Immunoelectron Microscopy of Keratinocytes: an Improved Protocol for Flat Embedding of Cultured Cells Using Lowicryl K4M

Abstract

Abstract In cell culture models various and distinct manipulations can be performed in a defined environment. This makes cell cultures very popular for immunohistochemical studies. Such studies are mostly performed at the light microscopic level where fluorescent probes and confocal microscopy provide detailed insights into the distribution and localization of antigens inside cells. In many such cases further studies at the electron microscopic level would give additional information and an often more detailed view. As compared to light microscopy not only is the resolution much higher, but also and even more importantly labeling information is completed with a wealth of cytological details. This said it is surprising that studies on cell cultures using immunoelectron microscopy are rarely seen. Even more rare are reports in which monolayer cells are fixed and processed in situ without prior trypsinization. Such studies are essential though when cytoarchitecture is of importance or especially when cell-cell adhesion is an issue.