Protocol to identify and analyze mouse and human quiescent hematopoietic stem cells using flow cytometry combined with confocal imaging

Jiajing Qiu, Vijay Menon, Nikolaos Tzavaras, Raymond Liang, Saghi Ghaffari

Research output: Contribution to journalArticlepeer-review

Abstract

Mitochondrial membrane potential (MMP) segregates functionally distinct subsets within highly purified hematopoietic stem cells (HSCs). Here, we detail a protocol for FACS isolation of MMP sub-fractions of phenotypically defined mouse and human HSCs. These steps are followed by high-/super-resolution immunofluorescence microscopy of HSCs’ lysosomes. While the protocol describes the isolation of quiescent HSCs, which are the most potent subsets, it could also be applied to other HSC subsets. This protocol overcomes some experimental challenges associated with low HSC numbers. For complete details on the use and execution of this protocol, please refer to Liang et al. (2020) and Qiu et al. (2021).

Original languageEnglish
Article number101828
JournalSTAR Protocols
Volume3
Issue number4
DOIs
StatePublished - 16 Dec 2022

Keywords

  • Cell Biology
  • Cell culture
  • Cell isolation
  • Flow Cytometry/Mass Cytometry
  • Metabolism
  • Microscopy
  • Stem Cells

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