Data from: Diffusive spreading across dynamic mitochondrial network architectures
Data files
Feb 04, 2026 version files 56.63 GB
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Holt_et_al_live_imaging.zip
56.63 GB
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README.md
3.38 KB
Abstract
Networks of physical units can vary from a stationary set of spatially-embedded links to a collection of mobile agents that undergo transient social interactions. In living cells, mitochondria form architectures that span across these regimes, transitioning between fragmented, partly connected, and highly fused structures depending on cell type and state. Diffusive transport of biomolecular components through these networks limits the heterogeneity of the mitochondrial population. Here we address the connection between dynamic network architecture and the rate of diffusive mixing through simulations and analytic models that incorporate fusion, fission, and rearrangement. We find that the material delivered from a source to the rest of the network depends on the network dimensionality and a balance of competing timescales for encounter, fusion, and diffusive dispersion. We extract morphological and dynamic parameters for mitochondrial networks in three human cell lines, demonstrating that different cells span across both the physical and social network regimes. Mixing in mitochondrial networks is shown to be limited by material transport through connected tubules for slowly-diffusing particles and by inter-mitochondrial encounter rates for rapidly diffusing ones. These results provide a quantitative basis for predicting the homogenization of proteins, lipids, ions, or genetic material through the mitochondrial population. The general principles identified in this work capture diffusive spreading through both social and physical networks, unifying a continuum of spatial network architectures. These data comprise Airyscan fluorescence micrographs of mitochondrial network dynamics in human fibroblast, osteosarcoma, and neuroblastoma cells labeled with Mitotracker dye and SYBR Gold, and accompanying capture metadata.
Dataset DOI: 10.5061/dryad.cjsxksnkb
Description of the dataset
Dataset Overview
For a comprehensive overview and the methodology used to generate and process all the data in this repository, please see the corresponding preprint at https://arxiv.org/abs/2506.05643.
Notes for Files
Microscopy and Image Acquisition
All images were acquired using a Zeiss LSM 980 with Airyscan 2 laser scanning confocal microscope, equipped with 405-, 488-, 561-, and 639-nm laser lines and Fast Airyscan detector array. Images were acquired using an inverted 63×/1.4 numerical aperture oil objective. All live imaging was done in a humidified chamber at 37°C and in the presence of 5% CO2. Airyscan processing was performed using Zeiss ZEN Blue software version 3.7 (Carl Zeiss).
Holt_et_al_live_imaging.zip
The compressed master directory contains 3 sub-directories corresponding to the three cell types imaged for the study, including all time-lapse microscopy analyzed and representative image shown in the main figures.
File Names
File names follow the pattern:
(celltype)_(fluorescent protein or dye(s))_(passage number).ome.tiff
Movie file abbreviations:
pN: where N equals passage number of cell culture at time of image acquisition
mtdr: MitoTracker Deep Red (Thermo Fisher Scientific Catalog No. M22426)
sybr: SYBRGold (Thermo Fisher Scientific catalog no. S11494)
imr90: Fibroblast (IMR90)
sy5y: Neuroblastoma (SH-SY5Y)
u2os: Osteosarcoma Cas9-expressing (U2OS Cas9)
NT: no treatment
File Types
- .ome.tiff: Zeiss Airyscan processed microscopy time-lapse video.
- .txt: plain text file containing image metadata.
Description of Directories and File Structure
FILENAME_Data
|- IMR90/
|---MitoTracker_SYBRGold
|- Neuroblastoma/
|---MitoTracker_SYBRGold
|---Mito-eGFP
|- U2OS/
|---Mito-BFP
|---MitoTracker
Sub-directories
#### *IMR90*
|---MitoTracker_SYBRGold
This folder contains all 3D live-cell fluorescence microscopy timelapse videos of mitochondrial networks in fibroblasts (IMR90). Each movie is provided as a .ome.tiff file along with a corresponding text file containing acquisition metadata. Images are matched to corresponding metadata by a shared capture number in the file name.
#### *Neuroblastoma*
|---MitoTracker_SYBRGold
|---Mito-eGFP
This folder contains all 3D live-cell fluorescence microscopy timelapse videos of mitochondrial networks in neuroblastoma cells (SH-SY5Y). Each movie is provided as a .ome.tiff file along with a corresponding text file containing acquisition metadata. Images are matched to corresponding metadata by a shared capture number in the file name.
#### *U2OS*
|---Mito-BFP
|---MitoTracker
This folder contains all 3D live-cell fluorescence microscopy timelapse videos of mitochondrial networks in osteosarcoma cells (U2OS). Each movie is provided as a .ome.tiff file along with a corresponding text file containing acquisition metadata. Images are matched to corresponding metadata by a shared capture number in the file name.
Code/Software
Airyscan processing was performed using Zeiss ZEN Blue software version 3.7 (Carl Zeiss), RRID:SCR_013672.
