2.3 | Image Processing
Image processing techniques were utilized to retrieve raw intensity
profiles from stacks of images for each respective treatment and their
duplicates during the 8-hour period of time-lapse fluorescence
microscopy. Raw red channel image stacks are processed using ImageJ
(National Institutes of Health, Bethesda, MD). First, contrast
enhancement is applied to obtain mitochondrial contours. This allows for
a ROI (region of interest) to be selected and applied over all 24 images
in a stack. For quantification, the selected ROI is used to retrieve
mitochondrial fluorescent intensity presented as A.F.U (arbitrary
fluorescent units) and background (dark regions) for background
subtraction. Intensity profiles are fitted to sigmoidal (Equation 1) and
linear (Equation 2) models using MATLAB R2022a (MathWorks Inc., Natick,
MA) to visualize trends in drug treatments.
\(y=\frac{1}{1+e^{-x}}\) (1)
\(y=mx+b\) (2)