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Updated: Jan 23, 2026

Measuring Diurnal Rhythms in Autophagic and Proteasomal Flux
Published on: September 17, 2019
High-throughput quantitative detection of basal autophagy and autophagic flux using image cytometry
Gopika SenthilKumar1, Justin H Skiba1, Randall J Kimple1
1Department of Human Oncology & UW Carbone Cancer Center, University of Wisconsin School of Medicine & Public Health, 750 Highland Ave, Madison, WI 53726, USA.
Abstract:
Quantitative assessment of changes in macro-autophagy is often performed through manual quantification of the number of LC3B foci in immunofluorescence microscopy images. This method is highly laborious, subject to image-field selection and foci-counting bias, and is not sensitive for analyzing changes in basal autophagy. Alternative methods such as flow cytometry and transmission electron microscopy require highly specialized, expensive instruments and time-consuming sample preparation. Immunoblots are prone to exposure-related variations and noise that prevents accurate quantification. We report a high-throughput, inexpensive, reliable and objective method for studying basal level and flux changes in late-stage autophagy using image cytometry and acridine orange staining.
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