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Passive optical separation within a 'nondiffracting' light beam.

Lynn Paterson1, Eirini Papagiakoumou, Graham Milne

  • 1University of Saint Andrews, Bute Medical School, Bute Building, Saint Andrews, Fife KY16 9TS, United Kingdom. L.Paterson@hw.ac.uk

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|November 13, 2007
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Summary
This summary is machine-generated.

This study introduces a passive optical cell sorter using Bessel beams to separate cells based on size and refractive index. Cell viability is confirmed post-sorting, demonstrating potential for biological applications.

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Area of Science:

  • Biophysics
  • Optical Engineering
  • Cell Biology

Background:

  • Passive optical cell sorting offers a label-free method for cell separation.
  • Bessel beams, as nondiffracting light patterns, provide a stable optical field for manipulation.
  • Particle properties like size and refractive index influence interaction with light fields.

Purpose of the Study:

  • To develop and evaluate a passive optical cell sorter utilizing Bessel beams.
  • To assess the sorter's resolution based on particle size and refractive index.
  • To demonstrate the separation of cells and confirm their viability post-sorting.

Main Methods:

  • A passive optical cell sorter was constructed using a Bessel beam.
  • Microspheres of varying sizes and refractive indices were used to calibrate the sorter.
  • Native cell populations (erythrocytes, lymphocytes, HL60, murine bone marrow, stem/progenitor cells) were tested.
  • Silica microsphere tags were applied to cells to enhance optical properties for sorting.

Main Results:

  • Microsphere migration time in the Bessel beam correlated with size and refractive index.
  • Successful separation of erythrocytes from lymphocytes was achieved based on intrinsic size differences.
  • Passive separation of other tested cell types was insufficient due to small intrinsic differences.
  • Labeled cells, modified with silica microspheres, were successfully separated.
  • Sorted cells demonstrated high viability, confirmed by trypan blue exclusion, in vitro cloning, and lack of apoptosis markers.

Conclusions:

  • Passive optical sorting with Bessel beams can separate cells based on optical properties.
  • Particle tagging is a viable strategy to enhance separation of cells with similar intrinsic properties.
  • The developed cell sorter preserves cell viability, indicating its potential for downstream biological applications.