Age-Related Lipofuscin Accumulation

Symptom

Exceptionally broad excitation and emission spectra affecting multiple channels in aged tissues such as brain, heart, skeletal muscle, and retina. Age-dependent lysosomal pigment cannot be confined to single channel.

1. 1 Age-dependent accumulation of lipofuscin in lysosomes
2. 2 Broad, overlapping excitation and emission spectra
3. 3 Particularly prominent in metabolically active tissues
4. 4 Increased accumulation in aged specimens

1. 1 Apply Sudan Black B (0.05-0.3% w/v in 70% EtOH for ~10 min) as primary quencher
2. 2 Rinse thoroughly and mount in aqueous medium (SBB is alcohol-soluble)
3. 3 Assign dim targets exclusively to far-red channels
4. 4 Document autofluorescence baseline with no-primary controls in each channel
5. 5 Consider spectral unmixing for mathematical separation of autofluorescence

Related Video (2)

Current Protocols ★ 92

Deciding on an Approach for Mitigating Autofluorescence

"Directly addresses autofluorescence mitigation strategies, the core problem in this failure case of lipofuscin-induced spectral spillover"

JoVE (Open Access) ★ 78

Measuring Interactions between Fluorescent Probes and Lignin in Plant Sections by sFLIM Based on Native Autofluorescence

"Demonstrates sFLIM technique for handling native autofluorescence in tissue imaging, directly applicable to age-related lipofuscin interference across multiple channels"