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)

Leica Microsystems ★ 70

Laser Confocal Microscope | How To Set Up Confocal Multicolor Experiments

"Demonstrates setting up multicolor confocal acquisition, directly relevant to understanding and managing overlapping spectral contributions like broad lipofuscin autofluorescence."

Current Protocols ★ 62

Heterogeneous Autofluorescence

"Explains heterogeneous autofluorescence and spectral panel optimization, providing context for how age-related pigment signals complicate channel assignment."