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Updated: May 9, 2026
Flow Cytometry Analysis of Murine Bone Marrow Hematopoietic Stem and Progenitor Cells and Stromal Niche Cells
Published on: September 28, 2022
Frequency determination of rare populations by flow cytometry: a hematopoietic stem cell perspective
Alexandra Rundberg Nilsson1, David Bryder, Cornelis J H Pronk
1Immunology Unit, Institution for Experimental Medical Research, Lund University, 221 84, Lund, Sweden.
Insights
Multicolor flow cytometry enhances cellular subset identification, especially for rare cells like hematopoietic stem cells (HSCs). Strategies for sample analysis are crucial for accurate results in complex polychromatic flow cytometry.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Flow cytometry identifies cell subsets using fluorochrome-conjugated antibodies targeting cell-surface epitopes.
- Advances in instrumentation and fluorophores enable high-resolution multicolor analysis of cellular populations.
Purpose of the Study:
- To discuss considerations and strategies for multicolor flow cytometry.
- To exemplify analysis of human and mouse hematopoietic stem cells (HSCs).
Main Methods:
- Utilizing multicolor flow cytometry with fluorochrome-conjugated antibodies.
- Analyzing human and mouse hematopoietic stem cells (HSCs).
- Proposing strategies for sample analysis, including internal reference populations.
Main Results:
- Multicolor flow cytometry offers unprecedented resolution for identifying cellular populations.
- Analysis of rare subsets like HSCs presents significant challenges in sample preparation, acquisition, and analysis.
- Strategies such as using internal reference populations can aid in complex polychromatic flow cytometry.
Conclusions:
- Multicolor flow cytometry is a powerful tool for cellular subset identification.
- Careful consideration of preparation, acquisition, and analysis is vital for complex flow cytometry.
- Proposed strategies can improve the accuracy of identifying rare cellular subsets using polychromatic flow cytometry.
Abstract:
Flow cytometry allows for identification of cellular subsets based on cell intrinsic properties, most often by the use of fluorochrome-conjugated antibodies recognizing distinct cell-surface epitopes that define the cells of interest. Advances in technical instrumentation and the availability of an ever-increasing number of fluorophores, today enables identification of multicolor defined cellular populations to a previously unreachable resolution. However, these possibilities put an increasing demand on preparation, acquisition, and subsequent analysis of the investigated samples. Identification of very rare cellular subsets, such as the bone marrow-residing hematopoietic stem cells (HSCs), causes further complexity to such analysis. Here, we discuss considerations and aspects in multicolor flow cytometry as exemplified by analysis of human and mouse HSCs. We illustrate advantages and drawbacks of polychromatic flow cytometry and propose strategies, such as the use of internal reference populations, for sample analysis.
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