Tissue triage and freezing for models of skeletal muscle disease

Hui Meng, Paul M L Janssen, Robert W. Grange, Lin Yang, Alan H. Beggs, Lindsay C. Swanson, Stacy A. Cossette, Alison Frase, Martin K. Childers, Hendrikus "Henk" Granzier, Emanuela Gussoni, Michael W. Lawlor

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Skeletal 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.

Original languageEnglish (US)
JournalJournal of visualized experiments : JoVE
Issue number89
DOIs
StatePublished - 2014

Fingerprint

Triage
Freezing
Muscle
Skeletal Muscle
Tissue
Muscles
Congenital, Hereditary, and Neonatal Diseases and Abnormalities
Myoblasts
Muscular Diseases
Artifacts
Nitrogen
Cell Culture Techniques
Liquid nitrogen
Cell culture
Research
Proteins

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Meng, H., Janssen, P. M. L., Grange, R. W., Yang, L., Beggs, A. H., Swanson, L. C., ... Lawlor, M. W. (2014). Tissue triage and freezing for models of skeletal muscle disease. Journal of visualized experiments : JoVE, (89). https://doi.org/10.3791/51586

Tissue triage and freezing for models of skeletal muscle disease. / Meng, Hui; Janssen, Paul M L; Grange, Robert W.; Yang, Lin; Beggs, Alan H.; Swanson, Lindsay C.; Cossette, Stacy A.; Frase, Alison; Childers, Martin K.; Granzier, Hendrikus "Henk"; Gussoni, Emanuela; Lawlor, Michael W.

In: Journal of visualized experiments : JoVE, No. 89, 2014.

Research output: Contribution to journalArticle

Meng, H, Janssen, PML, Grange, RW, Yang, L, Beggs, AH, Swanson, LC, Cossette, SA, Frase, A, Childers, MK, Granzier, HH, Gussoni, E & Lawlor, MW 2014, 'Tissue triage and freezing for models of skeletal muscle disease', Journal of visualized experiments : JoVE, no. 89. https://doi.org/10.3791/51586
Meng, Hui ; Janssen, Paul M L ; Grange, Robert W. ; Yang, Lin ; Beggs, Alan H. ; Swanson, Lindsay C. ; Cossette, Stacy A. ; Frase, Alison ; Childers, Martin K. ; Granzier, Hendrikus "Henk" ; Gussoni, Emanuela ; Lawlor, Michael W. / Tissue triage and freezing for models of skeletal muscle disease. In: Journal of visualized experiments : JoVE. 2014 ; No. 89.
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