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Tissue Triage and Freezing for Models of Skeletal Muscle Disease

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dc.contributor.authorMeng, Huien
dc.contributor.authorJanssen, Paul M. L.en
dc.contributor.authorGrange, Robert W.en
dc.contributor.authorYang, Linen
dc.contributor.authorBeggs, Alan H.en
dc.contributor.authorSwansons, Lindsay C.en
dc.contributor.authorCossette, Stacy A.en
dc.contributor.authorFrase, Alisonen
dc.contributor.authorChilders, Martin K.en
dc.contributor.authorGranzier, Henken
dc.contributor.authorGussoni, Emanuelaen
dc.contributor.authorLawlor, Michael W.en
dc.date.accessioned2020-01-21T14:31:29Zen
dc.date.available2020-01-21T14:31:29Zen
dc.date.issued2014-07-01en
dc.date.updated2020-01-21T14:31:22Zen
dc.description.abstractSkeletal muscle is a unique tissue because of its structure and function, which requires specific protocols for tissue collection to obtain optimal results from functional, cellular, molecular, and pathological evaluations. Due to the subtlety of some pathological abnormalities seen in congenital muscle disorders and the potential for fixation to interfere with the recognition of these features, pathological evaluation of frozen muscle is preferable to fixed muscle when evaluating skeletal muscle for congenital muscle disease. Additionally, the potential to produce severe freezing artifacts in muscle requires specific precautions when freezing skeletal muscle for histological examination that are not commonly used when freezing other tissues. This manuscript describes a protocol for rapid freezing of skeletal muscle using isopentane (2-methylbutane) cooled with liquid nitrogen to preserve optimal skeletal muscle morphology. This procedure is also effective for freezing tissue intended for genetic or protein expression studies. Furthermore, we have integrated our freezing protocol into a broader procedure that also describes preferred methods for the short term triage of tissue for (1) single fiber functional studies and (2) myoblast cell culture, with a focus on the minimum effort necessary to collect tissue and transport it to specialized research or reference labs to complete these studies. Overall, this manuscript provides an outline of how fresh tissue can be effectively distributed for a variety of phenotypic studies and thereby provides standard operating procedures (SOPs) for pathological studies related to congenital muscle disease.en
dc.description.versionPublished versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.3791/51586en
dc.identifier.eissn1940-087Xen
dc.identifier.issn1940-087Xen
dc.identifier.orcidGrange, Robert [0000-0002-8288-2793]en
dc.identifier.urihttp://hdl.handle.net/10919/96524en
dc.language.isoenen
dc.publisherJournal of Visualized Experimentsen
dc.relation.urihttp://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000349296100043&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1en
dc.rightsCreative Commons Attribution-NonCommercial-NoDerivatives 3.0 Unporteden
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/en
dc.subjecttissueen
dc.subjectfreezingen
dc.subjectmuscleen
dc.subjectisopentaneen
dc.subjectpathologyen
dc.subjectfunctional testingen
dc.subjectcell cultureen
dc.subjectfiber-type disproportionen
dc.subjectthin filament lengthen
dc.subjectlife-spanen
dc.subjectmyoblastsen
dc.subjectmutationsen
dc.subjectmyopathyen
dc.subjectweaknessen
dc.subjecttherapyen
dc.subjectcommonen
dc.subjecttpm3en
dc.subjectbasic protocolen
dc.subject.meshMuscle, Skeletalen
dc.subject.meshAnimalsen
dc.subject.meshMiceen
dc.subject.meshMuscular Diseasesen
dc.subject.meshDisease Models, Animalen
dc.subject.meshCell Culture Techniquesen
dc.subject.meshTissue Fixationen
dc.subject.meshPhenotypeen
dc.subject.meshFreezingen
dc.titleTissue Triage and Freezing for Models of Skeletal Muscle Diseaseen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciencesen
pubs.organisational-group/Virginia Tech/Faculty of Health Sciencesen
pubs.organisational-group/Virginia Tech/All T&R Facultyen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/Human Nutrition, Foods, & Exerciseen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/CALS T&R Facultyen
pubs.organisational-group/Virginia Tech/University Research Institutes/Fralin Life Sciences/Durelle Scotten
pubs.organisational-group/Virginia Techen
pubs.organisational-group/Virginia Tech/University Research Institutes/Fralin Life Sciencesen
pubs.organisational-group/Virginia Tech/University Research Institutesen

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