effective in finding your sample without subjecting it to fluorescence. Focus of the condenser may need
to be re-adjusted if changing the level of focus in the sample or after switching objectives.
1. Place a sample on the stage and obtain an image using brightfield or DIC to the best possible focus.
2. Make sure the Condenser Aperture Diaphragm, Figure 13.1 is fully open by moving the black lever
fully to the left;
3. Observe through the eyepieces as the Field Stop Diaphragm, Figure 13.3, is closed down by moving
the black lever to the right;
4. If closing the field stop blocks the light, stop and use the 2 centering screws, Figure 13.5, to move
the aperture image back into view.
5. Once the Field Stop is fully closed, rotate the condenser focus knob, Figure 13.2, until the edges of
the iris are as sharp as possible;
6. Use the centering screws to move the field stop into the center of the field of view;
7. Open the field stop just until the edges almost disappear from the field of view;
8. Now, close down the condenser aperture diaphragm just until you begin to see the image dim
slightly, then nudge it back open until the light intensity is restored;
9. Check the focus of the sample, it may change if the condenser was initially out of focus;
10. If the focus was changed, re-check the condenser by simply closing the field stop and finely
adjusting the condenser focus, then re-open the field stop;
11. Now, the condenser is properly adjusted, for this objective lens, to provide sharp transmitted light
images or to direct unabsorbed laser light to the transmitted light detector.
12. If collecting an image with the transmitted light detector for confocal imaging, focus on some level
within the specimen by confocal. Then switch to transmitted light imaging through the eyepieces to
set Koehler illumination.
7.3.1 Condenser position.
The condenser is moved out of the way for inserting a specimen by either turning the condenser
focus knob or by tilting. It tilts on a hinge located behind it, at the support. It must be in the
horizontal position for any procedure requiring transmitted light. Keep it tilted down in working
position when performing confocal microscopy to avoid the possibility of vibration should it slip
down on its own.
Figure 14. Tilting the condenser.
Olympus Fluoview-1000 User’s Guide
V.M. Bloedel Hearing Research Center, Core for Communication Research
Center on Human Development and Disability, Digital Microscopy Center
May 11, 2011 43
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