Quantitative Imaging of B1 Cyclin Expression Across the Cell Cycle Using Green Fluorescent Protein Tagging and Epifluorescence. Issue 10 (1st July 2020)

• Record Type: Journal Article
• Authors: Karuna, Arnica; Masia, Francesco; Chappell, Sally; Errington, Rachel; Hartley, Andrew M.; Jones, Darran Dafydd; Borri, Paola; Langbein, Wolfgang
• Is Part Of: Cytometry. Volume 97:Issue 10(2020)
• Page Start: 1066
• Page End: 1072
• Publication Date: 2020-07-01
• Subjects: fluorescence microscopy -- fluorophore concentration -- U‐2 OS cells -- cyclin B1‐eGFP; Flow cytometry -- Periodicals; Imaging systems in biology -- Periodicals; Imaging systems in medicine -- Periodicals; Diagnostic imaging -- Periodicals; 571.605
• DOI: 10.1002/cyto.a.24038
• Languages: English
• ISSNs: 1552-4922

Abstract

Abstract: In this article, 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 epifluorescence to determine the number of eGFP molecules in a nondestructive way, and integrated over the cell we find 10 4 to 10 5 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. © 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC. on behalf of International Society for Advancement of Cytometry. Abstract : eGFP tagged cyclin B1 is imaged using epifluorescence microscopy in U‐2 OS cells, and quantitatively calibrated. The number of molecules and concentration of eGFP are determined along the cell cycle.