EnzMet™: An Enzymatic Metallography Reagent for Accurately Delineating Neuronal Boundaries for Segmenting Gap Junction-Coupled Neurons in their Three-dimensional Space

Torres-Vazquez, Irma; Serrano-Vélez, José L.; Rosa‐Molinar, Eduardo; Orange, François; Guinel, Maxime J.-F.; Koutis, Ioannis; Wolf, Johannes; Laimer, C.; Joshi, Vishwas N.; Powell, Rick

doi:10.1017/s1431927612005156

Cited by 1 publication

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“…The development of serial section methods for electron microscopy, such as serial blockface SEM and focused ion beam (FIB) SEM, provides another opportunity for correlation between electron and light microscopic methods. While these methods do not currently achieve sufficient resolution to visualize individual NG particles, X-ray microscopy has been used as a guide to correlate regions of interest found by fluorescence with subsequent serial block face SEM analysis [7], while a combination of fluorescence backfilling with fluorescently labeled biocytin and metallographic contrasting using enzyme metallography [53,107] has been used to correlate fluorescence with serial block face SEM. A fundamental challenge with these methods is the development of probes small enough to penetrate the thick specimens typically analyzed by such methods, yet producing sufficient EM signal for detection.…”

Section: Future Directionsmentioning

confidence: 99%

FluoroNanogold: an important probe for correlative microscopy

et al. 2015

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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 sub-cellular 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.

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Section: Future Directionsmentioning

confidence: 99%