title: Tissue triage and freezing for models of skeletal muscle disease.
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.
authors: Division of Pediatric Pathology, Department of Pathology and Laboratory Medicine, Medical College of Wisconsin.
authors: Hui
authors: H
authors: Meng
authors: Department of Physiology and Cell Biology, The Ohio State University.
authors: Paul M L
authors: PM
authors: Janssen
authors: Department of Human Nutrition, Foods and Exercise, Virginia Tech.
authors: Robert W
authors: RW
authors: Grange
authors: Division of Biomedical Informatics, Department of Biostatistics, Department of Computer Science, University of Kentucky.
authors: Lin
authors: L
authors: Yang
authors: Division of Genetics and Genomics, The Manton Center for Orphan Disease Research, Boston Children's Hospital, Harvard Medical School.
authors: Alan H
authors: AH
authors: Beggs
authors: Division of Genetics and Genomics, The Manton Center for Orphan Disease Research, Boston Children's Hospital, Harvard Medical School.
authors: Lindsay C
authors: LC
authors: Swanson
authors: Division of Pediatric Pathology, Department of Pathology and Laboratory Medicine, Medical College of Wisconsin; Cure Congenital Muscular Dystrophy.
authors: Stacy A
authors: SA
authors: Cossette
authors: Joshua Frase Foundation.
authors: Alison
authors: A
authors: Frase
authors: Department of Rehabilitation Medicine, University of Washington.
authors: Martin K
authors: MK
authors: Childers
authors: Department of Physiology, University of Arizona.
authors: Henk
authors: H
authors: Granzier
authors: Division of Genetics and Genomics, The Manton Center for Orphan Disease Research, Boston Children's Hospital, Harvard Medical School.
authors: Emanuela
authors: E
authors: Gussoni
authors: Division of Pediatric Pathology, Department of Pathology and Laboratory Medicine, Medical College of Wisconsin; mlawlor@mcw.edu.
authors: Michael W
authors: MW
authors: Lawlor
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D004195:Disease Models, Animal; D005615:Freezing; D051379:Mice; D018482:Muscle, Skeletal / Q000033:anatomy & histology* / Q000473:pathology / Q000502:physiology*; D009135:Muscular Diseases / Q000151:congenital* / Q000473:pathology*; D010641:Phenotype; D016707:Tissue Fixation / Q000379:methods*