Simple correlative imaging of proteins via in-situ silver intensification

Abstract

With the advances in microscopy, studies on correlative imaging methods, such as correlative light and electron microscopy (CLEM), have been studied. However, CLEM techniques are limited when using specific microscopes, or due to problem in sample preparation or probes itself. Herein, we propose a new CLEM technique based on gold-silver immunostaining, Dehydration-correlative microscopy (DeCom). Dehydration under ambient conditions allows simple and quick correlative imaging. Various analyses, including STORM, revealed that correlative imaging is possible despite sample dehydration under ambient conditions, and the change of the sample according to the growth of silver was confirmed. Additionally, multicolor imaging of cytoskeletal proteins, and microtubule severing protein with tubulin showed the possibility that DeCom can be used in several studies. As a result, DeCom has fewer instrumental requirements than SIM, super-resolution optical techniques, or other CLEM techniques, and will be particularly useful as a technique that provide high resolution images in a short time using SEM in the studies of cytoskeleton and cytoskeleton-related proteins.