Chapter 1 Microscopy Procedures
30
Chapter 1-5
Microscopy Procedures
Epi-fluorescence Microscopy
7
Bring an arbitrary objective into the optical path.
Turn the nosepiece to bring the desired objective into the optical path.
When using an oil immersion type objective, apply immersion oil between the specimen and the objective.
(→Chapter 2 “12 Oil Immersion” for details)
Non-fluorescent immersion oil
Use Nikon designated non-fluorescent immersion oil.
8
Focus on the specimen.
(1) Look into the eyepiece, and adjust the brightness o
the field of view with the ND filter of the
epi-fluorescence attachment.
(2) Rotate the stage knob to move the stage and bring
the target into the optical path.
(3) If the specimen is not in focus, turn the focus knob to
focus on it.
ND4
ND8
ND16
A
1
-
2
-
3
-
4
B
1
-
2 / 3
-
4
C
1
-
2
-
3
-
4
Adjusting the brightness with ND filters
9
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 almost
circumscribes the field of view.
Size of the field diaphragm
Normally, adjust the field diaphragm so that it
almost circumscribes the field of view. Opening
the field diaphragm too much results in stray light
entering the field of view, generating flare and
reducing the image contrast. In addition, the
specimen will become decolorized over a wide
area.
Field diaphragm's adjustment timing
Be sure to adjust the field diaphragm each time
you change the objective.
A1
-
2
-
3
-
4
B
1
-
2 / 3
-
4
C1
-
2
-
3
-
4
1
1
4
2
3
4
CUBE
Circumscribe around the field of view
Adjusting the field diaphragm
Field diaphragm
lever
Field of view
Field diaphragm
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