Three-dimensional virtual histology enabled through cytoplasm-specific X-ray stain for microscopic and nanoscopic computed tomography

Proceedings of the National Academy of Sciences of the United States of America
Madleen BusseFranz Pfeiffer

Abstract

Many histological methods require staining of the cytoplasm, which provides instrumental details for diagnosis. One major limitation is the production of 2D images obtained by destructive preparation of 3D tissue samples. X-ray absorption micro- and nanocomputed tomography (microCT and nanoCT) allows for a nondestructive investigation of a 3D tissue sample, and thus aids to determine regions of interest for further histological examinations. However, application of microCT and nanoCT to biological samples (e.g., biopsies) is limited by the missing contrast within soft tissue, which is important to visualize morphological details. We describe an eosin-based preparation overcoming the challenges of contrast enhancement and selectivity for certain tissues. The eosin-based staining protocol is suitable for whole-organ staining, which then enables high-resolution microCT imaging of whole organs and nanoCT imaging of smaller tissue pieces retrieved from the original sample. Our results demonstrate suitability of the eosin-based staining method for diagnostic screening of 3D tissue samples without impeding further diagnostics through histological methods.

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Citations

Mar 13, 2019·Tissue Engineering. Part a·Liesbet Geris, Ioannis Papantoniou
May 14, 2020·The Analyst·David HartnellMark J Hackett
Jul 7, 2020·Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·Willy KuoVartan Kurtcuoglu
Jun 20, 2018·Proceedings of the National Academy of Sciences of the United States of America·Mareike TöpperwienTim Salditt
Dec 16, 2018·Scientific Reports·Mark MüllerMadleen Busse
Feb 6, 2020·Breast Cancer Research and Treatment·Daniel DiCorpoJames Michaelson
Jul 31, 2019·Annals of Botany·Jan Van den BulckeTom De Mil
Jul 4, 2018·Molecular Imaging and Biology : MIB : the Official Publication of the Academy of Molecular Imaging·J AlbersC Dullin
Mar 3, 2020·Journal of Medical Imaging·Marina EckermannTim Salditt
Nov 5, 2020·Journal of Synchrotron Radiation·Jasper FrohnTim Salditt
Dec 15, 2020·Experimental Eye Research·Sergey Y TkachevMarina S Tkacheva
Dec 11, 2020·Chemistry : a European Journal·Madleen BusseTanja Gulder
Apr 6, 2021·Frontiers in Neuroscience·Paulla Vieira RodriguesMatheus de Castro Fonseca
Apr 28, 2021·Proceedings of the National Academy of Sciences of the United States of America·Daniel KeppelerTobias Moser
Jun 30, 2021·Proceedings of the National Academy of Sciences of the United States of America·Benedikt GeierManuel Liebeke
Jul 4, 2021·International Journal for Parasitology·James D B O'SullivanKathryn J Else
Oct 30, 2021·The British Journal of Radiology·Gerlind SchneiderMatthias Schnabelrauch

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