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Flow Cytometry01:23

Flow Cytometry

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The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Related Experiment Video

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Simultaneous Assessment of Kinship, Division Number, and Phenotype via Flow Cytometry for Hematopoietic Stem and Progenitor Cells
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Development of a Human Whole Blood Screening Platform to Monitor JAK/STAT Signaling Using High-Throughput Flow

Mark P Fereshteh1, Xin Li2, Sha Li2

  • 1Bristol-Myers Squibb Company, Princeton, NJ, USA Mark.Fereshteh@bms.com.

Journal of Biomolecular Screening
|May 5, 2016
PubMed
Summary

Researchers developed a high-throughput assay to screen Janus-associated kinase (JAK) inhibitors in human blood. This new method increases efficiency and enables the discovery of novel JAK/STAT signaling inhibitors.

Keywords:
JAK/STAT signalingautomationflow cytometryhigh throughputhuman whole bloodminiaturization

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Area of Science:

  • Pharmacology and Drug Discovery
  • Immunology
  • Biotechnology

Background:

  • Oral Janus-associated kinase (JAK) inhibitors are under clinical development for various conditions.
  • Assessing JAK inhibition and its biological effects in human blood is crucial but traditionally low-throughput and expensive.
  • Existing methods for evaluating JAK/STAT signaling in blood lack efficiency for large-scale screening.

Purpose of the Study:

  • To develop a high-throughput, flow-based assay for screening small molecules targeting JAK/STAT signaling in human whole blood.
  • To enable sensitive measurement of target engagement and biological outcomes of JAK inhibitors.
  • To overcome limitations of conventional manual flow cytometry for analyzing signaling events in blood.

Main Methods:

  • Development of a robust 384-well, high-throughput flow-based assay platform.
  • Utilized automation technologies and process optimizations to address sample handling and data analysis challenges.
  • Evaluated small molecule compounds targeting JAKs in peripheral human whole blood.

Main Results:

  • The developed assay provides a highly sensitive analysis of JAK/STAT signaling events in human blood.
  • Automation and optimized processes improved sample throughput significantly compared to manual methods.
  • The assay platform successfully facilitated the measurement of target engagement for JAK inhibitors.

Conclusions:

  • A novel, high-throughput flow-based assay for screening JAK/STAT inhibitors in human whole blood has been established.
  • This platform overcomes previous throughput and cost limitations, enabling efficient drug discovery.
  • The assay facilitates the development and identification of novel JAK/STAT pathway inhibitors.