FITC-Dextran tracers in microcirculatory and permeability studies using combined fluorescence stereo microscopy, fluorescence light microscopy and electron microscopy

1981 ◽  
Vol 71 (2) ◽  
pp. 209-233 ◽  
Author(s):  
N. Thorball
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Fluorescence Light Microscopy ◽  
Fluorescence Light ◽  
Permeability Studies

Advances in Correlative Single-Molecule Localization Microscopy and Electron Microscopy

2015 ◽  
Vol 1 (1) ◽  
Author(s):  
Ulrike Endesfelder
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Single Molecule ◽  
Light And Electron Microscopy ◽  
Fluorescence Labeling ◽  
Localization Microscopy ◽  
Fluorescence Light Microscopy ◽  
Fluorescence Light ◽  
And Function ◽  
Single Molecule Localization Microscopy

AbstractDuring the last few decades, correlative fluorescence light and electron microscopy (FLM-EM) has gained increased interest in the life sciences concomitant with the advent of fluorescence light microscopy. It has become, accompanied by numerous developments in both techniques, an important tool to study bio-cellular structure and function as it combines the specificity of fluorescence labeling with the high structural resolution and cellular context information given by the EM images. Having the recently introduced single-molecule localization microscopy techniques (SMLM) at hand, FLM-EM can now make use of improved fluorescence light microscopy resolution, single-molecule sensitivity and quantification strategies. Here, currently used methods for correlative SMLM and EM including the special requirements in sample preparation and imaging routines are summarized and an outlook on remaining challenges concerning methods and instrumentation is provided.

A simple preparation method for CLEM using pre-embedding immunohistochemistry with a novel fluorescent probe and stable embedding resin

Microscopy ◽  
2021 ◽  
Author(s):  
Takaaki Kanemaru ◽  
Teruyoshi Kondo ◽  
Kei-ichiro Nakamura ◽  
Hiroyuki Morimoto ◽  
Kentaro Nishi ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Electron Beam ◽  
High Resolution ◽  
Light Microscopy ◽  
Fluorescent Probe ◽  
Region Of Interest ◽  
Wide Field ◽  
Lr White ◽  
Fluorescence Light Microscopy ◽  
Fluorescence Light

Abstract Correlative light and electron microscopy (CLEM) is an excellent approach for examining the cellular localization of biomolecules. Here, we developed a simple method for CLEM by combining pre-embedding immunohistochemistry with a novel fluorescent probe, namely Fluolid NS Orange, and an embedding resin called ʻDurcupan™ʼ. Specimens were embedded in Durcupan™ or LR White after immunolabeling and post-fixation using glutaraldehyde and osmium tetroxide. Next, ultrathin sections were prepared on a finder grid with navigation markers. The section of the specimen embedded in Durcupan™ was found to be more stable against electron beam irradiation than specimens embedded in LR White. A fluorescence light microscopy image and a transmission electron microscopy (TEM) image, at wide-field, and low magnification, were independently obtained with the same ultrathin section. Using the three corners between finder grid bars as landmarks, fluorescence light microscopy images were superimposed with wide-field, low-magnification TEM images to identify the region of interest, which was subsequently enlarged to ascertain cellular structures localized beneath fluorescent signals. However, the enlarged TEM images appeared blurred, and fluorescence signals had a hazy appearance. To resolve this, the enlarged TEM images were replaced by high-resolution TEM images focused directly on the region of interest, thereby facilitating the collection of high-resolution CLEM images. The simple sample processing method for CLEM using osmium-resistant Fluolid NS Orange and electron beam damage-resistant Durcupan™ allowed the determination of the precise localization of fluorescence signals at subcellular levels.

scholarly journals Micro axial tomography: A miniaturized, versatile stage device to overcome resolution anisotropy in fluorescence light microscopy

2011 ◽  
Vol 82 (9) ◽  
pp. 093701 ◽  
Author(s):  
Florian Staier ◽  
Heinz Eipel ◽  
Petr Matula ◽  
Alexei V. Evsikov ◽  
Michal Kozubek ◽  
...  
Keyword(s):  
Light Microscopy ◽  
Axial Tomography ◽  
Fluorescence Light Microscopy ◽  
Fluorescence Light

Monitoring and quantifying of oil migration in cocoa butter using a flatbed scanner and fluorescence light microscopy

2005 ◽  
Vol 38 (10) ◽  
pp. 1189-1197 ◽  
Author(s):  
Stéphanie Marty ◽  
Ken Baker ◽  
Elena Dibildox-Alvarado ◽  
Juliana Neves Rodrigues ◽  
Alejandro G. Marangoni
Keyword(s):  
Cocoa Butter ◽  
Light Microscopy ◽  
Flatbed Scanner ◽  
Oil Migration ◽  
Fluorescence Light Microscopy ◽  
Fluorescence Light

scholarly journals COMBO-FISH: A Versatile Tool Beyond Standard FISH to Study Chromatin Organization by Fluorescence Light Microscopy

OBM Genetics ◽  
2018 ◽  
Vol 3 (1) ◽  
pp. 1-1 ◽  
Author(s):  
Jin-Ho Lee ◽  
◽  
Florence Laure Djikimi Tchetgna ◽  
Matthias Krufczik ◽  
Eberhard Schmitt ◽  
...  
Keyword(s):  
Light Microscopy ◽  
Chromatin Organization ◽  
Fluorescence Light Microscopy ◽  
Versatile Tool ◽  
Fluorescence Light

scholarly journals CorRelator: An interactive and flexible toolkit for high-precision cryo-correlative light and electron microscopy

2020 ◽  
Author(s):  
Jie E. Yang ◽  
Matthew R. Larson ◽  
Bryan S. Sibert ◽  
Samantha Shrum ◽  
Elizabeth R. Wright
Keyword(s):  
Electron Microscopy ◽  
Light And Electron Microscopy ◽  
Cryo Electron Microscopy ◽  
Cellular Environment ◽  
Transmission Electron ◽  
Desktop Application ◽  
Fluorescence Light Microscopy ◽  
Cross Platform ◽  
Fluorescence Light ◽  
Correlation Process

AbstractCryo-correlative light and electron microscopy (CLEM) is a technique that uses the spatiotemporal cues from fluorescence light microscopy (FLM) to investigate the high-resolution ultrastructure of biological samples by cryo-electron microscopy (cryo-EM). Cryo-CLEM provides advantages for identifying and distinguishing fluorescently labeled proteins, macromolecular complexes, and organelles from the cellular environment. Challenges remain on how correlation workflows and software tools are implemented on different microscope platforms to support microscopy-driven structural studies. Here, we present an open-source desktop application tool, CorRelator, to bridge between cryo-FLM and cryo-EM/ET data collection instruments. CorRelator was designed to be flexible for both on-the-fly and post-acquisition correlation schemes. The CorRelator workflow is easily adapted to any fluorescence and transmission electron microscope (TEM) system configuration. CorRelator was benchmarked under cryogenic and ambient temperature conditions using several FLM and TEM instruments, demonstrating that CorRelator is a rapid and efficient application for image and position registration in CLEM studies. CorRelator is a cross-platform software featuring an intuitive Graphical User Interface (GUI) that guides the user through the correlation process. CorRelator source code is available at: https://github.com/wright-cemrc-projects/corr.

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