Inhomogeneity Based Characterization of Distribution Patterns on the Plasma Membrane.
Cell surface protein and lipid molecules are organized in various patterns: randomly, along gradients, or clustered when segregated into discrete micro- and nano-domains. Their distribution is tightly coupled to events such as polarization, endocytosis, and intracellular signaling, but challenging to quantify using traditional techniques. Here we present a novel approach to quantify the distribution of plasma membrane proteins and lipids. This approach describes spatial patterns in degrees of inhomogeneity and incorporates an intensity-based correction to analyze images with a wide range of resolutions; we have termed it Quantitative Analysis of the Spatial distributions in Images using Mosaic segmentation and Dual parameter Optimization in Histograms (QuASIMoDOH). We tested its applicability using simulated microscopy images and images acquired by widefield microscopy, total internal reflection microscopy, structured illumination microscopy, and photoactivated localization microscopy. We validated QuASIMoDOH, successfully quantifying the distribution of protein and lipid molecules detected with several labeling techniques, in different cell model systems. We also used this method to characterize the reorganization of cell surface lipids in response to disrupted endosomal trafficking and to detect dynamic changes in the global and local organization of epidermal growth factor receptors across the cell surface. Our findings demonstrate that QuASIMoDOH can be used to assess protein and lipid patterns, quantifying distribution changes and spatial reorganization at the cell surface. An ImageJ/Fiji plugin of this analysis tool is provided.
VIB Bio Imaging Core, Herestraat, Leuven, Belgium.|Laboratory of Membrane Trafficking, Department of Human Genetics, KU Leuven, Herestraat, Leuven, Belgium.|VIB Center for the Biology of Disease, KU Leuven, Herestraat, Leuven, Belgium.
Laura
L
Paparelli
VIB Bio Imaging Core, Herestraat, Leuven, Belgium.|VIB Center for the Biology of Disease, KU Leuven, Herestraat, Leuven, Belgium.|VIB, LiMoNe, Herestraat, Leuven, Belgium.
Nikky
N
Corthout
VIB Bio Imaging Core, Herestraat, Leuven, Belgium.|VIB Center for the Biology of Disease, KU Leuven, Herestraat, Leuven, Belgium.
Benjamin
0000-0002-0249-3844
B
Pavie
Department of Molecular Medicine, Beckman Research Institute of the City of Hope Comprehensive Cancer Center, Duarte, California, United States of America.
Devin L
DL
Wakefield
Laboratory of Membrane Trafficking, Department of Human Genetics, KU Leuven, Herestraat, Leuven, Belgium.|VIB Center for the Biology of Disease, KU Leuven, Herestraat, Leuven, Belgium.
Ragna
R
Sannerud
Department of Molecular Medicine, Beckman Research Institute of the City of Hope Comprehensive Cancer Center, Duarte, California, United States of America.
Tijana
T
Jovanovic-Talisman
Laboratory of Membrane Trafficking, Department of Human Genetics, KU Leuven, Herestraat, Leuven, Belgium.|VIB Center for the Biology of Disease, KU Leuven, Herestraat, Leuven, Belgium.
Wim
W
Annaert
VIB Bio Imaging Core, Herestraat, Leuven, Belgium.|VIB Center for the Biology of Disease, KU Leuven, Herestraat, Leuven, Belgium.|VIB, LiMoNe, Herestraat, Leuven, Belgium.
Sebastian
S
Munck
D000465:Algorithms; D000818:Animals; D002462:Cell Membrane / Q000737:chemistry*; D002478:Cells, Cultured; D019295:Computational Biology / Q000379:methods*; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008055:Lipids / Q000032:analysis* / Q000737:chemistry; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry; D051379:Mice; D008856:Microscopy, Fluorescence; D012984:Software
D008055:Lipids; D008565:Membrane Proteins