Subcellular Euclidean distance measurements with multicolor fluorescence localization imaging in cultured cells

Summary: This protocol measures the 3D Euclidean distance (Δ3D) between two/three fluorescently labeled kinetochore components in fixed samples using Kinetochore Delta software (KiDv1.0.1, MATLAB based). Overestimation of mean Δ3D is corrected through a Bayesian algorithm, with ΔEC distances reflect...

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Autores principales:	Tsvetelina E. Germanova, Emanuele Roscioli, Jonathan U. Harrison, Andrew D. McAinsh, Nigel J. Burroughs
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Cell Biology Cell culture Microscopy Science (General) Q1-390
Acceso en línea:	https://doaj.org/article/7c410bd5af754cd2836e928d7012fddf
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spelling	oai:doaj.org-article:7c410bd5af754cd2836e928d7012fddf2021-11-18T04:51:37ZSubcellular Euclidean distance measurements with multicolor fluorescence localization imaging in cultured cells2666-166710.1016/j.xpro.2021.100774https://doaj.org/article/7c410bd5af754cd2836e928d7012fddf2021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2666166721004809https://doaj.org/toc/2666-1667Summary: This protocol measures the 3D Euclidean distance (Δ3D) between two/three fluorescently labeled kinetochore components in fixed samples using Kinetochore Delta software (KiDv1.0.1, MATLAB based). Overestimation of mean Δ3D is corrected through a Bayesian algorithm, with ΔEC distances reflecting the ensemble average positions of fluorophores within a kinetochore population. This package also enables kinetochore categorization, which can be used to sub-sample kinetochores and measure ΔEC. Together, this allows the dynamic architecture of human kinetochores to be investigated (tested in hTERT-RPE1 cells).For complete details on the use and execution of this protocol, please refer to Roscioli et al. (2020).Tsvetelina E. GermanovaEmanuele RoscioliJonathan U. HarrisonAndrew D. McAinshNigel J. BurroughsElsevierarticleCell BiologyCell cultureMicroscopyScience (General)Q1-390ENSTAR Protocols, Vol 2, Iss 4, Pp 100774- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Cell Biology Cell culture Microscopy Science (General) Q1-390
spellingShingle	Cell Biology Cell culture Microscopy Science (General) Q1-390 Tsvetelina E. Germanova Emanuele Roscioli Jonathan U. Harrison Andrew D. McAinsh Nigel J. Burroughs Subcellular Euclidean distance measurements with multicolor fluorescence localization imaging in cultured cells
description	Summary: This protocol measures the 3D Euclidean distance (Δ3D) between two/three fluorescently labeled kinetochore components in fixed samples using Kinetochore Delta software (KiDv1.0.1, MATLAB based). Overestimation of mean Δ3D is corrected through a Bayesian algorithm, with ΔEC distances reflecting the ensemble average positions of fluorophores within a kinetochore population. This package also enables kinetochore categorization, which can be used to sub-sample kinetochores and measure ΔEC. Together, this allows the dynamic architecture of human kinetochores to be investigated (tested in hTERT-RPE1 cells).For complete details on the use and execution of this protocol, please refer to Roscioli et al. (2020).
format	article
author	Tsvetelina E. Germanova Emanuele Roscioli Jonathan U. Harrison Andrew D. McAinsh Nigel J. Burroughs
author_facet	Tsvetelina E. Germanova Emanuele Roscioli Jonathan U. Harrison Andrew D. McAinsh Nigel J. Burroughs
author_sort	Tsvetelina E. Germanova
title	Subcellular Euclidean distance measurements with multicolor fluorescence localization imaging in cultured cells
title_short	Subcellular Euclidean distance measurements with multicolor fluorescence localization imaging in cultured cells
title_full	Subcellular Euclidean distance measurements with multicolor fluorescence localization imaging in cultured cells
title_fullStr	Subcellular Euclidean distance measurements with multicolor fluorescence localization imaging in cultured cells
title_full_unstemmed	Subcellular Euclidean distance measurements with multicolor fluorescence localization imaging in cultured cells
title_sort	subcellular euclidean distance measurements with multicolor fluorescence localization imaging in cultured cells
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/7c410bd5af754cd2836e928d7012fddf
work_keys_str_mv	AT tsvetelinaegermanova subcellulareuclideandistancemeasurementswithmulticolorfluorescencelocalizationimaginginculturedcells AT emanueleroscioli subcellulareuclideandistancemeasurementswithmulticolorfluorescencelocalizationimaginginculturedcells AT jonathanuharrison subcellulareuclideandistancemeasurementswithmulticolorfluorescencelocalizationimaginginculturedcells AT andrewdmcainsh subcellulareuclideandistancemeasurementswithmulticolorfluorescencelocalizationimaginginculturedcells AT nigeljburroughs subcellulareuclideandistancemeasurementswithmulticolorfluorescencelocalizationimaginginculturedcells
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Subcellular Euclidean distance measurements with multicolor fluorescence localization imaging in cultured cells

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