Alterations of mouse lung tissue dimensions during processing for morphometry: a comparison of methods
Preservation of original tissue dimensions is an essential prerequisite for morphometric studies. Shrinkage occurring during tissue processing for histology may severely influence the appearance of structures seen under the microscope and stereological calculations. Therefore, shrinkage has to be av...
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| Veröffentlicht in: | American journal of physiology. Lung cellular and molecular physiology 2014-02, Vol.306 (4), p.L341-L350 |
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| container_end_page | L350 |
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| container_title | American journal of physiology. Lung cellular and molecular physiology |
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| creator | Schneider, Jan Philipp Ochs, Matthias |
| description | Preservation of original tissue dimensions is an essential prerequisite for morphometric studies. Shrinkage occurring during tissue processing for histology may severely influence the appearance of structures seen under the microscope and stereological calculations. Therefore, shrinkage has to be avoided so that estimates obtained by application of unbiased stereology are indeed unbiased. The present study investigates the alterations of tissue dimensions of mouse lung samples during processing for histology. Different fixatives as well as embedding protocols are considered. Mouse lungs were fixed by instillation of either 4% formalin or a mixture of 1.5% glutaraldehyde/1.5% formaldehyde. Tissue blocks were sampled according to principles of stereology for embedding in paraffin, glycol methacrylate without treatment with osmium tetroxide and uranyl acetate, and glycol methacrylate including treatment with osmium tetroxide and uranyl acetate before dehydration. Shrinkage was investigated by stereological measurements of dimensional changes of tissue cut faces. Results show a shrinkage of the cut face areas of roughly 40% per lung during paraffin embedding, 30% during "simple" glycol methacrylate embedding, and |
| doi_str_mv | 10.1152/ajplung.00329.2013 |
| format | Article |
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Shrinkage occurring during tissue processing for histology may severely influence the appearance of structures seen under the microscope and stereological calculations. Therefore, shrinkage has to be avoided so that estimates obtained by application of unbiased stereology are indeed unbiased. The present study investigates the alterations of tissue dimensions of mouse lung samples during processing for histology. Different fixatives as well as embedding protocols are considered. Mouse lungs were fixed by instillation of either 4% formalin or a mixture of 1.5% glutaraldehyde/1.5% formaldehyde. Tissue blocks were sampled according to principles of stereology for embedding in paraffin, glycol methacrylate without treatment with osmium tetroxide and uranyl acetate, and glycol methacrylate including treatment with osmium tetroxide and uranyl acetate before dehydration. Shrinkage was investigated by stereological measurements of dimensional changes of tissue cut faces. Results show a shrinkage of the cut face areas of roughly 40% per lung during paraffin embedding, 30% during "simple" glycol methacrylate embedding, and <3% during osmium tetroxide/uranyl acetate/glycol methacrylate embedding. Furthermore, the superiority of the glutaraldehyde-containing fixative regarding shrinkage is demonstrated. In conclusion, the use of a glutaraldehyde-containing fixative and embedding in glycol methacrylate with previous treatment of the samples with osmium tetroxide and uranyl acetate before dehydration is recommended for stereological studies of the mouse lung.</description><identifier>ISSN: 1040-0605</identifier><identifier>EISSN: 1522-1504</identifier><identifier>DOI: 10.1152/ajplung.00329.2013</identifier><identifier>PMID: 24375800</identifier><language>eng</language><publisher>United States: American Physiological Society</publisher><subject>Animals ; Comparative analysis ; Desiccation ; Fixatives - chemistry ; Glutaral - chemistry ; Histology ; Lung - anatomy & histology ; Lungs ; Male ; Methacrylates - chemistry ; Mice ; Mice, Inbred C57BL ; Organ Size ; Organometallic Compounds - chemistry ; Osmium Tetroxide - chemistry ; Paraffin Embedding ; Physiology ; Plastic Embedding ; Rodents ; Specimen Handling ; Tissue Fixation ; Tissues</subject><ispartof>American journal of physiology. Lung cellular and molecular physiology, 2014-02, Vol.306 (4), p.L341-L350</ispartof><rights>Copyright American Physiological Society Feb 15, 2014</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c397t-89aa5a12682c9bc258c802a2d8d6060258d141bcd08e7103ee18232ec76830873</citedby><cites>FETCH-LOGICAL-c397t-89aa5a12682c9bc258c802a2d8d6060258d141bcd08e7103ee18232ec76830873</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,3037,27923,27924</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24375800$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Schneider, Jan Philipp</creatorcontrib><creatorcontrib>Ochs, Matthias</creatorcontrib><title>Alterations of mouse lung tissue dimensions during processing for morphometry: a comparison of methods</title><title>American journal of physiology. Lung cellular and molecular physiology</title><addtitle>Am J Physiol Lung Cell Mol Physiol</addtitle><description>Preservation of original tissue dimensions is an essential prerequisite for morphometric studies. Shrinkage occurring during tissue processing for histology may severely influence the appearance of structures seen under the microscope and stereological calculations. Therefore, shrinkage has to be avoided so that estimates obtained by application of unbiased stereology are indeed unbiased. The present study investigates the alterations of tissue dimensions of mouse lung samples during processing for histology. Different fixatives as well as embedding protocols are considered. Mouse lungs were fixed by instillation of either 4% formalin or a mixture of 1.5% glutaraldehyde/1.5% formaldehyde. Tissue blocks were sampled according to principles of stereology for embedding in paraffin, glycol methacrylate without treatment with osmium tetroxide and uranyl acetate, and glycol methacrylate including treatment with osmium tetroxide and uranyl acetate before dehydration. Shrinkage was investigated by stereological measurements of dimensional changes of tissue cut faces. Results show a shrinkage of the cut face areas of roughly 40% per lung during paraffin embedding, 30% during "simple" glycol methacrylate embedding, and <3% during osmium tetroxide/uranyl acetate/glycol methacrylate embedding. Furthermore, the superiority of the glutaraldehyde-containing fixative regarding shrinkage is demonstrated. In conclusion, the use of a glutaraldehyde-containing fixative and embedding in glycol methacrylate with previous treatment of the samples with osmium tetroxide and uranyl acetate before dehydration is recommended for stereological studies of the mouse lung.</description><subject>Animals</subject><subject>Comparative analysis</subject><subject>Desiccation</subject><subject>Fixatives - chemistry</subject><subject>Glutaral - chemistry</subject><subject>Histology</subject><subject>Lung - anatomy & histology</subject><subject>Lungs</subject><subject>Male</subject><subject>Methacrylates - chemistry</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Organ Size</subject><subject>Organometallic Compounds - chemistry</subject><subject>Osmium Tetroxide - chemistry</subject><subject>Paraffin Embedding</subject><subject>Physiology</subject><subject>Plastic Embedding</subject><subject>Rodents</subject><subject>Specimen Handling</subject><subject>Tissue Fixation</subject><subject>Tissues</subject><issn>1040-0605</issn><issn>1522-1504</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdUTtPwzAQthCIlsIfYECRWFhSznac2Gyo4iVVYoHZcp0LTZXEwU6G_nvcBwxMd6fvobv7CLmmMKdUsHuz6Zux-5oDcKbmDCg_IdMIsJQKyE5jDxmkkIOYkIsQNgAgAPJzMmEZL4QEmJLqsRnQm6F2XUhclbRuDJjsbJOhDmHEpKxb7MIeL0dfR6D3zmIIu7ZyPkp8v3YtDn77kJjEurY3vg6u2_vhsHZluCRnlWkCXh3rjHw-P30sXtPl-8vb4nGZWq6KIZXKGGEoyyWzamWZkFYCM6yUZR7viHNJM7qyJUgsKHBEKhlnaItccpAFn5G7g2_c8XvEMOi2DhabxnQYL9M0U4oKBZRG6u0_6saNvovb6fg-UAK4gshiB5b1LgSPle593Rq_1RT0LgV9TEHvU9C7FKLo5mg9rlos_yS_b-c_CVSEuw</recordid><startdate>20140215</startdate><enddate>20140215</enddate><creator>Schneider, Jan Philipp</creator><creator>Ochs, Matthias</creator><general>American Physiological Society</general><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>7QP</scope><scope>7TS</scope><scope>7U7</scope><scope>C1K</scope><scope>7X8</scope></search><sort><creationdate>20140215</creationdate><title>Alterations of mouse lung tissue dimensions during processing for morphometry: a comparison of methods</title><author>Schneider, Jan Philipp ; Ochs, Matthias</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c397t-89aa5a12682c9bc258c802a2d8d6060258d141bcd08e7103ee18232ec76830873</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Animals</topic><topic>Comparative analysis</topic><topic>Desiccation</topic><topic>Fixatives - chemistry</topic><topic>Glutaral - chemistry</topic><topic>Histology</topic><topic>Lung - anatomy & histology</topic><topic>Lungs</topic><topic>Male</topic><topic>Methacrylates - chemistry</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Organ Size</topic><topic>Organometallic Compounds - chemistry</topic><topic>Osmium Tetroxide - chemistry</topic><topic>Paraffin Embedding</topic><topic>Physiology</topic><topic>Plastic Embedding</topic><topic>Rodents</topic><topic>Specimen Handling</topic><topic>Tissue Fixation</topic><topic>Tissues</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Schneider, Jan Philipp</creatorcontrib><creatorcontrib>Ochs, Matthias</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Physical Education Index</collection><collection>Toxicology Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><jtitle>American journal of physiology. Lung cellular and molecular physiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Schneider, Jan Philipp</au><au>Ochs, Matthias</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Alterations of mouse lung tissue dimensions during processing for morphometry: a comparison of methods</atitle><jtitle>American journal of physiology. Lung cellular and molecular physiology</jtitle><addtitle>Am J Physiol Lung Cell Mol Physiol</addtitle><date>2014-02-15</date><risdate>2014</risdate><volume>306</volume><issue>4</issue><spage>L341</spage><epage>L350</epage><pages>L341-L350</pages><issn>1040-0605</issn><eissn>1522-1504</eissn><abstract>Preservation of original tissue dimensions is an essential prerequisite for morphometric studies. Shrinkage occurring during tissue processing for histology may severely influence the appearance of structures seen under the microscope and stereological calculations. Therefore, shrinkage has to be avoided so that estimates obtained by application of unbiased stereology are indeed unbiased. The present study investigates the alterations of tissue dimensions of mouse lung samples during processing for histology. Different fixatives as well as embedding protocols are considered. Mouse lungs were fixed by instillation of either 4% formalin or a mixture of 1.5% glutaraldehyde/1.5% formaldehyde. Tissue blocks were sampled according to principles of stereology for embedding in paraffin, glycol methacrylate without treatment with osmium tetroxide and uranyl acetate, and glycol methacrylate including treatment with osmium tetroxide and uranyl acetate before dehydration. Shrinkage was investigated by stereological measurements of dimensional changes of tissue cut faces. Results show a shrinkage of the cut face areas of roughly 40% per lung during paraffin embedding, 30% during "simple" glycol methacrylate embedding, and <3% during osmium tetroxide/uranyl acetate/glycol methacrylate embedding. Furthermore, the superiority of the glutaraldehyde-containing fixative regarding shrinkage is demonstrated. In conclusion, the use of a glutaraldehyde-containing fixative and embedding in glycol methacrylate with previous treatment of the samples with osmium tetroxide and uranyl acetate before dehydration is recommended for stereological studies of the mouse lung.</abstract><cop>United States</cop><pub>American Physiological Society</pub><pmid>24375800</pmid><doi>10.1152/ajplung.00329.2013</doi></addata></record> |
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| language | eng |
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| source | MEDLINE; American Physiological Society; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Animals Comparative analysis Desiccation Fixatives - chemistry Glutaral - chemistry Histology Lung - anatomy & histology Lungs Male Methacrylates - chemistry Mice Mice, Inbred C57BL Organ Size Organometallic Compounds - chemistry Osmium Tetroxide - chemistry Paraffin Embedding Physiology Plastic Embedding Rodents Specimen Handling Tissue Fixation Tissues |
| title | Alterations of mouse lung tissue dimensions during processing for morphometry: a comparison of methods |
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