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Quantitative Analysis of Dynamic Association in Live Biological Fluorescent Samples
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 co...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3984138/ https://www.ncbi.nlm.nih.gov/pubmed/24728133 http://dx.doi.org/10.1371/journal.pone.0094245 |
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author | Ruusuvuori, Pekka Paavolainen, Lassi Rutanen, Kalle Mäki, Anita Huttunen, Heikki Marjomäki, Varpu |
author_facet | Ruusuvuori, Pekka Paavolainen, Lassi Rutanen, Kalle Mäki, Anita Huttunen, Heikki Marjomäki, Varpu |
author_sort | Ruusuvuori, Pekka |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-3984138 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-39841382014-04-15 Quantitative Analysis of Dynamic Association in Live Biological Fluorescent Samples Ruusuvuori, Pekka Paavolainen, Lassi Rutanen, Kalle Mäki, Anita Huttunen, Heikki Marjomäki, Varpu PLoS One Research Article 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. Public Library of Science 2014-04-11 /pmc/articles/PMC3984138/ /pubmed/24728133 http://dx.doi.org/10.1371/journal.pone.0094245 Text en © 2014 Ruusuvuori et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Ruusuvuori, Pekka Paavolainen, Lassi Rutanen, Kalle Mäki, Anita Huttunen, Heikki Marjomäki, Varpu Quantitative Analysis of Dynamic Association in Live Biological Fluorescent Samples |
title | Quantitative Analysis of Dynamic Association in Live Biological Fluorescent Samples |
title_full | Quantitative Analysis of Dynamic Association in Live Biological Fluorescent Samples |
title_fullStr | Quantitative Analysis of Dynamic Association in Live Biological Fluorescent Samples |
title_full_unstemmed | Quantitative Analysis of Dynamic Association in Live Biological Fluorescent Samples |
title_short | Quantitative Analysis of Dynamic Association in Live Biological Fluorescent Samples |
title_sort | quantitative analysis of dynamic association in live biological fluorescent samples |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3984138/ https://www.ncbi.nlm.nih.gov/pubmed/24728133 http://dx.doi.org/10.1371/journal.pone.0094245 |
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