Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry.

摘要: Autophagy is a catabolic pathway in which normal or dysfunctional cellular components that accumulate during growth and differentiation are degraded via the lysosome recycled. During autophagy, cytoplasmic LC3 protein lipidated recruited to autophagosomal membranes. The autophagosome then fuses with form autolysosome, where breakdown of vesicle its contents occurs. ubiquitin-associated p62, binds LC3, also used monitor autophagic flux. Cells undergoing autophagy should demonstrate co-localization lysosomal markers. Immunofluorescence microscopy has been visually identify puncta, and/or lysosomes on per-cell basis. However, an objective statistically rigorous assessment can be difficult obtain. To overcome these problems, multispectral imaging flow cytometry was along analytical feature compares bright detail images from three markers (LC3, p62 LAMP1) quantifies their co-localization, combination spot counting measure objective, quantitative, robust manner.