Microwave antigen retrieval in immunocytochemistry: a study of 80 antibodies
AIMS--To evaluate the effect of microwave irradiation on the staining quality of a range of commonly used primary antibodies in archival, formalin fixed, paraffin wax embedded material, with emphasis on antibodies that have previously worked successfully only on frozen tissue. METHODS--Immunocytoche...
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| Veröffentlicht in: | Journal of clinical pathology 1994-05, Vol.47 (5), p.448-452 |
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| container_title | Journal of clinical pathology |
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| creator | Cuevas, E C Bateman, A C Wilkins, B S Johnson, P A Williams, J H Lee, A H Jones, D B Wright, D H |
| description | AIMS--To evaluate the effect of microwave irradiation on the staining quality of a range of commonly used primary antibodies in archival, formalin fixed, paraffin wax embedded material, with emphasis on antibodies that have previously worked successfully only on frozen tissue. METHODS--Immunocytochemistry (streptavidin-biotin complex technique) was performed on histological sections of a range of normal and pathological tissues, after varying treatment with microwave irradiation. The staining quality of each antibody was compared with that achieved without prior treatment of the sections or after enzyme predigestion. RESULTS--Microwave irradiation permitted successful immunostaining with 20 antibodies that stained only frozen tissues before. The staining characteristics of 21 antibodies that were already known to stain formalin fixed, paraffin wax embedded material were improved. Another 39 antibodies did not show enhanced staining with microwave irradiation. The method preserves tissue morphology and produces more consistent staining than that achieved by enzyme predigestion with many antibodies. Microwave irradiation may also allow some primary antibodies to be used at higher working dilutions. The citrate buffer used in this study avoids the necessity of exposure to heavy metal salts. CONCLUSIONS--Microwave antigen retrieval represents an important technical advance within immunocytochemistry that will greatly increase the range of antibodies which can be used to study formalin fixed, paraffin wax embedded tissues. |
| doi_str_mv | 10.1136/jcp.47.5.448 |
| format | Article |
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METHODS--Immunocytochemistry (streptavidin-biotin complex technique) was performed on histological sections of a range of normal and pathological tissues, after varying treatment with microwave irradiation. The staining quality of each antibody was compared with that achieved without prior treatment of the sections or after enzyme predigestion. RESULTS--Microwave irradiation permitted successful immunostaining with 20 antibodies that stained only frozen tissues before. The staining characteristics of 21 antibodies that were already known to stain formalin fixed, paraffin wax embedded material were improved. Another 39 antibodies did not show enhanced staining with microwave irradiation. The method preserves tissue morphology and produces more consistent staining than that achieved by enzyme predigestion with many antibodies. Microwave irradiation may also allow some primary antibodies to be used at higher working dilutions. The citrate buffer used in this study avoids the necessity of exposure to heavy metal salts. CONCLUSIONS--Microwave antigen retrieval represents an important technical advance within immunocytochemistry that will greatly increase the range of antibodies which can be used to study formalin fixed, paraffin wax embedded tissues.</description><identifier>ISSN: 0021-9746</identifier><identifier>EISSN: 1472-4146</identifier><identifier>DOI: 10.1136/jcp.47.5.448</identifier><identifier>PMID: 7517960</identifier><identifier>CODEN: JCPAAK</identifier><language>eng</language><publisher>London: BMJ Publishing Group Ltd and Association of Clinical Pathologists</publisher><subject>Antibodies, Monoclonal ; Antigens - analysis ; Antigens - radiation effects ; Antigens, Neoplasm - analysis ; Biological and medical sciences ; Formaldehyde ; Humans ; Immunohistochemical methods ; Immunohistochemistry - methods ; Immunological methods for diagnosis and exploration ; Immunopathology ; Lymphoma - immunology ; Medical sciences ; Microwaves ; Palatine Tonsil - immunology ; Paraffin Embedding ; Staining and Labeling - methods ; Thyroid Gland - immunology</subject><ispartof>Journal of clinical pathology, 1994-05, Vol.47 (5), p.448-452</ispartof><rights>1995 INIST-CNRS</rights><rights>Copyright BMJ Publishing Group LTD May 1994</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-b505t-ccec30a9e60601c68509fdc77e9be746d37fca17ccff647fb3e888cf584439ab3</citedby><cites>FETCH-LOGICAL-b505t-ccec30a9e60601c68509fdc77e9be746d37fca17ccff647fb3e888cf584439ab3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC502024/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC502024/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=3386776$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7517960$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Cuevas, E C</creatorcontrib><creatorcontrib>Bateman, A C</creatorcontrib><creatorcontrib>Wilkins, B S</creatorcontrib><creatorcontrib>Johnson, P A</creatorcontrib><creatorcontrib>Williams, J H</creatorcontrib><creatorcontrib>Lee, A H</creatorcontrib><creatorcontrib>Jones, D B</creatorcontrib><creatorcontrib>Wright, D H</creatorcontrib><title>Microwave antigen retrieval in immunocytochemistry: a study of 80 antibodies</title><title>Journal of clinical pathology</title><addtitle>J Clin Pathol</addtitle><description>AIMS--To evaluate the effect of microwave irradiation on the staining quality of a range of commonly used primary antibodies in archival, formalin fixed, paraffin wax embedded material, with emphasis on antibodies that have previously worked successfully only on frozen tissue. METHODS--Immunocytochemistry (streptavidin-biotin complex technique) was performed on histological sections of a range of normal and pathological tissues, after varying treatment with microwave irradiation. The staining quality of each antibody was compared with that achieved without prior treatment of the sections or after enzyme predigestion. RESULTS--Microwave irradiation permitted successful immunostaining with 20 antibodies that stained only frozen tissues before. The staining characteristics of 21 antibodies that were already known to stain formalin fixed, paraffin wax embedded material were improved. Another 39 antibodies did not show enhanced staining with microwave irradiation. The method preserves tissue morphology and produces more consistent staining than that achieved by enzyme predigestion with many antibodies. Microwave irradiation may also allow some primary antibodies to be used at higher working dilutions. The citrate buffer used in this study avoids the necessity of exposure to heavy metal salts. CONCLUSIONS--Microwave antigen retrieval represents an important technical advance within immunocytochemistry that will greatly increase the range of antibodies which can be used to study formalin fixed, paraffin wax embedded tissues.</description><subject>Antibodies, Monoclonal</subject><subject>Antigens - analysis</subject><subject>Antigens - radiation effects</subject><subject>Antigens, Neoplasm - analysis</subject><subject>Biological and medical sciences</subject><subject>Formaldehyde</subject><subject>Humans</subject><subject>Immunohistochemical methods</subject><subject>Immunohistochemistry - methods</subject><subject>Immunological methods for diagnosis and exploration</subject><subject>Immunopathology</subject><subject>Lymphoma - immunology</subject><subject>Medical sciences</subject><subject>Microwaves</subject><subject>Palatine Tonsil - immunology</subject><subject>Paraffin Embedding</subject><subject>Staining and Labeling - methods</subject><subject>Thyroid Gland - immunology</subject><issn>0021-9746</issn><issn>1472-4146</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kc1v1DAQxS1EVbaFG1ekSKByIYsdf4yDxAEWCkgLCAkQ4mI5jt16SeLFTrbsf4_bXa2AA6c5vN8bvZmH0H2C54RQ8XRl1nMGcz5nTN5CM8KgKhlh4jaaYVyRsgYm7qCTlFYYEwqEHqNj4ARqgWdo-d6bGK70xhZ6GP2FHYpox-jtRneFHwrf99MQzHYM5tL2Po1x-6zQRRqndlsEV0h842tC6226i46c7pK9t5-n6Mv568-Lt-Xy45t3ixfLsuGYj6Ux1lCsayuwwMQIyXHtWgNg68bmsC0FZzQBY5wTDFxDrZTSOC4Zo7Vu6Cl6vtu7npretsYOY9SdWkff67hVQXv1tzL4S3URNorjClcs-8_2_hh-TjaNKl9mbNfpwYYpKRBc1qSGDD78B1yFKQ75NkUAMOO0ptfrnuyo_MmUonWHJASr64pUrkgxUFzlijL-4M_0B3jfSdYf7XWdjO5c1IPx6YBRKgWAyFi5w3Ip9tdB1vGHEkCBqw9fFwpevpLVp-9Efcv84x3f9Kv_B_wNdDy2og</recordid><startdate>19940501</startdate><enddate>19940501</enddate><creator>Cuevas, E C</creator><creator>Bateman, A C</creator><creator>Wilkins, B S</creator><creator>Johnson, P A</creator><creator>Williams, J H</creator><creator>Lee, A H</creator><creator>Jones, D B</creator><creator>Wright, D H</creator><general>BMJ Publishing Group Ltd and Association of Clinical Pathologists</general><general>BMJ</general><general>BMJ Publishing Group LTD</general><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>88I</scope><scope>8AF</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>BTHHO</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19940501</creationdate><title>Microwave antigen retrieval in immunocytochemistry: a study of 80 antibodies</title><author>Cuevas, E C ; Bateman, A C ; Wilkins, B S ; Johnson, P A ; Williams, J H ; Lee, A H ; Jones, D B ; Wright, D H</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-b505t-ccec30a9e60601c68509fdc77e9be746d37fca17ccff647fb3e888cf584439ab3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>Antibodies, Monoclonal</topic><topic>Antigens - analysis</topic><topic>Antigens - radiation effects</topic><topic>Antigens, Neoplasm - analysis</topic><topic>Biological and medical sciences</topic><topic>Formaldehyde</topic><topic>Humans</topic><topic>Immunohistochemical methods</topic><topic>Immunohistochemistry - methods</topic><topic>Immunological methods for diagnosis and exploration</topic><topic>Immunopathology</topic><topic>Lymphoma - immunology</topic><topic>Medical sciences</topic><topic>Microwaves</topic><topic>Palatine Tonsil - immunology</topic><topic>Paraffin Embedding</topic><topic>Staining and Labeling - methods</topic><topic>Thyroid Gland - immunology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Cuevas, E C</creatorcontrib><creatorcontrib>Bateman, A C</creatorcontrib><creatorcontrib>Wilkins, B S</creatorcontrib><creatorcontrib>Johnson, P A</creatorcontrib><creatorcontrib>Williams, J H</creatorcontrib><creatorcontrib>Lee, A H</creatorcontrib><creatorcontrib>Jones, D B</creatorcontrib><creatorcontrib>Wright, D H</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>STEM Database</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>ProQuest Central</collection><collection>BMJ Journals</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of clinical pathology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Cuevas, E C</au><au>Bateman, A C</au><au>Wilkins, B S</au><au>Johnson, P A</au><au>Williams, J H</au><au>Lee, A H</au><au>Jones, D B</au><au>Wright, D H</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Microwave antigen retrieval in immunocytochemistry: a study of 80 antibodies</atitle><jtitle>Journal of clinical pathology</jtitle><addtitle>J Clin Pathol</addtitle><date>1994-05-01</date><risdate>1994</risdate><volume>47</volume><issue>5</issue><spage>448</spage><epage>452</epage><pages>448-452</pages><issn>0021-9746</issn><eissn>1472-4146</eissn><coden>JCPAAK</coden><abstract>AIMS--To evaluate the effect of microwave irradiation on the staining quality of a range of commonly used primary antibodies in archival, formalin fixed, paraffin wax embedded material, with emphasis on antibodies that have previously worked successfully only on frozen tissue. METHODS--Immunocytochemistry (streptavidin-biotin complex technique) was performed on histological sections of a range of normal and pathological tissues, after varying treatment with microwave irradiation. The staining quality of each antibody was compared with that achieved without prior treatment of the sections or after enzyme predigestion. RESULTS--Microwave irradiation permitted successful immunostaining with 20 antibodies that stained only frozen tissues before. The staining characteristics of 21 antibodies that were already known to stain formalin fixed, paraffin wax embedded material were improved. Another 39 antibodies did not show enhanced staining with microwave irradiation. The method preserves tissue morphology and produces more consistent staining than that achieved by enzyme predigestion with many antibodies. Microwave irradiation may also allow some primary antibodies to be used at higher working dilutions. The citrate buffer used in this study avoids the necessity of exposure to heavy metal salts. CONCLUSIONS--Microwave antigen retrieval represents an important technical advance within immunocytochemistry that will greatly increase the range of antibodies which can be used to study formalin fixed, paraffin wax embedded tissues.</abstract><cop>London</cop><pub>BMJ Publishing Group Ltd and Association of Clinical Pathologists</pub><pmid>7517960</pmid><doi>10.1136/jcp.47.5.448</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Journal of clinical pathology, 1994-05, Vol.47 (5), p.448-452 |
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| subjects | Antibodies, Monoclonal Antigens - analysis Antigens - radiation effects Antigens, Neoplasm - analysis Biological and medical sciences Formaldehyde Humans Immunohistochemical methods Immunohistochemistry - methods Immunological methods for diagnosis and exploration Immunopathology Lymphoma - immunology Medical sciences Microwaves Palatine Tonsil - immunology Paraffin Embedding Staining and Labeling - methods Thyroid Gland - immunology |
| title | Microwave antigen retrieval in immunocytochemistry: a study of 80 antibodies |
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