- Quantitative Imaging of B1 Cyclin Expression Across the Cell Cycle Using Green Fluorescent Protein Tagging and Epifluorescence.
Quantitative Imaging of B1 Cyclin Expression Across the Cell Cycle Using Green Fluorescent Protein Tagging and Epifluorescence.
Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020-07-03)
Arnica Karuna, Francesco Masia, Sally Chappell, Rachel Errington, Andrew M Hartley, Darran Dafydd Jones, Paola Borri, Wolfgang Langbein
PMID32613720
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 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. © 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC. on behalf of International Society for Advancement of Cytometry.
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Cell Dissociation Solution Non-enzymatic 1x, Prepared in phosphate buffered saline without calcium and magnesium, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Penicillin-Streptomycin, with 10,000 units penicillin and 10 mg streptomycin per mL in 0.9% NaCl, 0.1 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
G 418 disulfate salt solution, 50 mg/mL in H2O, 0.1 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
McCoy′s 5A Medium, Modified, with sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture