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Butyl-methyl-methacrylate for Immunocytochemistry Through the Light Microscope

Published online by Cambridge University Press:  14 March 2018

Tobias I. Baskin*
Affiliation:
University of Massachusetts, Amherst, MA

Extract

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The use of methacrylate monomers for embedding has a venerable history in microscopy. Many formulations have been developed over the years for various purposes, ranging from standard TEM observations to low-temperature embedding. Key parameters include the length of the hydrocarbon chain and the presence and kind of cross linking reagent. In the mixture of butyl and methyl methacrylate (BMM) described here, the monomers are relatively short-chained and there is no cross linker at all. This gives the polymerized material a softness that makes it rather unsuitable for TEM, but on the contrary allows the embedment to be removed after sectioning by a brief incubation in acetone. The latter property is good for immunocytochemistry because loss of the embedment means greater access for the antibody to the antigen. BMM generally preserves structure better than paraffin or glycol-methacrylate, and for this reason is a useful choice for light-level immunocytochemistry, particularly when sub-cellular resolution is desired.

Type
Microscopy 101
Copyright
Copyright © Microscopy Society of America 2006

References

Citations

Baskin, TI, Busby, CH, Fowke, LC, Sammut, M, Gubler, F, (1992) Improvements in immunostaining samples embedded in methacrylate: Localization of microtubules and other antigens throughout developing organs in plants of diverse taxa. Planta, Vol 187 405413.Google Scholar
Baskin, TI, Miller, DD, Vos, JW, Wilson, JE, Hepler, PK, (1996) Cryofixing single cells and multicellular specimens enhances structure & immunocytochemistry for light microscopy. J. Microsc., Vol 182 149161.Google Scholar
Kronenberger, J, Desprez, T, Hofte, H, Caboche, M, Traas, J, (1993) A methacrylate embedding procedure developed for immunolocalization on plant-tissues is also compatible with in-situ hybridization. Cell Biol. Internat., Vol 17 10131021.CrossRefGoogle Scholar
Palmer, JH, Harper, JDI, Marc, J, (2001) Control of brittleness in butyl-methyl-methacrylate resin embedding mixtures to facilitate their use in immunofluorescence microscopy. Cytobios, Vol 104 145156.Google Scholar