Nikon Eclipse Ni-E Instruction Manual page 100

Tips on Microscope Settings for Photomicroscopy
■ Adjusting the light intensity
Lamp voltage: When accurate color reproduction is critical, turn the brightness control knob to the
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.
• 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 samples 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.
• Sample
Photomicrography of faded sample sections requires prolonged exposure time and results in poor color
reproduction and low-quality images. Move the sample to obtain images from a fresh section of the sample
previously unexposed to excitation light. For best results, use the differential interference contrast microscopy
method to select a sample 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,
compensation, and image level adjustment.
See the instruction manual provided with the DS-U3, DS-L3, or the microscope's setup/control software "Ni Setup Tool"
for details.
Chapter 3
Individual Operations
such as display mode, exposure mode, photometry mode, exposure
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