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Determining the Intracellular Organization of Organelles

  • Bruno Latgé
  • Kristine Schauer
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1862)

Abstract

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 words

Micropatterning Lysosome positioning Mitochondria morphology Organelle positioning Cell architecture 

Notes

Acknowledgments

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.

Supplementary material

451006_1_En_19_MOESM1_ESM.txt (2 kb)
Annex 1 Macro1: Find Center of Micropatterns (TXT 1 kb)
451006_1_En_19_MOESM2_ESM.txt (2 kb)
Annex 2 Macro2: Batch Mode Segmentation Using 3D Object Counter (TXT 1 kb)
451006_1_En_19_MOESM3_ESM.txt (7 kb)
Annex 3 R Source File for KDE Computation (TXT 6 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Molecular Mechanisms of Intracellular Transport Group, Institut CuriePSL Research UniversityParis Cedex 05France
  2. 2.Centre National de la Recherche ScientifiqueUnité Mixte de Recherche 144ParisFrance

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