Live CLEM imaging to analyze nuclear structures at high resolution

Abstract

Fluorescence microscopy (FM) and electron microscopy (EM) are powerful tools for observing molecular components in cells. FM can provide temporal information about cellular proteins and structures in living cells. EM provides nanometer resolution images of cellular structures in fi xed cells. We have combined FM and EM to develop a new method of correlative light and electron microscopy (CLEM), called “Live CLEM.” In this method, the dynamic behavior of specifi c molecules of interest is fi rst observed in living cells using fl uorescence microscopy (FM) and then cellular structures in the same cell are observed using electron microscopy (EM). Following image acquisition, FM and EM images are compared to enable the fl uorescent images to be correlated with the high-resolution images of cellular structures obtained using EM. As this method enables analysis of dynamic events involving specifi c molecules of interest in the context of specifi c cellular structures at high resolution, it is useful for the study of nuclear structures including nuclear bodies. Here we describe Live CLEM that can be applied to the study of nuclear structures in mammalian cells.