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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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LED Thermo Flow — Combining Optogenetics with Flow Cytometry
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Two-photon in vivo flow cytometry using a fiber probe.

Yu-Chung Chang1, Jing Yong Ye2, Thommey P Thomas3

  • 1Center for Ultrafast Optical Science, University of Michigan, Ann Arbor, MI 48109.

Proceedings of Spie--The International Society for Optical Engineering
|May 17, 2016
PubMed
Summary

A novel double-clad fiber probe enables efficient two-photon excited flow cytometry in vitro and in vivo. This method overcomes blood scattering issues, achieving high detection efficiency for labeled cells in both buffer and whole blood.

Keywords:
Two-photon fluorescencecancercirculating tumor celldouble-clad fiberfiber probegreen fluorescent protein (GFP)in vivo flow cytometrymetastasis

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Area of Science:

  • Biomedical Optics
  • Flow Cytometry
  • In Vivo Imaging

Background:

  • Traditional flow cytometry faces challenges with scattering and absorption in biological samples like whole blood.
  • In vivo cellular analysis requires minimally invasive techniques for accurate detection.

Purpose of the Study:

  • To demonstrate a double-clad fiber probe for two-photon excited flow cytometry.
  • To evaluate the probe's performance in vitro and in vivo, particularly in whole blood.
  • To assess detection efficiency for fluorescently labeled cells in various conditions.

Main Methods:

  • Utilized a double-clad fiber probe for two-photon excitation.
  • Implemented two-channel fluorescence detection at distinct wavelengths.
  • Tested the system with DiD-labeled and green fluorescent protein (GFP)-expressing cells in PBS and whole blood.
  • Monitored cell circulation dynamics in live mice after injection.

Main Results:

  • Circumvented scattering and absorption issues inherent to whole blood.
  • Achieved consistent signal strength and detection efficiency for DiD-labeled cells in PBS and whole blood.
  • Demonstrated high detection efficiency for GFP-expressing cells in whole blood.
  • Observed in vivo detection efficiency of GFP-expressing cells consistent with in vitro whole blood results.

Conclusions:

  • The double-clad fiber probe is effective for two-photon excited flow cytometry in complex biological media.
  • This technique offers a robust solution for in vitro and in vivo cellular analysis, overcoming limitations of conventional methods.
  • The probe enables reliable monitoring of cell dynamics in live organisms.