Excessive Autofluorescence in Solid Tissue Samples

Symptom

Solid tissue samples display much higher autofluorescence than PBMCs across multiple channels. Tissue-derived cells show elevated background due to structural proteins, extracellular matrix components, and pigments.

1. 1 Structural proteins and extracellular matrix components in solid tissues
2. 2 Tissue pigments contributing to broad-spectrum autofluorescence
3. 3 Epithelial cells containing structural proteins and pigments
4. 4 Tumor cells exhibiting elevated autofluorescence due to metabolic changes
5. 5 Aldehyde-based fixation further enhancing tissue autofluorescence

1. 1 Use spectral flow cytometry to model and subtract complex tissue autofluorescence signatures
2. 2 Choose dyes outside autofluorescence range (far-red, infrared) for key tissue markers
3. 3 Include unstained tissue controls to establish tissue-specific autofluorescence baseline
4. 4 Minimize fixation time and wash thoroughly to reduce aldehyde-enhanced autofluorescence
5. 5 Prioritize red/far-red channels (633 nm and beyond) for critical markers in tissue samples

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