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Benchtop mesoSPIM: a next-generation open-source light-sheet microscope for cleared samples.

Nikita Vladimirov1,2,3, Fabian F Voigt4,5,6, Thomas Naert7

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The next-generation mesoSPIM microscope offers enhanced capabilities for imaging large cleared tissues. This open-source light-sheet microscopy system provides higher resolution, faster imaging, and supports larger samples for diverse scientific applications.

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Area of Science:

  • Biophysics
  • Optical Engineering
  • Biological Imaging

Background:

  • The original mesoSPIM initiative (2015) aimed to democratize light-sheet microscopy for large cleared tissues.
  • Increasing demands for high-speed, high-resolution imaging of larger biological samples necessitate advanced microscopy solutions.

Purpose of the Study:

  • Introduce the next-generation mesoSPIM (Benchtop) microscope.
  • Significantly enhance imaging capabilities for large-scale biological samples.
  • Improve accessibility, affordability, and ease of assembly for light-sheet microscopy.

Main Methods:

  • Developed a novel optical method for evaluating detection objectives tailored for large-sensor cameras in light-sheet microscopy.
  • Engineered the "Benchtop" mesoSPIM with an expanded field of view and optimized optical components.
  • Integrated features for higher throughput, improved resolution, and compatibility with various tissue clearing techniques.

Main Results:

  • Achieved high spatial resolution of 1.5 µm laterally and 3.3 µm axially across the entire field of view.
  • Enabled magnification up to 20× for detailed imaging.
  • Supported sample sizes ranging from sub-millimeter to several centimeters.
  • Demonstrated compatibility with multiple clearing methods.

Conclusions:

  • The "Benchtop" mesoSPIM represents a significant advancement in accessible, high-performance light-sheet microscopy for large cleared tissues.
  • This next-generation system addresses the growing need for detailed imaging across diverse biological scales.
  • The improved mesoSPIM is poised to accelerate research in neuroscience, developmental biology, pathology, and physics.