Chapter 2 Microscopy Operations
39
Epi-fluorescence
Microscopy
Preparation Focusing Microscopy
Epi-fluorescence
Microscopy Procedure
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10
Adjust the field diaphragm of the epi-fluorescence attachment.
Use the field diaphragm lever of the epi-fluorescence
attachment to adjust the field diaphragm so that it
circumscribes the field of view.
Size of the field diaphragm
Normally, adjust the field diaphragm so that it
circumscribes the field of view. Excessively opening
the field diaphragm will result in stray light entering
the field of view, generating flare and reducing the
image contrast. In addition, the sample will become
decolorized over a wider area.
Field diaphragm's adjustment timing
Be sure to adjust the field diaphragm each time you
change the objective.
Appropriate size of the field diaphragm
Adjusting the field diaphragm
11
Adjust the brightness.
Use ND filters on the epi-fluorescence attachment to
adjust image brightness.
The aperture diaphragm of the epi-fluorescence
attachment can also be used for brightness adjustment
of the image. Be sure to center the aperture diaphragm
before use. (See Chapter 3 “16.4 Adjusting the
Brightness of the Fluorescent Image (Using ND Filters
and the Aperture Diaphragm)”.)
ND4
ND8
ND16
Brightness adjustment using the ND filter
Field diaphragm
Field of view
Circumscribe around the field of view.
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