Use of Melinex film for flat embedding tissue sections in LR White.

LR White Melinex film acrylic resin flat embedding

Résumé

Tissue slices can undergo distortions during processing into resin for light and electron microscopy as a result of differential shrinkage of the various tissue components, and this may necessitate removal of a considerable amount of material from the final resin-embedded tissue block to ensure production of complete sections of the sample. To mitigate this problem, a number of techniques have been devised that ensure the sample is held flat during the final curing/polymerisation of the resin. For embedding in acrylic resins, oxygen must be excluded as it inhibits polymerisation, and methods devised for epoxy resin embedding are generally unsuitable. The method describes the preparation and use of air-tight flat-embedding chambers prepared from Melinex film and provides an inexpensive, technically simpler, and versatile alternative to chambers formed from either Thermanox coverslips or Aclar films that have previously been advocated for such purposes. Lay description: Tissue slices can undergo distortions during processing into resin for light and electron microscopy as a result of differential shrinkage of the various tissue components. Such distortions may necessitate removal of a considerable amount of material to ensure production of complete sections of the sample. For embedding in acrylic resins, oxygen must be excluded as it inhibits polymerisation, and methods devised for epoxy resin flat-embedding are generally unsuitable. Air-tight flat-embedding chambers prepared from either Thermanox coverslips, or a combination of PTFE-coated glass slides, polycarbonate film gaskets, and Aclar film have been advocated for such purposes. Thermanox coverslips are expensive and limited in size to 22 mm × 60 mm, and the alternative method is technically complicated. Melinex film is commercially available as 210 mm × 297 mm sheets and is approximately 1/20th the price of Thermanox and less than half the price of Aclar film. The method describes the preparation and use of embedding chambers made from Melinex film, glass slides and double-sided adhesive tape as a technically simpler, inexpensive and versatile alternative to both Thermanox coverslips and the Aclar film method.

Identifiants

DOI: 10.1111/jmi.13359 PMID: 39279683

pubmed: 39279683

doi: 10.1111/jmi.13359

doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Informations de copyright

Références

DeFelipe, J., & Fairén, A. (1993). A simple and reliable method for correlative light and electron microscopic studies. Journal of Histochemistry & Cytochemistry, 41(5), 769–772.

Holländer, H. (1970). The section embedding (SE) technique. A new method for the combined light microscopic and electron microscopic examination of central nervous tissue. Brain Research, 20(1), 39–47.

DeFelipe, J., Hendry, S. H., Jones, E. G., & Schmechel, D. (1985). Variability in the terminations of GABAergic chandelier cell axons on initial segments of pyramidal cell axons in the monkey sensory‐motor cortex. Journal of Comparative Neurology, 231, 364–384.

Mawe, G. M., Bresnahan, J. C., & Beattie, M. S. (1983). Ultrastructure of HRP‐Labelled Neurons: A comparison of two sensitive techniques. Brain Research Bulletin, 10(4), 551–558.

Aldes, L. D., & Boone, T. B. (1984). A combined flat‐embedding, HRP histochemical method for correlative light and electron microscopic study of single neurons. Journal of Neuroscience Research, 11(1), 27–34.

Sawada, H., & Esaki, M. (1998). A simple flat embedding method for the correlative light and electron microscopic immunocytochemistry. Journal of Electron Microscopy, 47, 535–537.

Newman, G. R., & Hobot, J. A. (2001). Resin microscopy and on‐section immunocytochemistry (2nd ed.). Springer‐Verlag.

Vesk, P. A., Dibbayawan, T. P., & Vesk, M. (1993). Flat embedding of microorganisms on slides with LR White for immunocytochemistry. Microscopy Research and Technique, 26(4), 352–353.

Palmieri, M., & Kiss, J. Z. (2005). A novel technique for flat‐embedding cryofixed plant specimens in LR White resin. Microscopy Research and Technique, 68(2), 80–84.

Firket, H. (1966). Polyester sheeting (Melinex O), a tissue‐culture support easily separable from epoxy resins after flat‐face embedding. Stain Technology, 41(3), 189–191.

Steiner, M., Schöfer, C., & Mosgoeller, W. (1994). In Situ Flat embedding of monolayers and cell relocation in the acrylic resin LR White for comparative light and electron microscopy studies. Histochemical Journal, 26, 934–938.

Use of Melinex film for flat embedding tissue sections in LR White.

Journal

Informations de publication