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Proceedings Paper

Cell counting in whole mount tissue volumes using expansion OCT (Conference Presentation)
Author(s): Yehe Liu; Shi Gu; Michiko Watanabe; Andrew M. Rollins; Michael W. Jenkins

Paper Abstract

Abnormal cell proliferation and migration during heart development can lead to severe congenital heart defects (CHDs). Studying the spatial distribution of cells during embryonic development helps our understanding of how the heart develops and the etiology of certain CHDs. However, imaging large groups of single cells in intact tissue volumes is challenging. No current technique can accomplish this task in both a time-efficient and cost-effective manner. OCT has potential with its large field of view and micron-scale resolution, but even the highest resolution OCT systems have poor contrast for counting cells and have a small field of view compared to conventional OCT. We propose using a conventional OCT system and processing the sample to enhance cellular contrast. Inspired by the recently developed Expansion Microscopy, we permeated whole-mount embryonic tissue with a superabsorbent monomer solution and polymerized into a hydrogel. When hydrated in DI water, the tissue-hydrogel complex was uniformly enlarged (~5X in all dimensions) without distorting the microscopic structure. This had a twofold effect: it increased the resolution by a factor of 5 and decreased scattering, which allowed us to resolve cellular level features deep in the tissue with high contrast using conventional OCT. We noted that cell nuclei caused significantly more backscattering than the other subcellular structures after expansion. Based on this property, we were able to distinguish individual cell nuclei, and thus count cells, in expanded OCT images with simple intensity thresholding. We demonstrate the technique with embryonic quail hearts at various developmental stages.

Paper Details

Date Published: 19 April 2017
PDF: 1 pages
Proc. SPIE 10043, Diagnosis and Treatment of Diseases in the Breast and Reproductive System, 100430R (19 April 2017); doi: 10.1117/12.2253162
Show Author Affiliations
Yehe Liu, Case Western Reserve Univ. (United States)
Shi Gu, Case Western Reserve Univ. (United States)
Michiko Watanabe, Case Western Reserve Univ. (United States)
Andrew M. Rollins, Case Western Reserve Univ. (United States)
Michael W. Jenkins, Case Western Reserve Univ. (United States)

Published in SPIE Proceedings Vol. 10043:
Diagnosis and Treatment of Diseases in the Breast and Reproductive System
Melissa C. Skala; Paul J. Campagnola, Editor(s)

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