DRAQ5 and Eosin (‘D&E’) as an Analog to Hematoxylin and Eosin for Rapid Fluorescence Histology of Fresh Tissues
AuthorElfer, Katherine N.
Sholl, Andrew B.
Tulman, David B.
Mandava, Sree H.
Lee, Benjamin R.
Brown, J. Quincy
AffiliationUniv Arizona Hlth Sci, Dept Surg, Div Urol
MetadataShow full item record
PublisherPUBLIC LIBRARY SCIENCE
CitationDRAQ5 and Eosin (‘D&E’) as an Analog to Hematoxylin and Eosin for Rapid Fluorescence Histology of Fresh Tissues 2016, 11 (10):e0165530 PLOS ONE
Rights© 2016 Elfer et al. This is an open access article distributed under the terms of the Creative Commons Attribution License.
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AbstractReal-time on-site histopathology review of biopsy tissues at the point-of-procedure has great potential for significant clinical value and improved patient care. For instance, on-site review can aid in rapid screening of diagnostic biopsies to reduce false-negative results, or in quantitative assessment of biospecimen quality to increase the efficacy of downstream laboratory and histopathology analysis. However, the only currently available rapid pathology method, frozen section analysis (FSA), is too time-and labor-intensive for use in screening large quantities of biopsy tissues and is too destructive for maximum tissue conservation in multiple small needle core biopsies. In this work we demonstrate the spectrally-compatible combination of the nuclear stain DRAQ5 and the anionic counterstain eosin as a dual-component fluorescent staining analog to hematoxylin and eosin intended for use on fresh, unsectioned tissues. Combined with optical sectioning fluorescence microscopy and pseudo-coloring algorithms, DRAQ5 and eosin ("D&E") enables very fast, non-destructive psuedohistological imaging of tissues at the point-of-acquisition with minimal tissue handling and processing. D&E was validated against H&E on a one-to-one basis on formalin-fixed paraffin-embedded and frozen section tissues of various human organs using standard epi-fluorescence microscopy, demonstrating high fidelity of the staining mechanism as an H&E analog. The method was then applied to fresh, whole 18G renal needle core biopsies and large needle core prostate biospecimen biopsies using fluorescence structured illumination optical sectioning microscopy. We demonstrate the ability to obtain high-resolution histology-like images of unsectioned, fresh tissues similar to subsequent H&E staining of the tissue. The application of D&E does not interfere with subsequent standard-of-care H&E staining and imaging, preserving the integrity of the tissue for thorough downstream analysis. These results indicate that this dual-stain pseudocoloring method could provide a real-time histology-like image at the time of acquisition and valuable objective tissue analysis for the clinician at the time of service.
NoteOpen Access Journal.
VersionFinal published version
SponsorsNational Cancer Institute of the National Institutes of Health [R21 CA159936]; Innovative Molecular Analysis Technologies (IMAT) program of the National Cancer Institute [R33 CM 96457]; Tulane University School of Science and Engineering; National Science Foundation Graduate Research Fellowship Program; National Science Foundation IGERT Fellowship [DGE-1144646]