Tissue autofluorescence may obscure the desired target during immunofluorescence. A number of interventions may reduce this interference.
Autofluorescence can be detected by examining untreated tissue using the immunofluorescent microscope.
Problem
Solution
use of aldehyde fixative (glutaraldehyde, other)
Avoid use of aldehyde fixative. Consider use of acetone-fixed frozen section.
tissue autofluorescence, other
Treat with a fluorescent background dye to mask the autofluorescence prior to application of primary immunoreagent.
Evaluate different filter set-ups to see if the nonspecific autofluorescence can be masked.
The differential diagnosis is nonspecific uptake of an immunoreagent.
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Specialty: Clinical Laboratory
