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Chapter 3
Individual Operations
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Tips on Microscope Settings for Photomicroscopy
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Adjusting the light intensity
Lamp voltage: When accurate color reproduction is critical, turn the brightness control knob to the
mark, bring
the NCB11 filter into the optical path, and use the ND filters to adjust the brightness.
Filter:
Place a commercially available color compensation filter on the field lens at the microscope base, as
necessary.
■
Adjusting
the
condenser
Always focus and center the condenser.
For differential interference contrast microscopy, align the orientation of the polarizer and analyzer.
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 sample, making it more difficult to acquire
suitable fluorescent images. Insert ND filters into the optical path to adjust the brightness.
Specimen
Photomicrography of faded specimen sections requires prolonged exposure time and results in poor color
reproduction and low-quality images. Move the specimen to obtain images from a fresh section of the specimen
previously unexposed to excitation light. For best results, use the phase contrast method to select a specimen
section for photomicrography, and then switch to the fluorescent method to capture images.
■
Adjusting the brightness of the image on the monitor
When observing images captured by the camera and displayed on the monitor, you can adjust the brightness by
varying camera adjustment parameters, such as display mode, exposure mode, photometry mode, exposure
compensation, and image level adjustment.
See the instruction manual for the DS-U3, DS-L3, or the microscope's setup/control software “Ni Setup Tool” for
details.
Содержание ECLIPSE Ni-E
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