ContactJ: Characterization of lipid droplet-mitochondrial contacts using fluorescence microscopy and image analysis.
Maria CalvoMarta BoschElisenda CollRobert G PartonAlbert PolPublished in: F1000Research (2021)
Lipid droplets (LDs) are the major lipid storage organelles of eukaryotic cells and together with mitochondria key regulators of cell bioenergetics. LDs communicate with mitochondria and other organelles forming "metabolic synapse" contacts to ensure that lipid supply occurs where and when necessary. Although transmission electron microscopy analysis allows an accurate and precise analysis of contacts, the characterization of a large number of cells and conditions can become a long-term process. In order to extend contact analysis to hundreds of cells and multiple conditions, we have combined confocal fluorescence microscopy with advanced image analysis methods. In this work, we have developed the ImageJ macro script ContactJ, a novel and straight image analysis method to identify and quantify contacts between LD and mitochondria in fluorescence microscopy images allowing the automatic analysis. This image analysis workflow combines colocalization and skeletonization methods, enabling the quantification of LD-mitochondria contacts together with a complete characterization of organelles and cellular parameters. The correlation and normalization of these parameters contribute to the complex description of cell behavior under different experimental energetic states. ContactJ is available here: https://doi.org/10.5281/zenodo.5810874.
Keyphrases
- single molecule
- induced apoptosis
- cell cycle arrest
- cell death
- optical coherence tomography
- high resolution
- single cell
- high throughput
- reactive oxygen species
- fatty acid
- oxidative stress
- deep learning
- endoplasmic reticulum
- stem cells
- signaling pathway
- cell therapy
- endoplasmic reticulum stress
- mesenchymal stem cells
- transcription factor
- cell proliferation
- energy transfer
- quantum dots
- pi k akt