4D mitochondrial biophysical parameters predict cell type in human organoid tissue

Mitochondria assume the form of a three-dimensional temporal network in the cell. There is a spectrum of mitochondrial network morphologies and dynamics that ranges from static, organized single mitochondria to branched, motile networks of connected mitochondria. This spectrum is sensitive to cell state, cell type, and organ system. Our hypothesis is that there are shared but also unique biophysical parameters that govern the mitochondrial temporal network dynamics in cells and tissues. We investigate two model systems, intestinal organoids and branching lung organoids, to extract and understand these biophysical parameters. We used adaptive optics-lattice light sheet microscopy (AO-LLSM) to capture 4D (x, y, z, time) mitochondria data in tissues with low photo-toxicity and high spatio-temporal resolution. Automated image processing and computational temporal network tracking was performed using the MitoTNT software package to quantify fission/fusion rates, morphological parameters, motility, and graph temporal network properties. We found unique temporal network signatures that correlate with cell and tissue type, supporting our hypothesis. Next steps include extending our findings to additional cell and tissue types.