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- Title:
3D histopathology of human tumours by fast clearing and ultramicroscopy.
- Document type:
- Journal Article
- Author(s):
- Sabdyusheva Litschauer, Inna; Becker, Klaus; Saghafi, Saiedeh; Ballke, Simone; Bollwein, Christine; Foroughipour, Meraaj; Gaugeler, Julia; Foroughipour, Massih; Schavelová, Viktória; László, Viktória; Döme, Balazs; Brostjan, Christine; Weichert, Wilko; Dodt, Hans-Ulrich
- Abstract:
- Here, we describe a novel approach that allows pathologists to three-dimensionally analyse malignant tissues, including the tumour-host tissue interface. Our visualization technique utilizes a combination of ultrafast chemical tissue clearing and light-sheet microscopy to obtain virtual slices and 3D reconstructions of up to multiple centimetre sized tumour resectates. For the clearing of tumours we propose a preparation technique comprising three steps: (a) Fixation and enhancement of tissue autofluorescence with formalin/5-sulfosalicylic acid. (b) Ultrafast active chemical dehydration with 2,2-dimethoxypropane and (c) refractive index matching with dibenzyl ether at up to 56 °C. After clearing, the tumour resectates are imaged. The images are computationally post-processed for contrast enhancement and artefact removal and then 3D reconstructed. Importantly, the sequence a-c is fully reversible, allowing the morphological correlation of one and the same histological structures, once visualized with our novel technique and once visualized by standard H&E- and IHC-staining. After reverting the clearing procedure followed by standard H&E processing, the hallmarks of ductal carcinoma in situ (DCIS) found in the cleared samples could be successfully correlated with the corresponding structures present in H&E and IHC staining. Since the imaging of several thousands of optical sections is a fast process, it is possible to analyse a larger part of the tumour than by mechanical slicing. As this also adds further information about the 3D structure of malignancies, we expect that our technology will become a valuable addition for histological diagnosis in clinical pathology.
- Journal title abbreviation:
- Sci Rep
- Year:
- 2020
- Journal volume:
- 10
- Journal issue:
- 1
- Fulltext / DOI:
- doi:10.1038/s41598-020-71737-w
- Pubmed ID:
- http://view.ncbi.nlm.nih.gov/pubmed/33077794
- Print-ISSN:
- 2045-2322
- TUM Institution:
- Institut für Allgemeine Pathologie und Pathologische Anatomie
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