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Sidelobe suppressed Bessel beams for one-photon light-sheet microscopy.

Chetna Taneja1, Jerin Geogy George2, Stella Corsetti1

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Sidelobe suppressed Bessel beams (SSBB) enhance one-photon light-sheet microscopy by improving image contrast and reducing the need for deconvolution. This advanced Bessel beam (BB) technique offers superior imaging of biological samples.

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Area of Science:

  • Optics and Photonics
  • Biomedical Imaging
  • Microscopy

Background:

  • Bessel beams (BB) are widely used in light-sheet microscopy.
  • Standard BBs present challenges in one-photon fluorescence due to sidelobe effects.
  • Sidelobes in BBs can negatively impact image quality and resolution.

Purpose of the Study:

  • To develop and evaluate a sidelobe suppressed Bessel beam (SSBB) for enhanced one-photon light-sheet microscopy.
  • To compare the imaging performance of SSBB against standard BB.
  • To demonstrate the utility of SSBB in biological imaging applications.

Main Methods:

  • Utilized a computer-generated phase element to create an SSBB.
  • Performed comparative biological imaging experiments using SSBB and standard BB in a light-sheet configuration.
  • Assessed beam propagation depth and transverse size in phantom samples.
  • Imaged fixed early-stage zebrafish larvae to evaluate contrast-to-noise ratio (CNR).

Main Results:

  • The SSBB exhibited peak intensity over an order of magnitude higher than its first sidelobe.
  • SSBB light-sheets did not require deconvolution, unlike standard BB light-sheets.
  • SSBB maintained a 5 μm transverse size while propagating over 400 μm in phantom samples.
  • Imaging zebrafish larvae with SSBB resulted in a two-fold increase in CNR for cellular structures compared to standard BB.

Conclusions:

  • SSBB effectively mitigates sidelobe issues in one-photon light-sheet microscopy.
  • SSBB offers improved image contrast and clarity for biological samples.
  • This approach provides a valuable tool for enhancing contrast in one-photon light-sheet microscopy.