Cellsens

Hypodermal mitochondria were labeled by staining with MitoTracker Red CMXRos (Invitrogen) and observed using Olympus BX51 microscope equipped for Nomarski and fluorescence microscopy. Young adult worms were either stained in MitoTracker solution for 20 minutes or fed with MitoTracker treated OP50 for 30 minutes. After staining, worms were destained for 20 minutes. Stained worms were mounted as described [49 (link)] and observed using Olympus BX51 microscope equipped for Nomarski and fluorescence microscopy. The width of the mitochondria containing region was determined using the Olympus cellSens, ImageJ or Photoshop (Adobe).
Body wall muscle mitochondria were labeled by mitochondria-targeted green fluorescent protein (mtGFP) expressed in the body wall muscle (zcIs14) [43 (link)]. Using the ImageJ software, from a fluorescence image, an outline of a single muscle cell was selected. The area covered by mitochondria was determined by thresholding the pixel intensity, and the percentage of area covered by mitochondria (as a fraction of the total muscle area) was calculated.

All images were acquired using an Olympus FluoView FV3000 confocal microscope. Image processing was performed using Olympus CellSens, Adobe Photoshop, and Adobe Illustrator. The co-localization study was performed with the co-localization threshold tool using the ImageJ software. The software allowed both the calculation of the Pearson’s correlation coefficient between two channels and their relative percentage of co-localization. Sections from 3 human brains were utilized for cell quantification.

Images were acquired in Olympus FV1200 confocal microscope and Olympus cellSens, treated with Adobe Photoshop CS6 image programs. Wing size and space between vein 3 and vein 4 were measured on each picture using the ImageJ computer program.