Biopharma Insights Here, multispectral imaging flow cytometry with an analytical feature that compares bright detail images of 3 autophagy markers and quantifies their co-localization, along with LC3 spot counting, was used to measure autophagy in an objective, quantitative, and statistically robust manner.
Total time
~2 days (labeling, acquisition, and analysis per sample set)
Model organism
Jurkat cells (human T lymphocyte cell line)
Steps
1
Label Jurkat cells for autophagy markers
Stain Jurkat cells with fluorescent antibodies or probes against LC3, p62, and LAMP1 to enable multiplexed detection.
▶ 00:42
2
Acquire cells on multispectral imaging flow cytometer
Run labeled samples on the MIFC instrument to capture bright detail images of each autophagy marker.
▶ 02:18
3
Analyze co-localization and LC3 spots
Use the analytical feature to compare bright detail images, quantify marker co-localization, and count LC3 spots.
▶ 03:43
4
Interpret representative MIFC results
Review representative analyses demonstrating objective, quantitative measurement of autophagic flux.
▶ 08:35