IMCL Methods Draft

In this study, twenty-four young and older men and women participated. All subjects were matched for physical activity. Biopsies from the vastus lateralis were taken from each subject and imaged using electron microscopy. These images were provided by collaborators at the University of Vermont. Images were then analyzed using a protocol to identify intramyocellular lipid droplets within the muscle fiber. Lipid droplets were identified using the criteria as follows; a gray/white interior, somewhat circular in shape, contains a gray outline, doesn’t have multiple membranes on the outside or have debris inside, and usually adjacent to mitochondria. A protocol was then developed in order to create a thresholded image containing only lipid droplets. These thresholded images were then analyzed for circularity, area ratio, roundness, solidity, and % area. using ImageJ. This data was collected and cross-checked between multiple (2) undergraduate researchers.