An optimised protocol for the detection of lipofuscin, a versatile and quantifiable marker of cellular senescence.

ABSTRACT: Lipofuscin is a yellow-brown pigment typically found in the lysosomes that contains a mixture of molecules including lipids, metals and misfolded proteins. The use of Sudan black B to detect lipofuscin accumulation, a well described marker of cellular senescence and ageing, was first described in 2013 by Georgakopoulou, et al. Here, we provide an optimisation of the original protocol. Firstly, we adjusted the staining methodology for increased ease of use on cultured cells. Secondly, we show that Sudan black B-stained lipofuscin emits strong fluorescence in the far-red channel making it suitable for fluorescence microscopy detection and quantification. Moreover, we also demonstrate that this optimised protocol can be utilised in conjunction with standard immunofluorescence staining techniq

SUBMITTER: Davan-Wetton CSA 

PROVIDER: S-EPMC11249248 | biostudies-literature | 2024

REPOSITORIES: biostudies-literature