Methods for preparing standardized glycol methacrylate (GMA) based embedding media for use in light microscopy in a rational, precise and reproducible manner are described. The application of these procedures resulted in a versatile, low toxicity GMA embedding medium.
GMA embedding medium and resin blocks were tested utilizing a variety of physico-chemical techniques, namely: gas chromatography, determination of the maximum temperature reached during polymerization, the time taken to reach the maximum temperature, hardness testing, determination of the glass transition temperature, and measurement of the dimensional changes following section stretching and mounting at various temperatures. Data obtained from these techniques enabled a multi-purpose GMA embedding medium to be precisely specified.
The infiltration solution, as well as the accelerator solution, of this mixture contains fewer toxic components than comparable systems. The infiltration solution consists of GMA monomer and a non-toxic plasticizer (2-isopropoxy ethanol). The initiator is a 50% damped dibenzoyl peroxide, and the accelerator solution is composed of a mixture of polyethylene glycol 200 and a low concentration of a low toxicity tertiary amine (N ,N ,3,5-tetramethylaniline). Resin blocks obtained from this mixture are highly transparent, do not become coloured, and are easy to cut at 0.5-2.0 μm.
Various histological techniques, such as routine embedding of implanted biomaterials, histochemistry, enzyme histochemistry, and immunohistochemistry were carried out; the new GMA embedding medium proved to be applicable in all the techniques without laborious modifications.
Gerrits, Peter O.; Eppinger, Bernhard; Goor, Harry van; and Horobin, Richard W.
"A Versatile, Low Toxicity Glycol Methacrylate Embedding Medium for Use in Biological Research, and for Recovered Biomaterials Prostheses,"
Cells and Materials: Vol. 1
, Article 1.
Available at: http://digitalcommons.usu.edu/cellsandmaterials/vol1/iss3/1