title: Correlative stochastic optical reconstruction microscopy and electron microscopy.
abstract: Correlative fluorescence light microscopy and electron microscopy allows the imaging of spatial distributions of specific biomolecules in the context of cellular ultrastructure. Recent development of super-resolution fluorescence microscopy allows the location of molecules to be determined with nanometer-scale spatial resolution. However, correlative super-resolution fluorescence microscopy and electron microscopy (EM) still remains challenging because the optimal specimen preparation and imaging conditions for super-resolution fluorescence microscopy and EM are often not compatible. Here, we have developed several experiment protocols for correlative stochastic optical reconstruction microscopy (STORM) and EM methods, both for un-embedded samples by applying EM-specific sample preparations after STORM imaging and for embedded and sectioned samples by optimizing the fluorescence under EM fixation, staining and embedding conditions. We demonstrated these methods using a variety of cellular targets.
authors: Howard Hughes Medical Institute, Cambridge, Massachusetts, United States of America; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, United States of America.
authors: Doory
authors: D
authors: Kim
authors: National Center for Microscopy and Imaging Research, Center for Research in Biological Systems, Department of Neurosciences, University of California San Diego, La Jolla, California, United States of America.
authors: Thomas J
authors: TJ
authors: Deerinck
authors: Howard Hughes Medical Institute, Cambridge, Massachusetts, United States of America; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, United States of America.
authors: Yaron M
authors: YM
authors: Sigal
authors: Center for Brain Science, Harvard University, Cambridge, Massachusetts, United States of America.
authors: Hazen P
authors: HP
authors: Babcock
authors: National Center for Microscopy and Imaging Research, Center for Research in Biological Systems, Department of Neurosciences, University of California San Diego, La Jolla, California, United States of America.
authors: Mark H
authors: MH
authors: Ellisman
authors: Howard Hughes Medical Institute, Cambridge, Massachusetts, United States of America; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, United States of America; Center for Brain Science, Harvard University, Cambridge, Massachusetts, United States of America; Department of Physics, Harvard University, Cambridge, Massachusetts, United States of America.
authors: Xiaowei
authors: X
authors: Zhuang
mesh_terms: D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D004847:Epithelial Cells / Q000648:ultrastructure; D006801:Humans; D008854:Microscopy, Electron / Q000295:instrumentation / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D008867:Microtomy; D008870:Microtubules / Q000648:ultrastructure; D009975:Orthomyxoviridae / Q000648:ultrastructure; D013194:Staining and Labeling / Q000379:methods*; D013269:Stochastic Processes; D016610:Tissue Embedding; D057074:Virus Release / Q000502:physiology