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Proceedings Paper

Super-speed multiphoton microscopy for mesoscopic volume imaging with ultra-dense sampling beyond Nyquist Limit
Author(s): Bhaskar Jyoti Borah; Han-Hsiung Chi; Chen-Tung Yen; Chi-Kuang Sun
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Paper 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.

Paper Details

Date Published: 17 February 2020
PDF: 6 pages
Proc. SPIE 11245, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXVII, 1124515 (17 February 2020); doi: 10.1117/12.2543846
Show Author Affiliations
Bhaskar Jyoti Borah, National Taiwan Univ. (Taiwan)
Han-Hsiung Chi, National Taiwan Univ. (Taiwan)
Chen-Tung Yen, National Taiwan Univ. (Taiwan)
Chi-Kuang Sun, National Taiwan Univ. (Taiwan)
Academia Sinica (Taiwan)

Published in SPIE Proceedings Vol. 11245:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXVII
Thomas G. Brown; Tony Wilson; Laura Waller, Editor(s)

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