True. Autofluorescence is a significant challenge in immunofluorescence (IF) microscopy. It refers to the natural emission of light by certain endogenous tissue components when excited by the wavelengths used for fluorescence imaging. Common sources include: 1) **Red blood cells** (due to porphyrins), which can emit a dull red fluorescence. 2) **Elastic fibers** in blood vessels and skin, which often autofluoresce a bright greenish-yellow. 3) **Lipofuscin** (a 'wear-and-tear' pigment in aging cells), which has broad emission. 4) **Collagen** and some fixatives (like glutaraldehyde). This inherent glow can be mistaken for specific antibody staining, leading to false-positive interpretations. To mitigate this, pathologists use several strategies: using dyes that quench autofluorescence (like Sudan Black B or TrueBlack), choosing fluorophores with emission spectra distinct from the autofluorescence, using spectral imaging to unmix signals, and always comparing the IF stain to a negative control slide processed without primary antibody to identify background autofluorescence patterns.

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