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Quantitative imaging of B1 cyclin expression across the cell cycle using green fluorescent protein tagging and epi-fluorescence

Karuna, Arnica, Masia, Francesco ORCID: https://orcid.org/0000-0003-4958-410X, Chappell, Sally, Errington, Rachel ORCID: https://orcid.org/0000-0002-8016-4376, Hartley, Andrew M., Jones, D. Dafydd ORCID: https://orcid.org/0000-0001-7709-3995, Borri, Paola ORCID: https://orcid.org/0000-0002-7873-3314 and Langbein, Wolfgang ORCID: https://orcid.org/0000-0001-9786-1023 2020. Quantitative imaging of B1 cyclin expression across the cell cycle using green fluorescent protein tagging and epi-fluorescence. Cytometry Part A 97 (10) , pp. 1066-1072. 10.1002/cyto.a.24038

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Abstract

In this work we report the number of cyclin B1 proteins tagged with enhanced green fluorescent protein (eGFP) in fixed U-2 OS cells across the cell cycle. We use a quantitative analysis of epi-fluorescence to determine the number of eGFP molecules in a non-destructive way, and integrated over the cell we find 104 to 105 molecules. Based on the measured number of eGFP tagged cyclin B1 proteins, knowledge of cyclin B1 dynamics through the cell cycle, and the cell morphology, we identify the stages of cells in the cell cycle.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Biosciences
Medicine
Physics and Astronomy
Publisher: Wiley
ISSN: 1552-4922
Funders: EPSRC, BBSRC
Date of First Compliant Deposit: 28 April 2020
Date of Acceptance: 28 April 2020
Last Modified: 11 Oct 2023 18:17
URI: https://orca.cardiff.ac.uk/id/eprint/131272

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