Multicolor flow cytometry-based quantification of mitochondria and lysosomes in T cells

Chin Wen Wei, Tyng An Zhou, Ivan L. Dzhagalov, Chia Lin Hsu*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Scopus citations


T cells utilize different metabolic programs to match their functional needs during differentiation and proliferation. Mitochondria are crucial cellular components responsible for supplying cell energy; however, excess mitochondria also produce reactive oxygen species (ROS) that could cause cell death. Therefore, the number of mitochondria must constantly be adjusted to fit the needs of the cells. This dynamic regulation is achieved in part through the function of lysosomes that remove surplus/damaged organelles and macromolecules. Hence, cellular mitochondrial and lysosomal contents are key indicators to evaluate the metabolic adjustment of cells. With the development of probes for organelles, wellcharacterized lysosome or mitochondria-specific dyes have become available in various formats to label cellular lysosomes and mitochondria. Multicolor flow cytometry is a common tool to profile cell phenotypes, and has the capability to be integrated with other assays. Here, we present a detailed protocol of how to combine organelle-specific dyes with surface markers staining to measure the amount of lysosomes and mitochondria in different T cell populations on a flow cytometer.

Original languageEnglish
Article numbere58844
JournalJournal of Visualized Experiments
Issue number143
StatePublished - Jan 2019


  • Flow cytometry
  • Immunology and infection
  • Issue 143
  • Lysosome
  • Mitochondria
  • Multicolor
  • Organelle-specific dyes
  • T cell


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