Seahorse Biosciences 4 articles published in JoVE Biology Isolation of Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High Throughput Microplate Respiratory Measurements Nabil E. Boutagy1,2, Emily Pyne1, George W. Rogers3, Mostafa Ali1, Matthew W. Hulver1,2, Madlyn I. Frisard1,2 1The Department of Human Nutrition, Foods, and Exercise, Virginia Tech, 2The Metabolic Phenotyping Core, Virginia Tech, 3Seahorse Bioscience Here, we present a modification of a previously reported method that allows for the isolation of high quality and purified mitochondria from smaller quantities of mouse skeletal muscle. This procedure results in highly coupled mitochondria that respire with high function during microplate based respirometirc assays. Developmental Biology Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements Nabil E. Boutagy1,2, George W. Rogers3, Emily S. Pyne1, Mostafa M. Ali1, Matthew W. Hulver1,2, Madlyn I. Frisard1,2 1The Department of Human Nutrition, Foods, and Exercise, Virginia Tech, 2The Metabolic Phenotyping Core, Virginia Tech, 3Seahorse Bioscience The methods presented provide step-by-step instructions for the performance of a collection of microplate based respirometric assays using isolated mitochondria from minimal quantities of mouse skeletal muscle. These assays are able to measure mechanistic changes/adaptations in mitochondrial oxygen consumption in a commonly used animal model. Medicine Preparation and Respirometric Assessment of Mitochondria Isolated from Skeletal Muscle Tissue Obtained by Percutaneous Needle Biopsy Manish S. Bharadwaj1, Daniel J. Tyrrell1, Mary F. Lyles1, Jamehl L. Demons1, George W. Rogers2, Anthony J. A. Molina1 1Department of Internal Medicine, Section on Gerontology and Geriatric Medicine, Wake Forest School of Medicine, 2Seahorse Biosciences Methods for biopsy of Vastus lateralis, preparation of purified mitochondria, and respirometric profiling are described. The use of small muscle volume makes this technique suitable for clinical research applications. Biology Bioenergetic Profile Experiment using C2C12 Myoblast Cells David G. Nicholls1, Victor M. Darley-Usmar2, Min Wu3, Per Bo Jensen3, George W. Rogers3, David A. Ferrick3 1Buck Institute for Age Research, Novato, CA, 2Department of Pathology, Center for Free Radical Biology, University of Alabama at Birmingham - UAB, 3Seahorse Bioscience, North Billerica, MA A description of a method for profiling mitochondrial function in cells is provided. The mitochondrial profile generated provides four parameters of mitochondrial function that can be measured in one experiment: basal respiration rate, ATP-linked respiration, proton leak, and reserve capacity.