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Immunophenotyping and Cell Sorting of Human MKs from Human Primary Sources or Differentiated In Vitro from Hematopoietic Progenitors
Published on: August 7, 2021
7.0K
High-speed fluorescence image-enabled cell sorting
Daniel Schraivogel1, Terra M Kuhn2, Benedikt Rauscher1
1Genome Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
Science (New York, N.Y.)
|January 20, 2022
Summary
High-speed image-enabled cell sorting (ICS) precisely isolates single cells based on morphology and protein localization. This advanced cell sorting technology accelerates genome-wide screening and expands phenotypic analysis capabilities.
Area of Science:
- Cell Biology
- Biotechnology
- Genomics
Background:
- Isolating single cells with specific spatial and morphological traits is a significant technical hurdle.
- Current cell sorting methods lack the resolution to capture complex cellular phenotypes.
Purpose of the Study:
- To develop a high-speed image-enabled cell sorting (ICS) method for precise single-cell isolation.
- To expand the capabilities of cell sorting and pooled genetic screening through advanced phenotypic analysis.
Main Methods:
- Established high-speed image-enabled cell sorting (ICS) capable of recording multicolor fluorescence images and sorting cells at 15,000 events/sec.
- Utilized ICS to quantify cell morphology, protein localization, and mitotic stages for enhanced cell cycle analysis.
- Combined ICS with CRISPR-pooled screens for rapid genome-wide screening.
Main Results:
- ICS successfully quantifies cell morphology and protein localization, improving resolution in cell cycle analyses.
- Enabled completion of genome-wide image-based screens in approximately 9 hours.
- Demonstrated identification of nuclear factor κB (NF-κB) pathway regulators.
Conclusions:
- ICS significantly broadens the phenotypic space accessible for cell sorting and pooled genetic screening.
- This technology offers a powerful tool for analyzing complex cellular phenotypes and accelerating biological discovery.
