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(7)
Align the specimen.
(8)
Readjust the focus onto the target.
(9)
Adjust the image brightness using the camera exposure compensation function.
(10) Check the image using the [FREEZE] button.
(11) If the image is acceptable, press the Capture button to save the image.
(The operating procedure differs if the DF/FL scene mode is selected. For details, refer to the instruction manual
provided with the camera.)
Number of images captured
Seriography is not available because the Capture button is designed to capture only one image.
Tips on Microscope Settings for Photomicroscopy
18.2
Adjusting the light intensity
LED: Adjust the brightness using the brightness control knob.
Filter: Place a commercially available color compensation filter on the field lens on the microscope base, inside or on the
filter cassette holder as necessary.
Adjusting the condenser
Always focus and center the condenser.
For phase contrast microscopy, center the annular diaphragm.
The diaphragm aperture should generally be adjusted to 70 to 80% of the numerical aperture of the objective.
Confirming the photomicrographic range
The image on the monitor represents the photomicrographic range.
Confirming the focus
Check the focus both through the eyepiece and on the monitor. If the focal positions for the two images differ, adjust the
camera fine focus adjustment ring at the camera port.
Adjusting to keep out extraneous light
Field diaphragm: Stop down the diaphragm to a setting just slightly wider than the area shown on the monitor.
Eyepiece:
Cover the eyepiece with a piece of cloth or similar.
Protecting fluorescent images from decoloration
The fluorescence of specimens may fade during exposure. To prevent this, do the following:
■
Use a brighter optical system combination
Even if the overall magnification is the same on the monitor, the combination of objective and camera zoom can
result in significant variations in exposure time. Nikon recommends increasing the magnification of the objective
rather than of the zoom. (Generally, the numerical aperture of the objective increases with magnification. The larger
the numerical aperture, the brighter the resulting image.)
■
Adjusting the excitation light
Excessively bright excitation light will accelerate the decoloration of the specimen, making it more difficult to acquire
suitable fluorescent images. Insert ND filters into the optical path to adjust the brightness.
Chapter 2
Individual Operations
65
Chap. 2
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