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Proceedings Paper

Dual-view inverted selective plane illumination microscopy (diSPIM) with improved background rejection for accurate 3D digital pathology
Author(s): Bihe Hu; Daniel Bolus; J. Quincy Brown
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Paper Abstract

Current gold-standard histopathology for cancerous biopsies is destructive, time consuming, and limited to 2D slices, which do not faithfully represent true 3D tumor micro-morphology. Light sheet microscopy has emerged as a powerful tool for 3D imaging of cancer biospecimens. Here, we utilize the versatile dual-view inverted selective plane illumination microscopy (diSPIM) to render digital histological images of cancer biopsies. Dual-view architecture enabled more isotropic resolution in X, Y, and Z; and different imaging modes, such as adding electronic confocal slit detection (eCSD) or structured illumination (SI), can be used to improve degraded image quality caused by background signal of large, scattering samples. To obtain traditional H&E-like images, we used DRAQ5 and eosin (D&E) staining, with 488nm and 647nm laser illumination, and multi-band filter sets. Here, phantom beads and a D&E stained buccal cell sample have been used to verify our dual-view method. We also show that via dual view imaging and deconvolution, more isotropic resolution has been achieved for optical cleared human prostate sample, providing more accurate quantitation of 3D tumor architecture than was possible with single-view SPIM methods. We demonstrate that the optimized diSPIM delivers more precise analysis of 3D cancer microarchitecture in human prostate biopsy than simpler light sheet microscopy arrangements.

Paper Details

Date Published: 23 February 2018
PDF: 8 pages
Proc. SPIE 10499, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV, 104990Z (23 February 2018); doi: 10.1117/12.2290785
Show Author Affiliations
Bihe Hu, Tulane Univ. (United States)
Daniel Bolus, Tulane Univ. (United States)
J. Quincy Brown, Tulane Univ. (United States)

Published in SPIE Proceedings Vol. 10499:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV
Thomas G. Brown; Carol J. Cogswell; Tony Wilson, Editor(s)

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