Light sheet fluorescence microscopy using axi-symmetric binary phase filters

Area of Science:

• Biomedical Optics
• Microscopy Techniques
• Optical Engineering

Background:

• Light sheet fluorescence microscopy (LSFM) offers depth-sectioning and low photobleaching for biological studies.
• Axial resolution in LSFM depends on illumination sheet thickness, necessitating high numerical aperture lenses.
• Illumination beam divergence limits the field-of-view (FoV), hindering high-resolution imaging over larger areas.

Purpose of the Study:

• To develop an enhanced LSFM technique for improved field-of-view (FoV) and imaging performance.
• To investigate the utility of binary phase filters (BPFs) derived from axi-symmetric aberrations for LSFM.
• To overcome limitations of complex optical setups for generating specialized beams like Bessel or Airy beams.

Main Methods:

• Integration of binary phase filters (BPFs) into the LSFM illumination path.
• Utilizing BPFs derived from phase axicon and spherical aberrations.
• Demonstrating imaging of microspheres, Drosophila larva imaginal discs, and Arabidopsis samples.

Main Results:

• The novel LSFM approach significantly increased the effective field-of-view (FoV).
• BPFs derived from axicon aberrations generated self-reconstructing beams, beneficial for imaging scattering specimens.
• High-contrast, three-dimensional imaging was achieved over an extended FoV.

Conclusions:

• Binary phase filters offer a practical and effective method for enhancing FoV in LSFM.
• The developed LSFM technique improves imaging capabilities for complex biological samples.
• This approach provides a valuable tool for advanced biomedical imaging research.