Mitochondrial Morphological Features Are Associated with Fission and Fusion Events
Figure 2
Quantification design for tracking mitochondrial morphology dynamics using mito_EYFP.
(A) Quantification schematic used to process images for analysis. Mitochondria were imaged in U2OS_mitoEYFP cells by fluorescent microscopy and were subjected to image processing that involved 2D deconvolution, top hat morphological transformation, intensity thresholding, and object quantification. (B) Intensity profiles from each individual cell were used to determine the thresholding boundaries for each image. Intensity profiles depict single distribution with a prominent right-hand elbow that we separated and termed distribution 1 and 2: pixels within distribution 1 are highlighted red and represent non-mitochondrial pixels whereas pixels within distribution 2 are highlighted green and represent mitochondrial pixels. (C) Split color of mitochondrial image in 1B highlighting location of pixels within each distribution. Mitochondrial measurements were performed with a threshold that excluded pixels in distribution 1.