In Situ Embedding of Cell Monolayers Cultured on Plastic Surfaces for Electron Microscopy

1991 ◽  
Vol 66 (5) ◽  
pp. 269-272 ◽  
Author(s):  
A. Beatrice Murray ◽  
Helga Schulze ◽  
Elisabeth Blauw
Keyword(s):  
Electron Microscopy ◽  
Cell Monolayers ◽  
Plastic Surfaces ◽  
In Situ Embedding

scholarly journals Simplified in situ preparation of cultured cell monolayers for electron microscopy.

1980 ◽  
Vol 28 (2) ◽  
pp. 178-180 ◽  
Author(s):  
K Kawamoto ◽  
A Hirano ◽  
F Herz
Keyword(s):  
Electron Microscopy ◽  
Cultured Cells ◽  
Growth Surface ◽  
Cultured Cell ◽  
Cell Monolayers ◽  
In Situ Preparation

The use of xylene for the easy separation of cultured cells embedded in situ from their plastic growth surface is described. This step simplifies the preparation of cell monolayers for electron microscopy.

In Situ Embedding of Cell Monolayers on Untreated Glass Surfaces for Vertical and Horizontal Ultramicrotomy.

1980 ◽  
Vol 38 ◽  
pp. 644-645
Author(s):  
Kai Chien
Keyword(s):  
Organic Solvent ◽  
Light Microscopy ◽  
Cell Monolayer ◽  
Glass Slide ◽  
Plastic Substrates ◽  
Cell Monolayers ◽  
Glass Surfaces ◽  
In Situ Embedding ◽  
The Ideal

Untreated glass is the ideal supporting substrate for cell cul¬ture growth since it is extremely flat and smooth, transparent and insoluable in organic solvent. However, difficulties have been encountered in removing polymerized epoxy resin from a glass surface following in situ cell monolayer embedment. Vari¬ous techniques have been made to grow cell cultures on either coated glass surfaces or plastic substrates. The purpose of this abstract is to describe a heat separation technique which when used together with a newly designed embedding mold allows cell monolayers to be transferred from untreated coverglass or glass slide to pre-shaped tissue blocks. The resulting tissue block can be easily separated and used directly for orientation light microscopy prior to ultramicrotomy.

The use of Lab-Tek tissue culture Chamber/ Slides and Flaskette for processing cultured monolayers for electron microscopy

1983 ◽  
Vol 41 ◽  
pp. 626-627
Author(s):  
K. Chien ◽  
R.L. Van De Velde ◽  
R.C. Heusser ◽  
J.W. Said
Keyword(s):  
Tissue Culture ◽  
Glass Slide ◽  
Culture Chamber ◽  
Major Difficulty ◽  
Cell Monolayers ◽  
Plastic Resin ◽  
Fast Procedure ◽  
In Situ Embedding ◽  
Chamber Slide

In situ embedding is a preferable means for the ultrastructural study of cultured cell monolayers. However, the major difficulty is the separation of the polymerized plastic resin from the supporting substrates. This abstract demonstrates a simple, fast procedure for using existing products for cell culture and in situ embedding of monolayers. Epoxy block surfaces up to 10 cm2 can be easily separated from an untreated glass slide and sectioned vertically or horizontally for EM studies.The Lab-Tek tissue culture Chamber/Slides or Flaskette are constructed so that 1 to 8 chamber compartments or a single flask is attached to a regular glass slide by gaskets. Different cell lines or procedures can be performed on the same chamber/slide. When the plastic chambers and gasket are removed, the glass slide can be coverslipped and filed for reference. We use these products routinely for cell cultures. When an EM study is needed, we fix the cell monolayers in situ within the chambers or flask, dehydrate with ethanol, infiltrate and embed in an epoxy resin.

Microwave Processing of Cell Monolayers in Situ for Postembedding Immunocytochemistry with Retention of Ultrastructure and Antigenicity

1998 ◽  
Vol 4 (S2) ◽  
pp. 854-855
Author(s):  
Victoria J. Madden
Keyword(s):  
Electron Microscopy ◽  
Experimental Models ◽  
Microwave Energy ◽  
Microwave Oven ◽  
Biological Research ◽  
Biochemical Processes ◽  
Cell Monolayers ◽  
Acrylic Resins ◽  
Transmission Electron

Increasingly, cells isolated from blood and body fluids and cells grown in culture are becoming the experimental models of choice for biological research. The demand for demonstrating biochemical processes morphologically is also becoming commonplace in the electron microscopy laboratory. Successful fixation, in situ embedment, and ultrathinsectioning of cell monolayers can be difficult to achieve for routine transmission electron microscopy. For postembedding immunocytochemistry, processing becomes more complex due to fixation constraints and the use of acrylic resins. The object of this paper is to present a reliable, rapid method for processing monolayers that preserves both the ultrastructure of the cells and antigenicity.The equipment used for this procedure was a Pelco Model 3440 MAX laboratory microwave oven equipped with a temperature probe and a maximum power output of 800 watts. Using a neon bulb array, the oven cavity is calibrated to determine the microwave energy distribution.

A method for in situ embedding of cultured cells grown on plastic surfaces

In Vitro ◽  
1972 ◽  
Vol 8 (1) ◽  
pp. 26-29 ◽  
Author(s):  
William H. J. Douglas ◽  
John E. Elser
Keyword(s):  
Cultured Cells ◽  
Plastic Surfaces ◽  
In Situ Embedding

Immunocytochemistry. A Review of Methodology for Electron Microscopic Applicaiton

1974 ◽  
Vol 32 ◽  
pp. 328-329
Author(s):  
Joseph E. Mazurkiewicz
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Ultrastructural Level ◽  
Electron Microscopic ◽  
Biological Interest ◽  
Investigative Approach ◽  
Immunologic Activity ◽  
Dense Label

Immunocytochemistry is a powerful investigative approach in which one of the most exacting examples of specificity, that of the reaction of an antibody with its antigen, isused to localize tissue and cell specific molecules in situ. Following the introduction of fluorescent labeled antibodies in T950, a large number of molecules of biological interest had been studied with light microscopy, especially antigens involved in the pathogenesis of some diseases. However, with advances in electron microscopy, newer methods were needed which could reveal these reactions at the ultrastructural level. An electron dense label that could be coupled to an antibody without the loss of immunologic activity was desired.

Sign in / Sign up

Export Citation Format

Share Document