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

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.