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Subcellular Euclidean distance measurements with multicolor fluorescence localization imaging in cultured cells
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Germanova, Tsvetelina E., Roscioli, Emanuele, Harrison, Jonathan, McAinsh, Andrew D. and Burroughs, Nigel John (2021) Subcellular Euclidean distance measurements with multicolor fluorescence localization imaging in cultured cells. STAR Protocols, 2 (4). 100774. doi:10.1016/j.xpro.2021.100774 ISSN 2666-1667.
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Official URL: https://doi.org/10.1016/j.xpro.2021.100774
Abstract
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).
Item Type: | Journal Article | |||||||||||||||
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Subjects: | Q Science > QA Mathematics Q Science > QH Natural history |
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Mathematics Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School |
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SWORD Depositor: | Library Publications Router | |||||||||||||||
Library of Congress Subject Headings (LCSH): | Fluorescence microscopy -- Methodology, Microscopy -- Methodology, Distance geometry, Euclidean algorithm | |||||||||||||||
Journal or Publication Title: | STAR Protocols | |||||||||||||||
Publisher: | Elsevier BV | |||||||||||||||
ISSN: | 2666-1667 | |||||||||||||||
Official Date: | 17 December 2021 | |||||||||||||||
Dates: |
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Volume: | 2 | |||||||||||||||
Number: | 4 | |||||||||||||||
Article Number: | 100774 | |||||||||||||||
DOI: | 10.1016/j.xpro.2021.100774 | |||||||||||||||
Status: | Peer Reviewed | |||||||||||||||
Publication Status: | Published | |||||||||||||||
Access rights to Published version: | Open Access (Creative Commons) | |||||||||||||||
Date of first compliant deposit: | 26 November 2021 | |||||||||||||||
Date of first compliant Open Access: | 29 November 2021 | |||||||||||||||
RIOXX Funder/Project Grant: |
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