![]() ![]() ![]() The mutant protein is retained in the endoplasmic reticulum (ER) and could therefore alter ER function and morphology. We use FLIPPER to quantify cellular morphology at the EM level in cells expressing a normal and disease-causing point-mutant cell-surface protein called EpCAM (epithelial cell adhesion molecule). The use of FLIPPER is straightforward and because the module is completely genetically encoded, cells can be optimally prepared for EM examination. FLIPPER consists of a fluorescent protein (cyan, green, orange, or red) for LM visualization, fused to a peroxidase allowing visualization of targets at the EM level. We present a new genetically encoded probe for CLEM, named “FLIPPER”, which facilitates quantitative analysis of ultrastructural features in cells. Even when combining light microscopy (LM) with EM (correlated LM and EM: CLEM) to find areas of interest, the labeling of molecules is still a challenge. However, EM is typically restricted to small fields of view at high magnification this makes quantifying events in multiple large-area sample sections extremely difficult. Ultrastructural examination of cells and tissues by electron microscopy (EM) yields detailed information on subcellular structures. ![]()
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