Automatically Quantify the Dynamics of Live Mitochondrial Assays

Nanolive launches another ground-breaking tool in cell analysis. Powered by AI, the Smart Mitochondrial AssayLIVE, the world’s first fully automated solution for non-invasive, quantitative assessment of mitochondrial dynamics at the individual and network levels. First reported in Nature Communications (June 2024), this label-free system is now commercially available as a single-click digital assay,

Created to accelerate drug discovery, the Smart Mitochondrial AssayLIVE provides deeper insights into mitochondrial responses without any need for dyes or complex sample preparation. This avoids any potential unwanted interations caused by the use of traditional markers which may lead to biasing or limitations with imaging.

Key capabilities include:

  • One click assay
  • Time-lapse imaging without phototoxicity
  • Capture mitochondrial responses
  • Quantify morphological changes at the mitochondrian and network levels
  • Correlate fluorescence and functional analyses

Comprehensive metric are generated by Nanolive’s holotomographic images by automatically segmenting mitochondria to produce more than 30 label-free metrics per mitochondria and per cell.

These include mitochondrial:

  • Count
  • Radius
  • Length
  • Dry mass
  • Dry mass density
  • Number of branches
  • Number of endpoints
  • Number of junctions

These comprehensive, dye-free insights are of great interest to biopharma researchers as they allow them to assess critical mitochondrial dynamics, such as fission, fusion, swelling, mitophagy, and network complexity, key for evaluating metabolic function, cellular stress, and drug-induced toxicity with unmatched precision.

Nanolive Smart Mitochondrial Assay LIVE
Label-free mitochondrial segmentation in HeLa cells. Nanolive’s refractive index holotomographic images of HeLa cells at the population level, with a zoomed-in view of a single cell. Refractive index image alone (left) and with digital mitochondrial overlays (centre) following analysis performed with the Smart Mitochondrial AssayLIVE. On the right we display an analysis output showing the relationship between mitochondrial dry mass, count, and time. Both metrics decrease in tandem, confirming that mitochondria within the cells are being progressively degraded.

For an even deeper level of understanding, the label-free data from the Smart Mitochondrial AssayLIVE can be multiplexed with optional fluorescence imaging, visualisation, and quantification. Using automated analysis in the EVE Explorer Pro software enables:

  • Correlative fluorescence imaging with refractive index-based structures, to associate protein expression with morphological properties for deeper insights
  • Definition of distinct cell sub-populations in co-cultures by fluorescence signal, enabling comparison of responses across specific cell types
  • Quantification of fluorescence changes over time allowing for the monitoring of dynamic functional responses, such as ROS production, to evaluate drug efficacy and toxicity

The Smart Mitochondrial AssayLIVE system has applications in areas such as:

  • Oncology – to evaluate the efficacy of mitochondrial-targeting therapies
  • Drug safety – to investigate toxicology by looking at mitochondrial responses
  • Therapeutic delivery – monitoring the uptake of fluorescent drugs, nanoparticles or therapeutic mitochondria
  • Mitochondrial disease – to characterise mitochondrial dysfunction by studying fragmentation, swelling and network disruption

To learn more about how Nanolive’s Smart Mictochondrial AssayLIVE system works and what it can do, watch the on-demand webinar.