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Non-linear optical flow cytometry using a scanned, Bessel beam light-sheet.

Bradley B Collier1, Samir Awasthi1,2, Deborah K Lieu3

  • 1Center for Biophotonics, University of California, Davis.

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|May 30, 2015
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Summary

Label-free cell analysis using non-linear optical (NLO) phenomena is gaining interest. A scanned Bessel beam light-sheet optimizes NLO signal generation in microfluidics, outperforming conventional methods for enhanced cellular analysis.

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Area of Science:

  • Biophysics
  • Optical Engineering
  • Cellular Biology

Background:

  • Flow cytometry is crucial for high-throughput single-cell analysis.
  • Current methods often rely on cellular labeling, which presents challenges.
  • Label-free cellular analysis is increasingly desired for intrinsic biomolecular measurement.

Purpose of the Study:

  • To investigate non-linear optical (NLO) phenomena for label-free cellular analysis.
  • To determine the optimal excitation geometry for efficient NLO signal generation in microfluidics.
  • To compare NLO signal generation using different excitation focal geometries.

Main Methods:

  • Utilized a microfluidic environment for cellular analysis.
  • Employed a light-sheet excitation geometry consisting of a scanned Bessel beam.
  • Measured two-photon fluorescence intensities in a model polystyrene microparticle system.
  • Compared results with a conventional Gaussian excitation beam.

Main Results:

  • A scanned Bessel beam light-sheet proved optimal for NLO signal generation.
  • This geometry provided an effective balance of photon density and cross-sectional area.
  • Significantly larger two-photon fluorescence intensities were measured compared to Gaussian excitation.
  • Demonstrated the potential for label-free intrinsic optical measurements.

Conclusions:

  • A Bessel beam light-sheet is a highly efficient excitation geometry for NLO-based label-free cellular analysis in microfluidics.
  • This approach offers advantages over conventional flow cytometry excitation methods.
  • The findings support the development of advanced label-free cellular analysis techniques.