Large-scale tissue clearing (PACT): Technical evaluation and new perspectives in immunofluorescence, histology and ultrastructure

• DOI: 10.1038/srep34331
• Publication Date: 2016-09-29T10:22:03Z
• Authors: Peter H. Neckel; Ulrich Mattheus; Bernhard Hirt; Lothar Just; Andreas F. Mack
• Keywords: Histology; Stain; Eosin

Abstract Novel techniques, like CLARITY and PACT, render large tissue specimens transparent thereby suitable for microscopic analysis. We used these techniques to evaluate their potential in the intestine as an exemplary organ with a complex composition. Immunohistochemistry, light sheet- confocal scanning-microscopy enabled us follow three-dimensional structures, nerve fibers, vessels epithelial barriers throughout entire organ. Moreover, systematic electron study, we analyzed morphology preservation of on ultrastructural level during clearing process. also connect classical histology demonstrate that cleared tissues can be stained Hematoxylin-Eosin Heidenhain’s Azan stain, suggesting use histopathology. These experiments showed neutral pH process results much better ultrastructure standard stainability. Volume changes were monitored quantified course protocol. Additionally, employed technique visualize enteric nervous system barrier post mortem human gut preparations. Our data show high different types supporting its usefulness research diagnosis contribute technical discussion tissue-retention.