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Possible errors
Weak fluorescence, insufficient brightness:
Wrongly stored, overaged or faded specimens.
Fast fading of the specimens (e. g. for FITC).
Unspecified filter combination.
Numerical aperture of the objective too low.
Eyepiece magnification too high.
Spent lamp.
Room too bright.
Trinocular tube: wrong beamsplitter setting.
Stray light due to reflections at the condenser.
Low-contrast image due to:
Excitation bandwidth too wide.
Inspecific staining.
Fluorescing mounting medium.
Autofluorescence of objective or immersion oil.
Glass surfaces dirty.