Altair-LSFM A High-Resolution, Easy-to-Build Light-Sheet Microscope for Sub-Cellular Imaging

Area of Science:

• Biophysics
• Microscopy
• Cell Biology

Background:

• Existing open-source light-sheet microscopes (e.g., mesoSPIM, OpenSPIM) are suitable for large specimens but lack resolution for sub-cellular details.
• Latice Light-Sheet Microscopy (LLSM) offers high resolution but can be complex to align and maintain in open-source versions.

Purpose of the Study:

• To develop a high-resolution, open-source light-sheet microscope optimized for sub-cellular imaging.
• To simplify the assembly and alignment process for researchers without specialized expertise.

Main Methods:

• Designed Altair-LSFM with an optimized optical pathway and a custom baseplate for simplified assembly.
• Integrated streamlined optoelectronics and optomechanics.
• Utilized open-source software (navigate) for seamless operation.

Main Results:

• Achieved lateral and axial resolutions of ~235 nm and ~350 nm, respectively, over a 266-micron field of view (post-deconvolution).
• Successfully imaged sub-diffraction fluorescent nanospheres.
• Visualized fine sub-cellular structures in mammalian cells (microtubules, actin, nuclei, Golgi).
• Demonstrated live-cell imaging of dynamic cytoskeletal elements in migrating cells.

Conclusions:

• Altair-LSFM provides a user-friendly, high-resolution platform for sub-cellular imaging.
• The system overcomes the limitations of existing open-source microscopes for detailed cellular and live-cell studies.
• Enables advanced research into cellular architecture and dynamics.