Cytometry and fluorescence microscopy / Electron microscopy protocols and methods

Technique / Cytometry and fluorescence microscopy / Electron microscopy

Electron microscopy protocols new recommended protocol
Electron microscopy related protocols by Dr.Christen Harris, Medical College of Georgia. Content: Part A: Tissue Preparation LTP Physiology Protocol Vibroslicer VT 1000S Slice Dissection and Vibratoming in Agarose Stoelting Tissue Chopper ...

Gold labeling technique for electron microscopic identification and location of proteins protocol
Written by Maria Schnos, Institute for Molecular Virology, University of Wisconsin - Madison. The method that has previously been used to gold-label proteins at the molecular level uses the following strategy (RecA protein bound to DNA will be used as an e ...

Immunogold labeling for two or more antigens protocol
This is the favorite electron microscopic immunolabeling protocol in the laboratory of Dr. G. V Childs at the University of Texas Medical Branch . The purpose of this protocol is to allow labeling for two (or more) antigens with sensitive, small protein-A conj ...

Standard Glutaraldehyde Fixation new protocol
Standard Glutaraldehyde Fixation Protocl for TEM of Tissue (from EMSA interdisciplinary center at the Universiteit Utrecht). ...

Plasma Cleaner Operation Manual protocol
For optimal imaging and microanalysis in electron microscopy, it is imperative to have a clean, wellprepared specimen. This is the manual for Model 1020 Plasma Cleaner. (from EMSA interdisciplinary center at the Universiteit Utrecht). ...

Cryo-immunogold electron
(Curr Protoc Cell Biol. 2006 Apr;Chapter 4:Unit 4.7)
microscopy.Peters PJ, Bos E, Griekspoor A.
Netherlands Cancer Institute, Amsterdam, The Netherlands.

This unit describes subcellular localization of proteins/antigens using high-resolution cryo-immunogold electron microscopy, which allows study of topological biochemistry at the ultrastructural level. This is the most sensitive procedure for immunodetection of antigens on ultrathin sections prepared from chemically fixed cells or tissues, because aldehyde fixation is the only denaturation step. The omission of harsh organic solvents (such as those used for plastic embedding) ensures better preservation of protein antigenicity. Support protocols describe how to embed fixed material in gelatin, cryosection, and mount the sections on Formvar-coated grids. This unit is accompanied by eleven videos that illustrate many of the procedures used in this unit.

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Last update:

09-May-2008 11:46 am

Related new papers and reviews

Electron microscopy of the early Caenorhabditis elegans embryo.

The early Caenorhabditis elegans embryo is currently a popular model system to study centrosome assembly, kinetochore organization, spindle formation, and cellular polarization. Here, we present and review methods for routine electron microscopy and 3D analysis of the early C. elegans embryo. The first method uses laser-induced chemical fixation to preserve the fine structure of isolated embryos. This approach takes advantage of time-resolved fixation to arrest development at specific stages. The second method uses high-pressure freezing of whole worms followed by freeze-substitution (HPF-FS) for ultrastructural analysis. This technique allows staging of developing early embryos within the worm uterus, and has the advantage of superior sample preservation required for high-resolution 3D reconstruction. The third method uses a correlative approach to stage isolated, single embryos by light microscopy followed by HPF-FS and electron tomography. This procedure combines the advantages of time-resolved fixation and superior ultrastructural preservation by high-pressure freezing and allows a higher throughput electron microscopic analysis. The advantages and disadvantages of these methods for different applications are discussed.