Super-speed multiphoton microscopy for mesoscopic volume imaging with ultra-dense sampling beyond Nyquist Limit

Bhaskar Jyoti Borah; Han-Hsiung Chi; Chen-Tung Yen; Chi-Kuang Sun

doi:10.1117/12.2543846

17 February 2020 Super-speed multiphoton microscopy for mesoscopic volume imaging with ultra-dense sampling beyond Nyquist Limit

Bhaskar Jyoti Borah, Han-Hsiung Chi, Chen-Tung Yen, Chi-Kuang Sun

Author Affiliations +

Proceedings Volume 11245, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXVII; 1124515 (2020) https://doi.org/10.1117/12.2543846
Event: SPIE BiOS, 2020, San Francisco, California, United States

Abstract

We developed a nonlinear optical microscopy (NLOM) system with an extended field of view (FOV) of up-to 1.6 mm × 1.6 mm, employing a high numerical aperture (NA) and low magnification objective lens (20×, NA>0.9) maintaining submicron lateral resolution and <2 μm axial resolution, making the system a suitable candidate for high resolution imaging. An acquisition speed of 95 M samples per second was implemented with synchronized sampling of 1 voxel per optical pulse from a femtosecond laser source with a 95 MHz repetition rate, with the ability of scanning a 1.6 × 1.6 × 1.6 mm³ volume, with 12000 × 12000 × 2000 (× 4 channels), i.e., 1.152 Tera-voxels in total, capturing ~2.1 Terabyte of 16-bit raw data with 14-bit resolution in <53 minutes at 0.8 μm Z-steps, and maintaining a Nyquist-exceeded voxel-size, a Nyquist-exceeded volume-scanning speed and a Nyquist-exceeded line-scanning speed of <15 attoliter, >1288 μm³/ ms and >12 mm/ ms, respectively.

Citation Download Citation

Bhaskar Jyoti Borah, Han-Hsiung Chi, Chen-Tung Yen, and Chi-Kuang Sun "Super-speed multiphoton microscopy for mesoscopic volume imaging with ultra-dense sampling beyond Nyquist Limit", Proc. SPIE 11245, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXVII, 1124515 (17 February 2020); https://doi.org/10.1117/12.2543846

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