Seminar: "Ultra-nanostructure Expansion Microscopy of Paraffin Embedded Mouse Testis Tissue Allows Enhanced Detection of Spatially Restricted Somatic and Germ Cell Protein" - Ashley Di Agostino
Endocrinology and Animal Biosciences Seminar
Ashley Di Agostino
PhD Student
Graduate Program in Endocrinology and Animal Biosciences, Rutgers
Expansion microscopy (ExM) has emerged as a powerful tool to study subcellular structures not readily visualized by standard immunofluorescence. By leveraging the hydrophilic properties of acrylamide hydrogels, ExM can expand embedded tissues up to nine times the original size. As a result, near-super-resolution imaging can be achieved using standard light microscopy. This feature makes ExM of special utility in cell systems reliant on refined subcellular spatial events such as the mammalian male germ cell. Despite this, a protocol for the application of ExM to fixed mammalian testes has not been developed and thoroughly evaluated. Further, best-practice approaches for expansion factor (EF) calculations remain undefined. This seminar describes and tests a modified ExM protocol applied to formalin-fixed, paraffin-embedded mouse testis tissue. Using this approach, we quantify tissue and cell-level expansion parameters and define the localization of spatially restricted proteins important for somatic cell function, meiosis, and spermiogenesis at the sub-micron scale.