Determining the Intracellular Organization of Organelles
Many studies have found alterations in the positioning and morphology of intracellular organelles under different experimental conditions. Although the precise quantification of these changes is challenging, it is strongly facilitated in single cells that are seeded on micropatterned substrates. Indeed, the controlled microenvironment of the cell leads to a reproducible distribution of organelles, simplifying image analysis and minimizing the number of cells required for robust phenotypes. Here, we outline how alterations in the intracellular organization of lysosomes and mitochondria, as a result of different growth conditions, can be efficiently quantified in cells seeded on adhesive micropatterns.
Key wordsMicropatterning Lysosome positioning Mitochondria morphology Organelle positioning Cell architecture
The authors greatly acknowledge the Cell and Tissue Imaging Facility (PICT-IBiSA @Burg and @Pasteur) and Nikon Imaging Center, Institut Curie (Paris), member of the French National Research Infrastructure France-BioImaging (ANR10-INBS-04). We thank Tarn Duong for advices on statistical analysis and kernel density estimation and Jean Philippe Grossier for providing scripts. This project was supported by grants from INFECT-ERA (ANR-14-IFEC-0002-04), the Centre National de la Recherche Scientifique and Institut Curie. The authors declare no conflict of interest.
- 14.Rasband W ImageJ. U. S. National Institutes of Health, Bethesda, MD, pp 1997–2016 https://imagej.nih.gov/ij/
- 19.Schauer K, Duong T, Gomes-Santos CS, Goud B (2014) Chapter 20 - Studying Intracellular trafficking pathways with probabilistic density maps. In: Perez F, Stephens DJ (eds) Methods in cell biology, vol 118. Academic Press, San Diego, CA, pp 325–343Google Scholar