A simplified technique for histologic analysis of central nervous system tissues using glycol-methacrylate plastic coupled with pre-embedding immunocytochemistry
Paraffin and some plastic embedding techniques will destroy many antigens routinely detected by immunocytochemistry performed on frozen tissue sections. However, morphologic quality is compromised to varying extents in frozen tissue, even with the use of cryoprotection. We report a simple glycol-met...
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| Veröffentlicht in: | Journal of neuroscience methods 1994-09, Vol.54 (1), p.23-29 |
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| creator | Abbott, Louise C Conforti, Michael L Isaacs, Krystyna R Crawley, Jacqueline N Sterchi, Diane |
| description | Paraffin and some plastic embedding techniques will destroy many antigens routinely detected by immunocytochemistry performed on frozen tissue sections. However, morphologic quality is compromised to varying extents in frozen tissue, even with the use of cryoprotection. We report a simple glycol-methacrylate (GMA) embedding technique using vibratome-sectioned mouse brain reacted for tyrosine hydroxylase (TH) immunoreactivity before plastic embedding. In this study we used a short (4 h) simple, GMA embedding procedure which subsequently provided 1.5–5.0 μm sections yielding morphologic details superior to frozen or paraffin sections. Prior to embedding we used a peroxidase-antiperoxidase (PAP) reaction with the 3,3′ diaminobenzidine tetrahydrochloride (DAB) chromogen visualizing TH. Several different counterstains were used, demonstrating the versatility of this embedding procedure. |
| doi_str_mv | 10.1016/0165-0270(94)90156-2 |
| format | Article |
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However, morphologic quality is compromised to varying extents in frozen tissue, even with the use of cryoprotection. We report a simple glycol-methacrylate (GMA) embedding technique using vibratome-sectioned mouse brain reacted for tyrosine hydroxylase (TH) immunoreactivity before plastic embedding. In this study we used a short (4 h) simple, GMA embedding procedure which subsequently provided 1.5–5.0 μm sections yielding morphologic details superior to frozen or paraffin sections. Prior to embedding we used a peroxidase-antiperoxidase (PAP) reaction with the 3,3′ diaminobenzidine tetrahydrochloride (DAB) chromogen visualizing TH. 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Methods ; Glycol methacrylate ; Histological Techniques ; Immunoenzyme Techniques ; Male ; Methacrylate ; Methacrylates ; Mice ; Mice, Inbred C57BL ; Peroxidase-antiperoxidase ; Plastic Embedding ; Staining and Labeling ; Tyrosine 3-Monooxygenase - metabolism ; Tyrosine hydroxylase ; Vertebrates: nervous system and sense organs</subject><ispartof>Journal of neuroscience methods, 1994-09, Vol.54 (1), p.23-29</ispartof><rights>1994</rights><rights>1995 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c301t-f9afc8b8e769eb1ee9f1bbed11b317e18fd51bb60f0426896ca860d231c0cf553</citedby><cites>FETCH-LOGICAL-c301t-f9afc8b8e769eb1ee9f1bbed11b317e18fd51bb60f0426896ca860d231c0cf553</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/0165027094901562$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=3321116$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7529349$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Abbott, Louise C</creatorcontrib><creatorcontrib>Conforti, Michael L</creatorcontrib><creatorcontrib>Isaacs, Krystyna R</creatorcontrib><creatorcontrib>Crawley, Jacqueline N</creatorcontrib><creatorcontrib>Sterchi, Diane</creatorcontrib><title>A simplified technique for histologic analysis of central nervous system tissues using glycol-methacrylate plastic coupled with pre-embedding immunocytochemistry</title><title>Journal of neuroscience methods</title><addtitle>J Neurosci Methods</addtitle><description>Paraffin and some plastic embedding techniques will destroy many antigens routinely detected by immunocytochemistry performed on frozen tissue sections. However, morphologic quality is compromised to varying extents in frozen tissue, even with the use of cryoprotection. We report a simple glycol-methacrylate (GMA) embedding technique using vibratome-sectioned mouse brain reacted for tyrosine hydroxylase (TH) immunoreactivity before plastic embedding. In this study we used a short (4 h) simple, GMA embedding procedure which subsequently provided 1.5–5.0 μm sections yielding morphologic details superior to frozen or paraffin sections. Prior to embedding we used a peroxidase-antiperoxidase (PAP) reaction with the 3,3′ diaminobenzidine tetrahydrochloride (DAB) chromogen visualizing TH. Several different counterstains were used, demonstrating the versatility of this embedding procedure.</description><subject>3,3'-Diaminobenzidine</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Brain</subject><subject>Brain - enzymology</subject><subject>Brain - pathology</subject><subject>Catecholamine neuron</subject><subject>Cerebellum</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>General aspects. Models. Methods</subject><subject>Glycol methacrylate</subject><subject>Histological Techniques</subject><subject>Immunoenzyme Techniques</subject><subject>Male</subject><subject>Methacrylate</subject><subject>Methacrylates</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Peroxidase-antiperoxidase</subject><subject>Plastic Embedding</subject><subject>Staining and Labeling</subject><subject>Tyrosine 3-Monooxygenase - metabolism</subject><subject>Tyrosine hydroxylase</subject><subject>Vertebrates: nervous system and sense organs</subject><issn>0165-0270</issn><issn>1872-678X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9UcuKFDEUDaKMPaN_oJCFyLgoTeqRqmwGhsEXDLhRcBdSqZuuSKpS5qZG6nP8U9PTTS9dhEDuedycQ8grzt5zxsWHfJqClS27lvU7yXgjivIJ2fGuLQvRdj-fkt0Z8pxcIv5ijNWSiQty0TalrGq5I39vKbpp8c46GGgCM87u9wrUhkhHhyn4sHeG6ln7DR3SYKmBOUXt6QzxIaxIccMEE00OcQWkK7p5T_d-M8EXE6RRm7h5nYAuXmPKYiasi89uf1wa6RKhgKmHYTjQ3DStczBbCmaEKfvH7QV5ZrVHeHm6r8iPTx-_330p7r99_np3e1-YivFUWKmt6foOWiGh5wDS8j7Lct5XvAXe2aHJD4JZVpeik8LoTrChrLhhxjZNdUXeHnWXGHICmFT2N-C9niF_U2XdipePwPoINDEgRrBqiW7ScVOcqUMz6hC7OsSuZK0em1Flpr0-6a_9BMOZdKoiz9-c5hqN9jbq2Tg8w6qq5JyLDLs5wiBn8eAgKjQOZgODi2CSGoL7_x7_AJc4sCI</recordid><startdate>199409</startdate><enddate>199409</enddate><creator>Abbott, Louise C</creator><creator>Conforti, Michael L</creator><creator>Isaacs, Krystyna R</creator><creator>Crawley, Jacqueline N</creator><creator>Sterchi, Diane</creator><general>Elsevier B.V</general><general>Elsevier Science</general><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>7X8</scope></search><sort><creationdate>199409</creationdate><title>A simplified technique for histologic analysis of central nervous system tissues using glycol-methacrylate plastic coupled with pre-embedding immunocytochemistry</title><author>Abbott, Louise C ; Conforti, Michael L ; Isaacs, Krystyna R ; Crawley, Jacqueline N ; Sterchi, Diane</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c301t-f9afc8b8e769eb1ee9f1bbed11b317e18fd51bb60f0426896ca860d231c0cf553</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>3,3'-Diaminobenzidine</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Brain</topic><topic>Brain - enzymology</topic><topic>Brain - pathology</topic><topic>Catecholamine neuron</topic><topic>Cerebellum</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>General aspects. Models. Methods</topic><topic>Glycol methacrylate</topic><topic>Histological Techniques</topic><topic>Immunoenzyme Techniques</topic><topic>Male</topic><topic>Methacrylate</topic><topic>Methacrylates</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Peroxidase-antiperoxidase</topic><topic>Plastic Embedding</topic><topic>Staining and Labeling</topic><topic>Tyrosine 3-Monooxygenase - metabolism</topic><topic>Tyrosine hydroxylase</topic><topic>Vertebrates: nervous system and sense organs</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Abbott, Louise C</creatorcontrib><creatorcontrib>Conforti, Michael L</creatorcontrib><creatorcontrib>Isaacs, Krystyna R</creatorcontrib><creatorcontrib>Crawley, Jacqueline N</creatorcontrib><creatorcontrib>Sterchi, Diane</creatorcontrib><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>MEDLINE - Academic</collection><jtitle>Journal of neuroscience methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Abbott, Louise C</au><au>Conforti, Michael L</au><au>Isaacs, Krystyna R</au><au>Crawley, Jacqueline N</au><au>Sterchi, Diane</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A simplified technique for histologic analysis of central nervous system tissues using glycol-methacrylate plastic coupled with pre-embedding immunocytochemistry</atitle><jtitle>Journal of neuroscience methods</jtitle><addtitle>J Neurosci Methods</addtitle><date>1994-09</date><risdate>1994</risdate><volume>54</volume><issue>1</issue><spage>23</spage><epage>29</epage><pages>23-29</pages><issn>0165-0270</issn><eissn>1872-678X</eissn><coden>JNMEDT</coden><abstract>Paraffin and some plastic embedding techniques will destroy many antigens routinely detected by immunocytochemistry performed on frozen tissue sections. However, morphologic quality is compromised to varying extents in frozen tissue, even with the use of cryoprotection. We report a simple glycol-methacrylate (GMA) embedding technique using vibratome-sectioned mouse brain reacted for tyrosine hydroxylase (TH) immunoreactivity before plastic embedding. In this study we used a short (4 h) simple, GMA embedding procedure which subsequently provided 1.5–5.0 μm sections yielding morphologic details superior to frozen or paraffin sections. Prior to embedding we used a peroxidase-antiperoxidase (PAP) reaction with the 3,3′ diaminobenzidine tetrahydrochloride (DAB) chromogen visualizing TH. Several different counterstains were used, demonstrating the versatility of this embedding procedure.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>7529349</pmid><doi>10.1016/0165-0270(94)90156-2</doi><tpages>7</tpages></addata></record> |
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| subjects | 3,3'-Diaminobenzidine Animals Biological and medical sciences Brain Brain - enzymology Brain - pathology Catecholamine neuron Cerebellum Fundamental and applied biological sciences. Psychology General aspects. Models. Methods Glycol methacrylate Histological Techniques Immunoenzyme Techniques Male Methacrylate Methacrylates Mice Mice, Inbred C57BL Peroxidase-antiperoxidase Plastic Embedding Staining and Labeling Tyrosine 3-Monooxygenase - metabolism Tyrosine hydroxylase Vertebrates: nervous system and sense organs |
| title | A simplified technique for histologic analysis of central nervous system tissues using glycol-methacrylate plastic coupled with pre-embedding immunocytochemistry |
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