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Three‐dimensional microscopic analysis of clinical prostate specimens
Author(s) -
Royen Martin E,
Verhoef Esther I,
Kweldam Charlotte F,
Cappellen Wiggert A,
Kremers GertJan,
Houtsmuller Adriaan B,
Leenders Geert J L H
Publication year - 2016
Publication title -
histopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.626
H-Index - 124
eISSN - 1365-2559
pISSN - 0309-0167
DOI - 10.1111/his.13022
Subject(s) - prostate , prostate cancer , pathology , context (archaeology) , staining , cytokeratin , microscopy , medicine , biomedical engineering , immunohistochemistry , biology , cancer , paleontology
Aims Microscopic evaluation of prostate specimens for both clinical and research purposes is generally performed on 5‐μm‐thick tissue sections. Because cross‐sections give a two‐dimensional (2D) representation, little is known about the actual underlying three‐dimensional (3D) architectural features of benign prostate tissue and prostate cancer ( PC a). The aim of this study was to show that a combination of tissue‐clearing protocols and confocal microscopy can successfully be applied to investigate the 3D architecture of human prostate tissue. Methods and results Optical clearing of intact fresh and formalin‐fixed paraffin‐embedded ( FFPE ) clinical prostate specimens allowed us to visualize tissue structures up to a depth of 800 μm, whereas, in uncleared tissue, detection of fluorescence was only possible up to 70 μm. Fluorescent labelling with a general nuclear dye and antibodies against cytokeratin ( CK ) 5 and CK 8‐18 resulted in comprehensive 3D imaging of benign peripheral and transition prostate zones, as well as individual PC a growth patterns. After staining, clearing, and imaging, samples could still be processed for 2D (immuno)histochemical staining and DNA analysis, enabling additional molecular and diagnostic characterization of small tissue specimens. Conclusions In conclusion, the applicability of 3D imaging to archival FFPE and fresh clinical specimens offers unlimited opportunities to study clinical and biological topics of interest in their actual 3D context.