FluoroNanogold: an important probe for correlative microscopy
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REVIEW
FluoroNanogold: an important probe for correlative microscopy Toshihiro Takizawa 1 & Richard D. Powell 2 & James F. Hainfeld 2 & John M. Robinson 3
Received: 10 July 2015 / Accepted: 24 July 2015 # Springer-Verlag Berlin Heidelberg 2015
Abstract Correlative microscopy is a powerful imaging approach that refers to observing the same exact structures within a specimen by two or more imaging modalities. In biological samples, this typically means examining the same subcellular feature with different imaging methods. Correlative microscopy is not restricted to the domains of fluorescence microscopy and electron microscopy; however, currently, most correlative microscopy studies combine these two methods, and in this review, we will focus on the use of fluorescence and electron microscopy. Successful correlative fluorescence and electron microscopy requires probes, or reporter systems, from which useful information can be obtained with each of the imaging modalities employed. The bi-functional immunolabeling reagent, FluoroNanogold, is one such probe that provides robust signals in both fluorescence and electron microscopy. It consists of a gold cluster compound that is visualized by electron microscopy and a covalently attached fluorophore that is visualized by fluorescence microscopy. FluoroNanogold has been an extremely useful labeling reagent in correlative microscopy studies. In this report, we present an overview of research using this unique probe.
* Richard D. Powell [email protected] 1
Department of Molecular Anatomy, Nippon Medical School, Tokyo, Japan
2
Nanoprobes, Incorporated, 95 Horseblock Road, Unit 1, Yaphank, NY 11980-9710, USA
3
Department of Physiology and Cell Biology, Ohio State University, Columbus, OH 43210, USA
Keywords FluoroNanogold . Nanogold . Correlative microscopy . Ultrathin cryosections . Immunocytochemistry
Introduction In this review, the term correlative microscopy refers to examining the same exact structures within a biological specimen (e.g., a sub-cellular feature) using two, or more, imaging modalities. The goal of correlative microscopy is to provide greater information than can be provided by any individual imaging method. A number of biological questions have been approached using different aspects of correlative microscopy. However, combining fluorescence and electron microscopy remains the most common form of correlative microscopy. Different aspects of correlative fluorescence and electron microscopy have been discussed in a number of excellent reviews and articles; the reader is referred to these for a broader discussion of correlative microscopy (e.g., [5, 10, 24, 40, 42, 58, 67, 68, 70, 83, 84, 92, 109]). Herein, we will focus on correlative fluorescence and electron microscopy using the bifunctional probe, FluoroNanogold (FNG). Additionally, we will discuss the merits of using ultrathin cryosections, particularly of tissues, as the labeling substrate for correlative microscopy. Several fluorescent markers have been used in correlative microscopy or
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