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Ultra-fast multiphoton microscopy for diagnostic histology with polygon, stage-scanning, and ultra-high repetition laser (Conference Presentation)
Author(s): Richard Torres; Eben Olson; Michael J. Levene
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Paper Abstract

Methods for imaging of excised tissue specimens that obviate manual and time-consuming histology processing steps including wax-embedding, sectioning, and separate staining hold considerable promise for improving our ability to render time- and cost-efficient diagnoses. They also could potentiate the clinical adoption of machine learning tools dependent on routine production of digital histology data. A known limitation of point scanning-based ex vivo imaging technologies that precludes ready adoption for clinical use is imaging speed. We have recently described the development of an approach for multiphoton microscopy capable of imaging un-sectioned and un-embedded human tissue samples at millimeter depths with sufficient quality for primary diagnostic interpretation and at speeds more than 30-times faster than a traditional galvanometer-based system. Incorporation of a polygonal mirror combined with stage scanning in an optically efficient geometry yielded significant speed gains, beyond those possible with resonant galvanometers, and without sacrificing image quality. We hypothesized that further gains in speed, with maintenance of image quality, would be attainable by incorporating pulsed laser excitation with a repetition rate beyond the typical 80 MHz speed of standard Ti-Sapphire lasers. In this analysis we describe the use of a newly produced 250 MHz ultra-fast laser in a polygon-based microscope with stage scanning and demonstrate multiphoton human tissue imaging at speeds on par with the fastest whole slide imaging systems and with resolution, contrast, and coloration that matches physical slides but without any of the principal artifacts associated with slide scanning.

Paper Details

Date Published: 4 March 2019
Proc. SPIE 10889, High-Speed Biomedical Imaging and Spectroscopy IV, 108890B (4 March 2019); doi: 10.1117/12.2507986
Show Author Affiliations
Richard Torres, Yale School of Medicine (United States)
Eben Olson, Yale School of Medicine (United States)
Michael J. Levene, Applikate Technologies (United States)

Published in SPIE Proceedings Vol. 10889:
High-Speed Biomedical Imaging and Spectroscopy IV
Kevin K. Tsia; Keisuke Goda, Editor(s)

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