title: Quantitative analysis of dynamic association in live biological fluorescent samples.
abstract: Determining vesicle localization and association in live microscopy may be challenging due to non-simultaneous imaging of rapidly moving objects with two excitation channels. Besides errors due to movement of objects, imaging may also introduce shifting between the image channels, and traditional colocalization methods cannot handle such situations. Our approach to quantifying the association between tagged proteins is to use an object-based method where the exact match of object locations is not assumed. Point-pattern matching provides a measure of correspondence between two point-sets under various changes between the sets. Thus, it can be used for robust quantitative analysis of vesicle association between image channels. Results for a large set of synthetic images shows that the novel association method based on point-pattern matching demonstrates robust capability to detect association of closely located vesicles in live cell-microscopy where traditional colocalization methods fail to produce results. In addition, the method outperforms compared Iterated Closest Points registration method. Results for fixed and live experimental data shows the association method to perform comparably to traditional methods in colocalization studies for fixed cells and to perform favorably in association studies for live cells.
authors.affiliation: Department of Signal Processing, Tampere University of Technology, Tampere, Finland; Department of Biological and Environmental Science/Nanoscience Center, University of Jyväskylä, Jyväskylä, Finland.
authors.forename: Pekka
authors.initials: P
authors.lastname: Ruusuvuori
authors.affiliation: Department of Biological and Environmental Science/Nanoscience Center, University of Jyväskylä, Jyväskylä, Finland; Department of Mathematical Information Technology, University of Jyväskylä, Jyväskylä, Finland.
authors.forename: Lassi
authors.initials: L
authors.lastname: Paavolainen
authors.affiliation: Department of Signal Processing, Tampere University of Technology, Tampere, Finland; Department of Mathematics, Tampere University of Technology, Tampere, Finland.
authors.forename: Kalle
authors.initials: K
authors.lastname: Rutanen
authors.affiliation: Department of Biological and Environmental Science/Nanoscience Center, University of Jyväskylä, Jyväskylä, Finland.
authors.forename: Anita
authors.initials: A
authors.lastname: Mäki
authors.affiliation: Department of Signal Processing, Tampere University of Technology, Tampere, Finland.
authors.forename: Heikki
authors.initials: H
authors.lastname: Huttunen
authors.affiliation: Department of Biological and Environmental Science/Nanoscience Center, University of Jyväskylä, Jyväskylä, Finland.
authors.forename: Varpu
authors.initials: V
authors.lastname: Marjomäki
mesh_terms: D045744:Cell Line, Tumor; D002470:Cell Survival; D003198:Computer Simulation; D022162:Cytoplasmic Vesicles / Q000378:metabolism*; D005453:Fluorescence; D006801:Humans; D008853:Microscopy; D057054:Molecular Imaging / Q000379:methods*; D059008:Time-Lapse Imaging
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