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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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Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction
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Label-free multiphoton imaging flow cytometry.

Ryo Kinegawa1, Julia Gala de Pablo1, Yi Wang2

  • 1Department of Chemistry, The University of Tokyo, Tokyo, Japan.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|February 17, 2023
PubMed
Summary
This summary is machine-generated.

A new multiphoton imaging flow cytometry visualizes starch and collagen in live cells without labels. This high-throughput method advances biological studies and cell strain optimization.

Keywords:
Muriellaimaging flow cytometrymicroalgaenonlinear opticssecond harmonic generation

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Microscopy

Background:

  • Conventional fluorescence-based imaging flow cytometry relies on fluorescent labeling, posing technical challenges.
  • Existing label-free methods like optofluidic time-stretch quantitative phase imaging and stimulated Raman scattering (SRS) imaging cannot probe molecules such as starch and collagen.

Purpose of the Study:

  • To introduce a novel label-free imaging flow cytometry technique, multiphoton imaging flow cytometry, for visualizing starch and collagen in live cells.
  • To demonstrate the high-throughput capability of this new method for biological research.

Main Methods:

  • Developed a multiphoton imaging flow cytometer utilizing nonlinear optical imaging with four-wave mixing (FWM) and second-harmonic generation (SHG) for contrast.
  • Integrated a microfluidic chip with an acoustic focuser, a laser scanning SHG-FWM microscope, and high-speed image acquisition.
  • Achieved image acquisition of 100x100 pixels with 500nm spatial resolution and a 50μm x 50μm field of view.

Main Results:

  • The system achieves a high event rate of 4-5 events/second, corresponding to a throughput of 560-700 kb/s.
  • Successfully visualized starch and collagen in live cells, including starch distribution dynamics in Chromochloris zofingiensis.
  • Demonstrated the utility for statistical high-content analysis of biological functions.

Conclusions:

  • Multiphoton imaging flow cytometry offers a powerful label-free approach for visualizing key biomolecules like starch and collagen.
  • This technique enables high-throughput, high-content analysis crucial for understanding cellular functions and optimizing cell strains for industrial applications.