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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

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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Imaging Flow Cytometry to Study Microbial Autoaggregation
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Flow cytometry without alignment of collection optics.

Greg Sitton1, Friedrich Srienc

  • 1Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, Minnesota 55455-0312, USA.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|October 6, 2009
PubMed
Summary

A novel Teflon AF (TFC) waveguide flow cell uses liquid core optics for cell analysis. This system enables efficient light collection and wavelength-specific measurements for applications like DNA content analysis.

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Area of Science:

  • Biophotonics
  • Optical Engineering
  • Analytical Chemistry

Background:

  • Traditional flow cytometry systems often rely on complex optical components like dichroic mirrors and collection objectives.
  • Developing cost-effective and efficient optical systems for cell analysis is crucial for advancing biological research and diagnostics.

Purpose of the Study:

  • To evaluate the performance of a new waveguide flow cell (TFC) for cell analysis.
  • To explore the use of fiber optic splitters as alternatives to conventional optical components.
  • To demonstrate the system's capability in measuring cellular properties like DNA content.

Main Methods:

  • Fabrication of a flow cell using Teflon AF (TFC), leveraging its low refractive index to create a liquid core waveguide.
  • Illumination of cells flowing through the TFC with a laser perpendicular to the flow direction.
  • Collection of scattered and fluorescent light via the TFC's axial path into a fiber optic.
  • Utilizing fiber optic splitters to divide the collected light for multi-wavelength analysis.
  • Employing optical filters and photomultiplier tubes (PMTs) for specific wavelength detection.
  • Testing the system with polystyrene beads for alignment and performance assessment, and with CHO and yeast cells for DNA content measurement.

Main Results:

  • The TFC demonstrated effective light collection due to its liquid core waveguide properties.
  • Polystyrene beads exhibited unique bimodal light scattering signals dependent on flow and collection geometry, aiding system characterization.
  • The system successfully measured DNA content in CHO and yeast cells, validating its analytical capabilities.
  • Fiber optic splitters proved viable for spectral separation, potentially simplifying optical setups.

Conclusions:

  • The TFC-based waveguide flow cell offers a promising, potentially lower-cost alternative for cell analysis.
  • Fiber optic splitters can effectively replace traditional collection optics and dichroic mirrors in certain flow cytometry applications.
  • The system's ability to perform spectral measurements highlights its versatility for various biological assays.