New algorithm to determine true colocalization in combination with image restoration and time-lapse confocal microscopy to map Kinases in mitochondria

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The subcellular localization and physiological functions of biomolecules are closely related and thus it is crucial to precisely determine the distribution of different molecules inside the intracellular structures. This is frequently accomplished by fluorescence microscopy with well-characterized m...

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Autores principales:	Villalta, J.I., Galli, S., Iacaruso, M.F., Arciuch, V.G.A., Poderoso, J.J., Jares-Erijman, E.A., Pietrasanta, L.I.
Formato:	JOUR
Materias:	mitogen activated protein kinase biological marker green fluorescent protein accuracy algorithm article automation cellular distribution confocal microscopy controlled study fluorescence imaging human human cell image analysis image reconstruction mitochondrion molecular imaging optical resolution protein localization reliability signal detection time lapse imaging animal cell strain 3T3 fluorescence microscopy HeLa cell image processing metabolism methodology mouse physiology plasmid signal transduction time Algorithms Animals Biological Markers Green Fluorescent Proteins HeLa Cells Humans Image Processing, Computer-Assisted MAP Kinase Signaling System Mice Microscopy, Confocal Microscopy, Fluorescence Mitochondria NIH 3T3 Cells Plasmids Time Factors
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_19326203_v6_n4_p_Villalta
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_19326203_v6_n4_p_Villalta
record_format	dspace
spelling	todo:paper_19326203_v6_n4_p_Villalta2023-10-03T16:35:09Z New algorithm to determine true colocalization in combination with image restoration and time-lapse confocal microscopy to map Kinases in mitochondria Villalta, J.I. Galli, S. Iacaruso, M.F. Arciuch, V.G.A. Poderoso, J.J. Jares-Erijman, E.A. Pietrasanta, L.I. mitogen activated protein kinase biological marker green fluorescent protein accuracy algorithm article automation cellular distribution confocal microscopy controlled study fluorescence imaging human human cell image analysis image reconstruction mitochondrion molecular imaging optical resolution protein localization reliability signal detection time lapse imaging algorithm animal cell strain 3T3 confocal microscopy fluorescence microscopy HeLa cell image processing metabolism methodology mouse physiology plasmid signal transduction time Algorithms Animals Biological Markers Green Fluorescent Proteins HeLa Cells Humans Image Processing, Computer-Assisted MAP Kinase Signaling System Mice Microscopy, Confocal Microscopy, Fluorescence Mitochondria NIH 3T3 Cells Plasmids Time Factors The subcellular localization and physiological functions of biomolecules are closely related and thus it is crucial to precisely determine the distribution of different molecules inside the intracellular structures. This is frequently accomplished by fluorescence microscopy with well-characterized markers and posterior evaluation of the signal colocalization. Rigorous study of colocalization requires statistical analysis of the data, albeit yet no single technique has been established as a standard method. Indeed, the few methods currently available are only accurate in images with particular characteristics. Here, we introduce a new algorithm to automatically obtain the true colocalization between images that is suitable for a wide variety of biological situations. To proceed, the algorithm contemplates the individual contribution of each pixel's fluorescence intensity in a pair of images to the overall Pearsońs correlation and Manders' overlap coefficients. The accuracy and reliability of the algorithm was validated on both simulated and real images that reflected the characteristics of a range of biological samples. We used this algorithm in combination with image restoration by deconvolution and time-lapse confocal microscopy to address the localization of MEK1 in the mitochondria of different cell lines. Appraising the previously described behavior of Akt1 corroborated the reliability of the combined use of these techniques. Together, the present work provides a novel statistical approach to accurately and reliably determine the colocalization in a variety of biological images. © 2011 Villalta et al. Fil:Galli, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Jares-Erijman, E.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_19326203_v6_n4_p_Villalta
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	mitogen activated protein kinase biological marker green fluorescent protein accuracy algorithm article automation cellular distribution confocal microscopy controlled study fluorescence imaging human human cell image analysis image reconstruction mitochondrion molecular imaging optical resolution protein localization reliability signal detection time lapse imaging algorithm animal cell strain 3T3 confocal microscopy fluorescence microscopy HeLa cell image processing metabolism methodology mouse physiology plasmid signal transduction time Algorithms Animals Biological Markers Green Fluorescent Proteins HeLa Cells Humans Image Processing, Computer-Assisted MAP Kinase Signaling System Mice Microscopy, Confocal Microscopy, Fluorescence Mitochondria NIH 3T3 Cells Plasmids Time Factors
spellingShingle	mitogen activated protein kinase biological marker green fluorescent protein accuracy algorithm article automation cellular distribution confocal microscopy controlled study fluorescence imaging human human cell image analysis image reconstruction mitochondrion molecular imaging optical resolution protein localization reliability signal detection time lapse imaging algorithm animal cell strain 3T3 confocal microscopy fluorescence microscopy HeLa cell image processing metabolism methodology mouse physiology plasmid signal transduction time Algorithms Animals Biological Markers Green Fluorescent Proteins HeLa Cells Humans Image Processing, Computer-Assisted MAP Kinase Signaling System Mice Microscopy, Confocal Microscopy, Fluorescence Mitochondria NIH 3T3 Cells Plasmids Time Factors Villalta, J.I. Galli, S. Iacaruso, M.F. Arciuch, V.G.A. Poderoso, J.J. Jares-Erijman, E.A. Pietrasanta, L.I. New algorithm to determine true colocalization in combination with image restoration and time-lapse confocal microscopy to map Kinases in mitochondria
topic_facet	mitogen activated protein kinase biological marker green fluorescent protein accuracy algorithm article automation cellular distribution confocal microscopy controlled study fluorescence imaging human human cell image analysis image reconstruction mitochondrion molecular imaging optical resolution protein localization reliability signal detection time lapse imaging algorithm animal cell strain 3T3 confocal microscopy fluorescence microscopy HeLa cell image processing metabolism methodology mouse physiology plasmid signal transduction time Algorithms Animals Biological Markers Green Fluorescent Proteins HeLa Cells Humans Image Processing, Computer-Assisted MAP Kinase Signaling System Mice Microscopy, Confocal Microscopy, Fluorescence Mitochondria NIH 3T3 Cells Plasmids Time Factors
description	The subcellular localization and physiological functions of biomolecules are closely related and thus it is crucial to precisely determine the distribution of different molecules inside the intracellular structures. This is frequently accomplished by fluorescence microscopy with well-characterized markers and posterior evaluation of the signal colocalization. Rigorous study of colocalization requires statistical analysis of the data, albeit yet no single technique has been established as a standard method. Indeed, the few methods currently available are only accurate in images with particular characteristics. Here, we introduce a new algorithm to automatically obtain the true colocalization between images that is suitable for a wide variety of biological situations. To proceed, the algorithm contemplates the individual contribution of each pixel's fluorescence intensity in a pair of images to the overall Pearsońs correlation and Manders' overlap coefficients. The accuracy and reliability of the algorithm was validated on both simulated and real images that reflected the characteristics of a range of biological samples. We used this algorithm in combination with image restoration by deconvolution and time-lapse confocal microscopy to address the localization of MEK1 in the mitochondria of different cell lines. Appraising the previously described behavior of Akt1 corroborated the reliability of the combined use of these techniques. Together, the present work provides a novel statistical approach to accurately and reliably determine the colocalization in a variety of biological images. © 2011 Villalta et al.
format	JOUR
author	Villalta, J.I. Galli, S. Iacaruso, M.F. Arciuch, V.G.A. Poderoso, J.J. Jares-Erijman, E.A. Pietrasanta, L.I.
author_facet	Villalta, J.I. Galli, S. Iacaruso, M.F. Arciuch, V.G.A. Poderoso, J.J. Jares-Erijman, E.A. Pietrasanta, L.I.
author_sort	Villalta, J.I.
title	New algorithm to determine true colocalization in combination with image restoration and time-lapse confocal microscopy to map Kinases in mitochondria
title_short	New algorithm to determine true colocalization in combination with image restoration and time-lapse confocal microscopy to map Kinases in mitochondria
title_full	New algorithm to determine true colocalization in combination with image restoration and time-lapse confocal microscopy to map Kinases in mitochondria
title_fullStr	New algorithm to determine true colocalization in combination with image restoration and time-lapse confocal microscopy to map Kinases in mitochondria
title_full_unstemmed	New algorithm to determine true colocalization in combination with image restoration and time-lapse confocal microscopy to map Kinases in mitochondria
title_sort	new algorithm to determine true colocalization in combination with image restoration and time-lapse confocal microscopy to map kinases in mitochondria
url	http://hdl.handle.net/20.500.12110/paper_19326203_v6_n4_p_Villalta
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New algorithm to determine true colocalization in combination with image restoration and time-lapse confocal microscopy to map Kinases in mitochondria

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