Optimization of glycol methacrylate embedding of large specimens in neurological research. Study of rat skull-brain specimens after implantation of polyester meshes.

ABSTRACT

Advances in neuroscience require better anatomical knowledge of neuronal architecture and structural details. Optimal embedding techniques are the basis for precise morphometric studies in section series as well as for the evaluation of tissue specimens or implants of differing hardness. There are very few methods for preparing large specimens by resin embedding, although resins such as polyethylene glycol (PEG) and methyl methacrylate (MMA) are presently in use. However, these methods have proven to be laborious and sometimes unsatisfactory for serial sectioning. While glycol methacrylate embedding (GMA) is suitable for smaller specimens, it results in inadequate infiltration and polymerization in blocks larger than 1 x 1 x 0.2 cm. We present an improved technique using GMA, which permits both standardized embedding of 4 x 2 x 2 cm blocks and preparation of section series. This method was developed for preserving skull-brain specimens from rats with polyester-mesh implants. The excellent preservation of cellular details allowed the assessment of local tissue reaction to foreign-body material in situ. Advantages of this method are (1) No toxic catalysts or solvents are used (as opposed to MMA and current GMA processes); (2) Laborious routines in stretching and mounting of sections are not necessary (in contrast to PEG and MMA); (3) No deplastination is required before staining (in contrast to PEG and MMA); (4) Excellent morphologic preservation of various tissue is achieved.