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Chapter 3 Operation Details
51
3.19.2 Excitation
light
filter (EX filter)
An excitation light filter transmits lights selectively and
blocks other lights. The transmitted lights are called
excitation lights. They are used to excite the
fluorophore in the sample and fluorescent lights are
emitted from the sample. The wavelength range of
lights that can pass through the filter is called the
bandwidth.
The bandwidth of the excitation light filter determines
the brightness of the fluorescent image, the
occurrence of autofluorescence (fluorescence
resulting from substances other than the
fluorophores), and degree of fading. When the filter
has a wide bandwidth, a large amount of excitation
lights will be irradiated on the sample. In this case,
the image becomes bright but the amount of
autofluorescence becomes large and fading of the
sample occurs soon. On the contrary, when the filter
has a narrow bandwidth, a small amount of excitation
lights will be irradiated on the sample. In this case,
the image becomes dark but the amount of
autofluorescence becomes small and fading of the
sample occurs late. For samples with pronounced
autofluorescence, use an excitation light filter with a
narrow bandwidth. (The resulting fluorescent image
will be darker, however.)
The excitation light filter is exposed to strong lights.
Therefore it may deteriorate under use. Please
replace it at a proper interval based on the hours
used.
Narrow
Bandwidth of excitation filter
Wide
Brightness of
fluorescence image
Dark Bright
Occurrence of
autofluorescence
Less frequent
Frequent
Degree of fading
Small
Large
EX filter
Bandwidth
Wavelength
Spectral
transmittance
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