupillar distance. While looking through the eyepieces, adjust the oculars (5) until the left and right fields of view coincide completely. 7. Put the 10x objective (6) in place. Place your specimen on the stage (7). 8. Find your specimen using the stage controls (9).
9. Focus specimen using fine/course focusing knobs (10). 10. Adjust the diopter:
- Close your left eye and focus on the specimen using the fine focus knob.
- Close your right eye and focus on the specimen using the diopter ring (11) on the left ocular.
- Open both eyes and confirm that the focus is comfortable.
11. If desired switch to the next objective by rotating the nosepiece (12) and focus. Continue until you reach the desired magnification. 12. Establish Koehler illumination:
- Close the field iris diaphragm (13) until you can see the edges.
- Focus the image of the field iris diaphragm by raising or lowering the condenser using the condenser height adjustment knob (14).
- Check if the circle of light is centered in the field of view. If not, use the two
condenser centering screws (15) to move the field iris diaphragm image to the center of the field of view.
- Open the field iris diaphragm until its image circumscribes the field of view.
- Match the opening of the condenser aperture iris diaphragm (16) with the N.A. of the objective in use in order to achieve the optimum objective performance. 13. Examine specimen and document image if necessary:
- To obtain still digitized images and movies, use the checklist on page _____ for the Sensicam Cooke Camera and ImagePro Plus software. 14. When finished:
- Lower the stage by turning the focus knob (10) and remove your specimen. - Turn the nosepiece (12) until the 10x objective is into place.
- Lower the light intensity to zero using the brightness adjustment knob (3). - Turn the halogen light switch (1) to “○” (OFF).
- Return any slide filters (26)(31)(32) to their disengaged positions.
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BRIGHTFIELD (reflected light)
Illumination is shone onto a reflective sample from above (the reflected light housing at the back of the microscope).
1. Turn on the halogen light switch (1) for reflected light to “│” (ON). Engage the brightfield mirror unit (BF) (17). 2. Adjust the light path.
- The light path selector knob (2) should be pushed in all the way. - The knob on the reflected light splitter (30) should be pushed in all the way.
- The shutter knob (20) should be slid to the position marked “○” (OPEN). 3. Select reflected light (upper position) on the switch by the brightness indicator (4). 4. Adjust the light intensity using the brightness adjustment knob (3). The numerals by the lamp voltage indicator LEDs (4) indicate the voltage. Deselect the“PRESET” button (not illuminated green) to avoid using maximum illumination.
5. Adjust the interpupillar distance. While looking through the eyepieces, adjust oculars (5) until the left and right fields of view coincide completely. 6. Put the 10x objective (6) in place. 7. Place your specimen on the stage (7).
8. Find your specimen using the stage controls (9).
9. Focus specimen using fine/course focusing knobs (10). 10. Adjust the diopter:
- Close your left eye and focus on the specimen using the fine focus knob.
- Close your right eye and focus on the specimen using the diopter ring (11) on the left ocular.
- Open both eyes and confirm that the focus is comfortable.
11. If desired switch to the next objective by rotating the nosepiece (12) and focus. Continue until you reach the desired magnification. 12. Establish Koehler illumination:
- Close the field iris diaphragm (13) until you can see the edges.
- Focus the image of the field iris diaphragm by raising or lowering the condenser using the condenser height adjustment knob (14).
- Check if the circle of light is centered in the field of view. If not, use the two
condenser centering screws (15) to move the field iris diaphragm image to the center of the field of view.
- Open the field iris diaphragm until its image circumscribes the field of view.
- Match the opening of the condenser aperture iris diaphragm (16) with the N.A. of the objective in use in order to achieve the optimum objective performance. 13. Examine specimen and document image if necessary:
- To obtain still digitized images and movies, use the checklist on page _____. 14. When finished:
- Lower the stage by turning the focus knob (10) and remove your specimen. - Turn the nosepiece (12) until the 10x objective is into place.
- Lower the light intensity to zero using the brightness adjustment knob (3). - Turn the halogen light switch (1) to “○” (OFF).
- Return any slide filters (26)(31)(32) to their disengaged positions.
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FLUORESCENCE
NOTE: The mercury bulb is fragile. It needs to be cool for at least 30 minutes before being turned on. And it needs to stay on for at least 15 minutes before being turned off. So check the usage log before turning it on and off. If you turn it on and off multiple times during the same session, please follow the guidelines.
1. Switch on the mercury bulb by setting the switch (18) on the Power Supply Unit to “I” (ON). The indicator (19) should light up. The mercury bulb will stabilize in 5-10 min. 2. Setup microscope for brightfield imaging (see instructions on ‘Brightfied reflected light’ page 2, steps 2-12). Use the reflected light splitter knob (30) to switch between the white and UV lamps that are located at the rear lamp housings.
3. Engage the fluorescence mirror unit (17) until the proper filter cube is in place: 1= Brightfield, 4 = TRITC, 5 = DAPI, 6 = FITC
4. When using the fluorescence mirrors, turn the light intensity down all the way using the brightness adjustment knob (3).
5. To allow light from the mercury bulb to reach the specimen, slide the shutter knob (20) to position marked “○” (OPEN). Close it whenever you wish to avoid photobleaching your sample.
6. To improve image contrast and to prevent color fading of fluorescent light in other part than observed region, pull out the field iris diaphragm knob (21) so that the image of the field iris diaphragm just circumscribes the field of view.
7. To help adjusting the brightness of the observed image and to improve the contrast, pull out the aperture iris diaphragm knob (22), the diaphragm will get smaller. 8. Examine specimen and document image if necessary:
- To obtain still digitized images and movies, use the checklist on page _____ for the Sensicam Cooke Camera and ImagePro Plus software. 9. When finished:
- Slide the shutter knob (20) to “●” (CLOSED).
- Push the knob on the reflected light splitter (30) in all the way.
- If mercury bulb has burned for at least 15 min., turn it off by setting the main switch (18) on the Power Supply Unit to “O”. The light (19) should go out. - Lower the stage by turning the focus knob (10) and remove the specimen. - Turn the nosepiece (12) back until the 10x objective is into place.
- Push the field iris diaphragm knob (21) and the aperture iris diaphragm knob (22) all the way back into the microscope.
- Engage the fluorescence mirror unit (17) back to BF (brightfield). - Return any slide filters (26)(31)(32) to their disengaged positions.
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(See diagram in printed operating manual)
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