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

Related Video (2)

Bilibili (China-Accessible Mirrors) ★ 72

Flow Cytometry Complete Workflow: Sample to Analysis

"End-to-end workflow including sample prep and troubleshooting, the context where tissue autofluorescence issues arise."

BD Biosciences ★ 70

Cell Preparation for Flow Cytometry

"Cell preparation best practices directly address reducing background and improving resolution for problematic samples."