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Combining quantitative 2D and 3D image analysis in the serial block face SEM: application to secretory organelles of pancreatic islet cells
Summary A combination of two‐dimensional (2D) and three‐dimensional (3D) analyses of tissue volume ultrastructure acquired by serial block face scanning electron microscopy can greatly shorten the time required to obtain quantitative information from big data sets that contain many billions of voxel...
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| Published in: | Journal of microscopy (Oxford) 2015-08, Vol.259 (2), p.155-164 |
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| Main Authors: | , , , , , , , |
| Format: | Article |
| Language: | English |
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| cited_by | cdi_FETCH-LOGICAL-c5796-390dc83520d50e42dcfd9e3ddfebf97f0bcdd59199077b98e8141751d55988d43 |
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| cites | cdi_FETCH-LOGICAL-c5796-390dc83520d50e42dcfd9e3ddfebf97f0bcdd59199077b98e8141751d55988d43 |
| container_end_page | 164 |
| container_issue | 2 |
| container_start_page | 155 |
| container_title | Journal of microscopy (Oxford) |
| container_volume | 259 |
| creator | SHOMORONY, A. PFEIFER, C.R. ARONOVA, M.A. ZHANG, G. CAI, T. XU, H. NOTKINS, A.L. LEAPMAN, R.D. |
| description | Summary
A combination of two‐dimensional (2D) and three‐dimensional (3D) analyses of tissue volume ultrastructure acquired by serial block face scanning electron microscopy can greatly shorten the time required to obtain quantitative information from big data sets that contain many billions of voxels. Thus, to analyse the number of organelles of a specific type, or the total volume enclosed by a population of organelles within a cell, it is possible to estimate the number density or volume fraction of that organelle using a stereological approach to analyse randomly selected 2D block face views through the cells, and to combine such estimates with precise measurement of 3D cell volumes by delineating the plasma membrane in successive block face images. The validity of such an approach can be easily tested since the entire 3D tissue volume is available in the serial block face scanning electron microscopy data set. We have applied this hybrid 3D/2D technique to determine the number of secretory granules in the endocrine α and β cells of mouse pancreatic islets of Langerhans, and have been able to estimate the total insulin content of a β cell. |
| doi_str_mv | 10.1111/jmi.12276 |
| format | article |
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A combination of two‐dimensional (2D) and three‐dimensional (3D) analyses of tissue volume ultrastructure acquired by serial block face scanning electron microscopy can greatly shorten the time required to obtain quantitative information from big data sets that contain many billions of voxels. Thus, to analyse the number of organelles of a specific type, or the total volume enclosed by a population of organelles within a cell, it is possible to estimate the number density or volume fraction of that organelle using a stereological approach to analyse randomly selected 2D block face views through the cells, and to combine such estimates with precise measurement of 3D cell volumes by delineating the plasma membrane in successive block face images. The validity of such an approach can be easily tested since the entire 3D tissue volume is available in the serial block face scanning electron microscopy data set. We have applied this hybrid 3D/2D technique to determine the number of secretory granules in the endocrine α and β cells of mouse pancreatic islets of Langerhans, and have been able to estimate the total insulin content of a β cell.</description><identifier>ISSN: 0022-2720</identifier><identifier>EISSN: 1365-2818</identifier><identifier>DOI: 10.1111/jmi.12276</identifier><identifier>PMID: 26139222</identifier><identifier>CODEN: JMICAR</identifier><language>eng</language><publisher>England: Wiley Subscription Services, Inc</publisher><subject>Animals ; Glucagon-Secreting Cells - ultrastructure ; Imaging, Three-Dimensional ; Insulin - analysis ; Insulin-Secreting Cells - chemistry ; Insulin-Secreting Cells - ultrastructure ; Male ; Mice ; Microscopy, Electron, Scanning - methods ; Pancreas ; pancreatic islets ; Rodents ; Scanning electron microscopy ; secretory granules ; Secretory Vesicles - ultrastructure ; serial block face SEM ; stereology ; Themed Issue Paper ; Themed Issue Papers ; α cells ; β cells</subject><ispartof>Journal of microscopy (Oxford), 2015-08, Vol.259 (2), p.155-164</ispartof><rights>2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society</rights><rights>2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.</rights><rights>Journal compilation © 2015 Royal Microscopical Society</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5796-390dc83520d50e42dcfd9e3ddfebf97f0bcdd59199077b98e8141751d55988d43</citedby><cites>FETCH-LOGICAL-c5796-390dc83520d50e42dcfd9e3ddfebf97f0bcdd59199077b98e8141751d55988d43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26139222$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>SHOMORONY, A.</creatorcontrib><creatorcontrib>PFEIFER, C.R.</creatorcontrib><creatorcontrib>ARONOVA, M.A.</creatorcontrib><creatorcontrib>ZHANG, G.</creatorcontrib><creatorcontrib>CAI, T.</creatorcontrib><creatorcontrib>XU, H.</creatorcontrib><creatorcontrib>NOTKINS, A.L.</creatorcontrib><creatorcontrib>LEAPMAN, R.D.</creatorcontrib><title>Combining quantitative 2D and 3D image analysis in the serial block face SEM: application to secretory organelles of pancreatic islet cells</title><title>Journal of microscopy (Oxford)</title><addtitle>J Microsc</addtitle><description>Summary
A combination of two‐dimensional (2D) and three‐dimensional (3D) analyses of tissue volume ultrastructure acquired by serial block face scanning electron microscopy can greatly shorten the time required to obtain quantitative information from big data sets that contain many billions of voxels. Thus, to analyse the number of organelles of a specific type, or the total volume enclosed by a population of organelles within a cell, it is possible to estimate the number density or volume fraction of that organelle using a stereological approach to analyse randomly selected 2D block face views through the cells, and to combine such estimates with precise measurement of 3D cell volumes by delineating the plasma membrane in successive block face images. The validity of such an approach can be easily tested since the entire 3D tissue volume is available in the serial block face scanning electron microscopy data set. We have applied this hybrid 3D/2D technique to determine the number of secretory granules in the endocrine α and β cells of mouse pancreatic islets of Langerhans, and have been able to estimate the total insulin content of a β cell.</description><subject>Animals</subject><subject>Glucagon-Secreting Cells - ultrastructure</subject><subject>Imaging, Three-Dimensional</subject><subject>Insulin - analysis</subject><subject>Insulin-Secreting Cells - chemistry</subject><subject>Insulin-Secreting Cells - ultrastructure</subject><subject>Male</subject><subject>Mice</subject><subject>Microscopy, Electron, Scanning - methods</subject><subject>Pancreas</subject><subject>pancreatic islets</subject><subject>Rodents</subject><subject>Scanning electron microscopy</subject><subject>secretory granules</subject><subject>Secretory Vesicles - ultrastructure</subject><subject>serial block face SEM</subject><subject>stereology</subject><subject>Themed Issue Paper</subject><subject>Themed Issue Papers</subject><subject>α cells</subject><subject>β cells</subject><issn>0022-2720</issn><issn>1365-2818</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><recordid>eNp1kc1u1DAURi0EokNhwQsgS2xgkdY_cWKzQELTAkWtWABry7Fvph6cOLWTonkGXhq3UypAwhvL-o6O7vWH0HNKjmg5x9vBH1HG2uYBWlHeiIpJKh-iFSGMVaxl5AA9yXlLCJFCksfogDWUK8bYCv1cx6Hzox83-Gox4-xnM_trwOwEm9FhfoL9YDZQHibsss_Yj3i-BJwheRNwF6L9jntjAX85vXiDzTQFb4siFiwWyiaYY9rhmDZmhBAg49jjyYwlKJjFPgeYsS1Rfooe9SZkeHZ3H6Jv70-_rj9W558_nK3fnVdWtKqpuCLOSi4YcYJAzZztnQLuXA9dr9qedNY5oahSpG07JUHSmraCOiGUlK7mh-jt3jst3QDOwjgnE_SUyqppp6Px-u9k9Jd6E691LaioOS-CV3eCFK8WyLMefL5ZoawYl6xpoyThgkhV0Jf_oNu4pPKZt1RLVV2KK9TrPWVTzDlBfz8MJfqmYl0q1rcVF_bFn9Pfk787LcDxHvjhA-z-b9KfLs72yl8iQbHq</recordid><startdate>201508</startdate><enddate>201508</enddate><creator>SHOMORONY, A.</creator><creator>PFEIFER, C.R.</creator><creator>ARONOVA, M.A.</creator><creator>ZHANG, G.</creator><creator>CAI, T.</creator><creator>XU, H.</creator><creator>NOTKINS, A.L.</creator><creator>LEAPMAN, R.D.</creator><general>Wiley Subscription Services, Inc</general><general>John Wiley and Sons Inc</general><scope>24P</scope><scope>WIN</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>7U5</scope><scope>8FD</scope><scope>L7M</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>201508</creationdate><title>Combining quantitative 2D and 3D image analysis in the serial block face SEM: application to secretory organelles of pancreatic islet cells</title><author>SHOMORONY, A. ; PFEIFER, C.R. ; ARONOVA, M.A. ; ZHANG, G. ; CAI, T. ; XU, H. ; NOTKINS, A.L. ; LEAPMAN, R.D.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5796-390dc83520d50e42dcfd9e3ddfebf97f0bcdd59199077b98e8141751d55988d43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Animals</topic><topic>Glucagon-Secreting Cells - ultrastructure</topic><topic>Imaging, Three-Dimensional</topic><topic>Insulin - analysis</topic><topic>Insulin-Secreting Cells - chemistry</topic><topic>Insulin-Secreting Cells - ultrastructure</topic><topic>Male</topic><topic>Mice</topic><topic>Microscopy, Electron, Scanning - methods</topic><topic>Pancreas</topic><topic>pancreatic islets</topic><topic>Rodents</topic><topic>Scanning electron microscopy</topic><topic>secretory granules</topic><topic>Secretory Vesicles - ultrastructure</topic><topic>serial block face SEM</topic><topic>stereology</topic><topic>Themed Issue Paper</topic><topic>Themed Issue Papers</topic><topic>α cells</topic><topic>β cells</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>SHOMORONY, A.</creatorcontrib><creatorcontrib>PFEIFER, C.R.</creatorcontrib><creatorcontrib>ARONOVA, M.A.</creatorcontrib><creatorcontrib>ZHANG, G.</creatorcontrib><creatorcontrib>CAI, T.</creatorcontrib><creatorcontrib>XU, H.</creatorcontrib><creatorcontrib>NOTKINS, A.L.</creatorcontrib><creatorcontrib>LEAPMAN, R.D.</creatorcontrib><collection>Wiley Online Library Open Access</collection><collection>Wiley Online Library Free Content</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Technology Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of microscopy (Oxford)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>SHOMORONY, A.</au><au>PFEIFER, C.R.</au><au>ARONOVA, M.A.</au><au>ZHANG, G.</au><au>CAI, T.</au><au>XU, H.</au><au>NOTKINS, A.L.</au><au>LEAPMAN, R.D.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Combining quantitative 2D and 3D image analysis in the serial block face SEM: application to secretory organelles of pancreatic islet cells</atitle><jtitle>Journal of microscopy (Oxford)</jtitle><addtitle>J Microsc</addtitle><date>2015-08</date><risdate>2015</risdate><volume>259</volume><issue>2</issue><spage>155</spage><epage>164</epage><pages>155-164</pages><issn>0022-2720</issn><eissn>1365-2818</eissn><coden>JMICAR</coden><abstract>Summary
A combination of two‐dimensional (2D) and three‐dimensional (3D) analyses of tissue volume ultrastructure acquired by serial block face scanning electron microscopy can greatly shorten the time required to obtain quantitative information from big data sets that contain many billions of voxels. Thus, to analyse the number of organelles of a specific type, or the total volume enclosed by a population of organelles within a cell, it is possible to estimate the number density or volume fraction of that organelle using a stereological approach to analyse randomly selected 2D block face views through the cells, and to combine such estimates with precise measurement of 3D cell volumes by delineating the plasma membrane in successive block face images. The validity of such an approach can be easily tested since the entire 3D tissue volume is available in the serial block face scanning electron microscopy data set. We have applied this hybrid 3D/2D technique to determine the number of secretory granules in the endocrine α and β cells of mouse pancreatic islets of Langerhans, and have been able to estimate the total insulin content of a β cell.</abstract><cop>England</cop><pub>Wiley Subscription Services, Inc</pub><pmid>26139222</pmid><doi>10.1111/jmi.12276</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Journal of microscopy (Oxford), 2015-08, Vol.259 (2), p.155-164 |
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| subjects | Animals Glucagon-Secreting Cells - ultrastructure Imaging, Three-Dimensional Insulin - analysis Insulin-Secreting Cells - chemistry Insulin-Secreting Cells - ultrastructure Male Mice Microscopy, Electron, Scanning - methods Pancreas pancreatic islets Rodents Scanning electron microscopy secretory granules Secretory Vesicles - ultrastructure serial block face SEM stereology Themed Issue Paper Themed Issue Papers α cells β cells |
| title | Combining quantitative 2D and 3D image analysis in the serial block face SEM: application to secretory organelles of pancreatic islet cells |
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