Skeletal muscle triad junction ultrastructure by Focused-Ion-Beam milling of muscle and Cryo-Electron Tomography

Cryo-electron tomography (cryo-ET) has emerged as perhaps the only practical technique for revealing nanometer-level three-dimensional structural details of subcellular macromolecular complexes in their native context, inside the cell. As currently practiced, the specimen should be 0.1- 0.2 microns...

Full description

Saved in:
Bibliographic Details
Published in:European journal of translational myology 2015-01, Vol.25 (1), p.49-56
Main Authors: Wagenknecht, Terence, Hsieh, Chyongere, Marko, Michael
Format: Article
Language:English
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
cited_by
cites
container_end_page 56
container_issue 1
container_start_page 49
container_title European journal of translational myology
container_volume 25
creator Wagenknecht, Terence
Hsieh, Chyongere
Marko, Michael
description Cryo-electron tomography (cryo-ET) has emerged as perhaps the only practical technique for revealing nanometer-level three-dimensional structural details of subcellular macromolecular complexes in their native context, inside the cell. As currently practiced, the specimen should be 0.1- 0.2 microns in thickness to achieve optimal resolution. Thus, application of cryo-ET to intact frozen (vitreous) tissues, such as skeletal muscle, requires that they be sectioned. Cryo-ultramicrotomy is notoriously difficult and artifact-prone when applied to frozen cells and tissue, but a new technique, focused ion beam milling (cryo-FIB), shows great promise for “thinning” frozen biological specimens. Here we describe our initial results in applying cryo-FIB and cryo-ET to triad junctions of skeletal muscle.
doi_str_mv 10.4081/bam.2015.1.49
format article
fullrecord <record><control><sourceid>pubmedcentral_cross</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4454291</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>pubmedcentral_primary_oai_pubmedcentral_nih_gov_4454291</sourcerecordid><originalsourceid>FETCH-LOGICAL-c1329-e966ea8d554c12cebcf5320f55f6c075e69ce2ef4d99b368f17ce39b0a62764d3</originalsourceid><addsrcrecordid>eNpVkNFKwzAUhoMobsxdep8XaE3SJG1uBB2bDgQvnNchTU-3zrQpaSv07e2Yip6b_8DhfPB_CN1SEnOS0bvc1DEjVMQ05uoCzRlJ0ijlklz-2Wdo2XVHMk1GpaLpNZoxSQTjlM9R-_YBDnrjcD101gHuQ2UKfBwa21e-wYPrg-n6MNh-CIDzEW-8HToooq1vokcwNa4r56pmj335wzBNgVdh9NHage3DhNn52u-DaQ_jDboqjetg-Z0L9L5Z71bP0cvr03b18BJZmjAVgZISTFYIwS1lFnJbioSRUohSWpIKkMoCg5IXSuWJzEqaWkhUToxkqeRFskD3Z2475DUUFpqpiNNtqGoTRu1Npf9fmuqg9_5Tcy44U3QCRGeADb7rApS_v5Tok3092dcn-5pqrpIv2YN6Dw</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype></control><display><type>article</type><title>Skeletal muscle triad junction ultrastructure by Focused-Ion-Beam milling of muscle and Cryo-Electron Tomography</title><source>PubMed Central (Open Access)</source><source>Alma/SFX Local Collection</source><creator>Wagenknecht, Terence ; Hsieh, Chyongere ; Marko, Michael</creator><creatorcontrib>Wagenknecht, Terence ; Hsieh, Chyongere ; Marko, Michael</creatorcontrib><description>Cryo-electron tomography (cryo-ET) has emerged as perhaps the only practical technique for revealing nanometer-level three-dimensional structural details of subcellular macromolecular complexes in their native context, inside the cell. As currently practiced, the specimen should be 0.1- 0.2 microns in thickness to achieve optimal resolution. Thus, application of cryo-ET to intact frozen (vitreous) tissues, such as skeletal muscle, requires that they be sectioned. Cryo-ultramicrotomy is notoriously difficult and artifact-prone when applied to frozen cells and tissue, but a new technique, focused ion beam milling (cryo-FIB), shows great promise for “thinning” frozen biological specimens. Here we describe our initial results in applying cryo-FIB and cryo-ET to triad junctions of skeletal muscle.</description><identifier>ISSN: 2037-7460</identifier><identifier>ISSN: 2037-7452</identifier><identifier>EISSN: 2037-7460</identifier><identifier>DOI: 10.4081/bam.2015.1.49</identifier><identifier>PMID: 26052414</identifier><language>eng</language><ispartof>European journal of translational myology, 2015-01, Vol.25 (1), p.49-56</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4454291/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4454291/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids></links><search><creatorcontrib>Wagenknecht, Terence</creatorcontrib><creatorcontrib>Hsieh, Chyongere</creatorcontrib><creatorcontrib>Marko, Michael</creatorcontrib><title>Skeletal muscle triad junction ultrastructure by Focused-Ion-Beam milling of muscle and Cryo-Electron Tomography</title><title>European journal of translational myology</title><description>Cryo-electron tomography (cryo-ET) has emerged as perhaps the only practical technique for revealing nanometer-level three-dimensional structural details of subcellular macromolecular complexes in their native context, inside the cell. As currently practiced, the specimen should be 0.1- 0.2 microns in thickness to achieve optimal resolution. Thus, application of cryo-ET to intact frozen (vitreous) tissues, such as skeletal muscle, requires that they be sectioned. Cryo-ultramicrotomy is notoriously difficult and artifact-prone when applied to frozen cells and tissue, but a new technique, focused ion beam milling (cryo-FIB), shows great promise for “thinning” frozen biological specimens. Here we describe our initial results in applying cryo-FIB and cryo-ET to triad junctions of skeletal muscle.</description><issn>2037-7460</issn><issn>2037-7452</issn><issn>2037-7460</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><recordid>eNpVkNFKwzAUhoMobsxdep8XaE3SJG1uBB2bDgQvnNchTU-3zrQpaSv07e2Yip6b_8DhfPB_CN1SEnOS0bvc1DEjVMQ05uoCzRlJ0ijlklz-2Wdo2XVHMk1GpaLpNZoxSQTjlM9R-_YBDnrjcD101gHuQ2UKfBwa21e-wYPrg-n6MNh-CIDzEW-8HToooq1vokcwNa4r56pmj335wzBNgVdh9NHage3DhNn52u-DaQ_jDboqjetg-Z0L9L5Z71bP0cvr03b18BJZmjAVgZISTFYIwS1lFnJbioSRUohSWpIKkMoCg5IXSuWJzEqaWkhUToxkqeRFskD3Z2475DUUFpqpiNNtqGoTRu1Npf9fmuqg9_5Tcy44U3QCRGeADb7rApS_v5Tok3092dcn-5pqrpIv2YN6Dw</recordid><startdate>20150115</startdate><enddate>20150115</enddate><creator>Wagenknecht, Terence</creator><creator>Hsieh, Chyongere</creator><creator>Marko, Michael</creator><scope>AAYXX</scope><scope>CITATION</scope><scope>5PM</scope></search><sort><creationdate>20150115</creationdate><title>Skeletal muscle triad junction ultrastructure by Focused-Ion-Beam milling of muscle and Cryo-Electron Tomography</title><author>Wagenknecht, Terence ; Hsieh, Chyongere ; Marko, Michael</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c1329-e966ea8d554c12cebcf5320f55f6c075e69ce2ef4d99b368f17ce39b0a62764d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wagenknecht, Terence</creatorcontrib><creatorcontrib>Hsieh, Chyongere</creatorcontrib><creatorcontrib>Marko, Michael</creatorcontrib><collection>CrossRef</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>European journal of translational myology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wagenknecht, Terence</au><au>Hsieh, Chyongere</au><au>Marko, Michael</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Skeletal muscle triad junction ultrastructure by Focused-Ion-Beam milling of muscle and Cryo-Electron Tomography</atitle><jtitle>European journal of translational myology</jtitle><date>2015-01-15</date><risdate>2015</risdate><volume>25</volume><issue>1</issue><spage>49</spage><epage>56</epage><pages>49-56</pages><issn>2037-7460</issn><issn>2037-7452</issn><eissn>2037-7460</eissn><abstract>Cryo-electron tomography (cryo-ET) has emerged as perhaps the only practical technique for revealing nanometer-level three-dimensional structural details of subcellular macromolecular complexes in their native context, inside the cell. As currently practiced, the specimen should be 0.1- 0.2 microns in thickness to achieve optimal resolution. Thus, application of cryo-ET to intact frozen (vitreous) tissues, such as skeletal muscle, requires that they be sectioned. Cryo-ultramicrotomy is notoriously difficult and artifact-prone when applied to frozen cells and tissue, but a new technique, focused ion beam milling (cryo-FIB), shows great promise for “thinning” frozen biological specimens. Here we describe our initial results in applying cryo-FIB and cryo-ET to triad junctions of skeletal muscle.</abstract><pmid>26052414</pmid><doi>10.4081/bam.2015.1.49</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 2037-7460
ispartof European journal of translational myology, 2015-01, Vol.25 (1), p.49-56
issn 2037-7460
2037-7452
2037-7460
language eng
recordid cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4454291
source PubMed Central (Open Access); Alma/SFX Local Collection
title Skeletal muscle triad junction ultrastructure by Focused-Ion-Beam milling of muscle and Cryo-Electron Tomography
url http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-08T05%3A01%3A45IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-pubmedcentral_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Skeletal%20muscle%20triad%20junction%20ultrastructure%20by%20Focused-Ion-Beam%20milling%20of%20muscle%20and%20Cryo-Electron%20Tomography&rft.jtitle=European%20journal%20of%20translational%20myology&rft.au=Wagenknecht,%20Terence&rft.date=2015-01-15&rft.volume=25&rft.issue=1&rft.spage=49&rft.epage=56&rft.pages=49-56&rft.issn=2037-7460&rft.eissn=2037-7460&rft_id=info:doi/10.4081/bam.2015.1.49&rft_dat=%3Cpubmedcentral_cross%3Epubmedcentral_primary_oai_pubmedcentral_nih_gov_4454291%3C/pubmedcentral_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c1329-e966ea8d554c12cebcf5320f55f6c075e69ce2ef4d99b368f17ce39b0a62764d3%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_id=info:pmid/26052414&rfr_iscdi=true