Chapter 3 Specific Operations
21
11.2
Selecting a Filter Cube
The epi fluorescence attachment can be equipped with the following filter cubes:
P-EFL GFP-B Filter Cube and P-EFL GFP-L Filter Cube: Used for fluorescence observation at a given wavelength.
P-EFLC Filter Cube: Used for fluorescence observation at an arbitrary wavelength when combined with arbitrary filters, for
example.
P-EFLBF Filter Cube (Bright Field): Used for episcopic bright-field observation.
Empty cubes (three cubes supplied with an epi-fluorescence attachment): Used when performing diascopic illumination
observation using the epi-fluorescence attachment.
11.3
Important Notes for Epi-Fluorescence Microscopy
Using non-fluorescent slide glass and cover glass
When performing fluorescence observation of samples with a slide glass or cover glass, be sure to use non-fluorescent
slide glass or cover glass to obtain a high-contrast image.
Locating a target in the sample
For epi-fluorescence microscopy, Nikon recommends locating a target using bright-field diascopic illumination (OCC
illumination) and then switching to epi-fluorescence, rather than emitting strong excitation light to the sample from the
beginning. When doing so, follow the operational tips below:
Start with low magnification under bright-field diascopic illumination.
If the sample is not transparent, use the P-EFLBF Filter Cube (Bright Field) to obtain the entire image by reflected light.
Gradually increase the magnification. When the target becomes difficult to locate, switch to epi-fluorescence and use
low excitation light.
Protection from ultraviolet light (light shielding plate)
The light shielding plate is used to prevent ultraviolet light
reflected by the sample, which is originally emitted through the
objective, from entering the observer’s eyes.
2 x
Apo
Plan
IN
MADE
JAPAN
Using the light shielding plate
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