triSPIM: light sheet microscopy with isotropic super-resolution.

Area of Science:

• Microscopy and Imaging Technologies
• Biophysics
• Optical Physics

Background:

• Traditional light sheet microscopy faces limitations in achieving isotropic super-resolution, particularly for mesoscopic samples.
• Photodamage remains a critical concern, hindering long-term observation of biological processes.
• Current super-resolution techniques often require complex sample preparation or specialized environments.

Purpose of the Study:

• To propose and theoretically validate a novel three-objective light sheet microscopy geometry.
• To achieve isotropic super-resolution imaging in mesoscopic samples.
• To minimize photodamage while enhancing imaging resolution.

Main Methods:

• Utilizing a three-objective light sheet microscopy setup with skewed lattice light sheet excitation.
• Employing computational fusion of images acquired from two separate lens pairings.
• Investigating the impact of simultaneous coherent excitation through two objectives.

Main Results:

• Demonstrated potential for isotropic super-resolution with a resolution of 120 nm for EGFP imaging.
• Simulations indicate significant resolution enhancement through combined excitation strategies.
• The proposed geometry shows promise in minimizing photodamage compared to conventional methods.

Conclusions:

• The proposed three-objective light sheet microscopy geometry offers a viable path towards isotropic super-resolution.
• Computational image fusion and coherent excitation are key to achieving enhanced resolution.
• This technique has the potential to advance mesoscopic sample imaging with reduced photodamage.