Live CLEM imaging: an application for yeast cells

Correlative light and electron microscopy (CLEM) is a useful tool for investigating cellular structures at high-resolution. We have extended this technology and developed a new method in which CLEM is combined with live-cell fluorescence imaging, “Live CLEM”. With this method, living cells are first examined by fluorescence microscopy then fixed in situ, and finally the same cells are subjected to electron microscopy. This simple yet powerful method combines temporal and high resolution spatial information. However, due to technical difficulties in immobilizing non-adherent target cells during sample preparation, this method has usually been applied to the study of adherent cells. Here we report a method of live CLEM that can be applied to yeast cells. Since yeast are a powerful tool in molecular genetics, application of this method to the study of yeast should greatly advance our understanding of the molecular basis of the function of cellular structures