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Enhanced immunohistochemical resolution of claudin proteins in glycolmethacrylate-embedded tissue biopsies

Enhanced immunohistochemical resolution of claudin proteins in glycolmethacrylate-embedded tissue biopsies
Enhanced immunohistochemical resolution of claudin proteins in glycolmethacrylate-embedded tissue biopsies
There are a number of disadvantages with conventional tissue immunohistochemistry for accurate -localisation of claudin proteins. Traditionally, tissue cryopreservation or formaldehyde fixation with wax embedding is utilised prior to sectioning and antibody localisation. Wax embedding gives better morphological preservation than frozen tissue, but the required use of chemical cross-linking fixatives renders many antigens inaccessible to antibody binding or results in subsequent disruption of antibody localisation patterns due to the use of harsh antigen retrieval methods. Use of frozen or wax-embedded tissue also requires the cutting of relatively thick>6-?m sections, making the interrogation of serial sections very limited. The use of glycolmethacrylate (GMA) tissue embedding with fixation in acetone is compatible with epitope preservation for many antibody reagents that are often destroyed by chemical cross-linking fixatives. GMA is a water-miscible embedding resin that maintains tissue hydration during processing, thus reducing tissue shrinkage, while embedding and cutting in the polymerised resin physically supports the tissue, thus improving morphology. This method also facilitates the cutting of 2-?m sequential sections for analysis of multiple antigens and maximises the information available from small tissue biopsies from human clinical sources.
371-382
Humana
Collins, Jane E.
be0e66f1-3036-47fa-9d7e-914c48710ba4
Kirk, Adam
c9598ea9-e2dc-403f-8d37-8ea6407cb9cf
Campbell, Sara K.
5eb5a1ea-5833-40ed-8681-7af5c78a8c1f
Mason, Juan
3100af92-384e-44ff-ad79-02ba359b9bcf
Wilson, Susan J.
21c6875d-6870-441b-ae7a-603562a646b8
Turksen, K.
Collins, Jane E.
be0e66f1-3036-47fa-9d7e-914c48710ba4
Kirk, Adam
c9598ea9-e2dc-403f-8d37-8ea6407cb9cf
Campbell, Sara K.
5eb5a1ea-5833-40ed-8681-7af5c78a8c1f
Mason, Juan
3100af92-384e-44ff-ad79-02ba359b9bcf
Wilson, Susan J.
21c6875d-6870-441b-ae7a-603562a646b8
Turksen, K.

Collins, Jane E., Kirk, Adam, Campbell, Sara K., Mason, Juan and Wilson, Susan J. (2011) Enhanced immunohistochemical resolution of claudin proteins in glycolmethacrylate-embedded tissue biopsies. In, Turksen, K. (ed.) Claudins. Methods and Protocols. (Methods in Molecular Biology, 762) Totowa. Humana, pp. 371-382. (doi:10.1007/978-1-61779-185-7_27).

Record type: Book Section

Abstract

There are a number of disadvantages with conventional tissue immunohistochemistry for accurate -localisation of claudin proteins. Traditionally, tissue cryopreservation or formaldehyde fixation with wax embedding is utilised prior to sectioning and antibody localisation. Wax embedding gives better morphological preservation than frozen tissue, but the required use of chemical cross-linking fixatives renders many antigens inaccessible to antibody binding or results in subsequent disruption of antibody localisation patterns due to the use of harsh antigen retrieval methods. Use of frozen or wax-embedded tissue also requires the cutting of relatively thick>6-?m sections, making the interrogation of serial sections very limited. The use of glycolmethacrylate (GMA) tissue embedding with fixation in acetone is compatible with epitope preservation for many antibody reagents that are often destroyed by chemical cross-linking fixatives. GMA is a water-miscible embedding resin that maintains tissue hydration during processing, thus reducing tissue shrinkage, while embedding and cutting in the polymerised resin physically supports the tissue, thus improving morphology. This method also facilitates the cutting of 2-?m sequential sections for analysis of multiple antigens and maximises the information available from small tissue biopsies from human clinical sources.

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Published date: 2011
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 197325
URI: http://eprints.soton.ac.uk/id/eprint/197325
PURE UUID: 58429ddf-5594-4124-a4ba-33739b46bce9
ORCID for Susan J. Wilson: ORCID iD orcid.org/0000-0003-1305-8271

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Date deposited: 21 Sep 2011 14:36
Last modified: 15 Mar 2024 20:38

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Contributors

Author: Jane E. Collins
Author: Adam Kirk
Author: Sara K. Campbell
Author: Juan Mason
Author: Susan J. Wilson ORCID iD
Editor: K. Turksen

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