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 (PCa). 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 CK8-18 resulted in comprehensive 3D imaging of benign peripheral and transition prostate zones, as well as individual PCa 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.
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 (PCa). 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 CK8-18 resulted in comprehensive 3D imaging of benign peripheral and transition prostate zones, as well as individual PCa 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.
Authors: Michael C Haffner; Wilbert Zwart; Martine P Roudier; Lawrence D True; William G Nelson; Jonathan I Epstein; Angelo M De Marzo; Peter S Nelson; Srinivasan Yegnasubramanian Journal: Nat Rev Urol Date: 2020-12-16 Impact factor: 14.432
Authors: Floris J Voskuil; Jasper Vonk; Bert van der Vegt; Schelto Kruijff; Vasilis Ntziachristos; Pieter J van der Zaag; Max J H Witjes; Gooitzen M van Dam Journal: Nat Biomed Eng Date: 2021-11-08 Impact factor: 25.671
Authors: Weisi Xie; Nicholas P Reder; Can Koyuncu; Patrick Leo; Sarah Hawley; Hongyi Huang; Chenyi Mao; Nadia Postupna; Soyoung Kang; Robert Serafin; Gan Gao; Qinghua Han; Kevin W Bishop; Lindsey A Barner; Pingfu Fu; Jonathan L Wright; C Dirk Keene; Joshua C Vaughan; Andrew Janowczyk; Adam K Glaser; Anant Madabhushi; Lawrence D True; Jonathan T C Liu Journal: Cancer Res Date: 2021-12-01 Impact factor: 13.312
Authors: Jonathan T C Liu; Adam K Glaser; Kaustav Bera; Lawrence D True; Nicholas P Reder; Kevin W Eliceiri; Anant Madabhushi Journal: Nat Biomed Eng Date: 2021-02-15 Impact factor: 25.671