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Related Concept Videos

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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Color-scalable flow cytometry with Raman tags.

Ryo Nishiyama1, Kotaro Hiramatsu1,2,3, Shintaro Kawamura4,5

  • 1Department of Chemistry, The University of Tokyo, Tokyo 113-0033, Japan.

PNAS Nexus
|February 27, 2023
PubMed
Summary
This summary is machine-generated.

Researchers developed color-scalable flow cytometry using Raman tags to overcome the color barrier, enabling analysis of over 140 cell characteristics. This advanced technique enhances cell analysis in biology and medicine.

Keywords:
Raman probeRaman spectroscopyRaman tagendocytosisflow cytometry

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Cell Biology

Background:

  • Flow cytometry is crucial for cell analysis but limited by spectral overlap, restricting the number of simultaneously resolved cell characteristics.
  • The "color barrier" in traditional flow cytometry arises from overlapping fluorescence signals, limiting multiplexing capabilities.

Purpose of the Study:

  • To overcome the limitations of traditional flow cytometry by developing a color-scalable method.
  • To enable high-multiplexing cell analysis beyond the constraints of spectral overlap.

Main Methods:

  • Utilized coherent Raman flow cytometry with novel resonance-enhanced cyanine-based Raman tags and Raman-active dots (Rdots).
  • Synthesized 20 linearly independent cyanine-based Raman tags and Rdots containing 12 different Raman tags for sensitive detection.
  • Employed a broadband Fourier-transform coherent anti-Stokes Raman scattering (FT-CARS) flow cytometer.

Main Results:

  • Achieved a detection limit as low as 12 nM for Rdots with a short integration time (420 µs).
  • Demonstrated high-accuracy (98%) multiplex flow cytometry of MCF-7 breast cancer cells using 12 Rdots.
  • Successfully performed large-scale time-course analysis of endocytosis using the multiplex Raman flow cytometer.

Conclusions:

  • The developed color-scalable flow cytometry breaks the "color barrier" inherent in traditional methods.
  • This technique theoretically allows for multiplex flow cytometry of live cells with over 140 colors using a single laser and detector.
  • Offers a cost-effective and scalable solution for advanced cell analysis without increasing instrument complexity.