title: Quantifying lymphocyte kinetics in vivo using carboxyfluorescein diacetate succinimidyl ester (CFSE).
abstract: The cytoplasmic dye carboxyfluorescein diacetate succinimidyl ester (CFSE) is used to quantify cell kinetics. It is particularly important in studies of lymphocyte homeostasis where its labelling of cells irrespective of their stage in the cell cycle makes it preferable to deuterated glucose and BrdU, which only label dividing cells and thus produce unrepresentative results. In the past, experiments have been limited by the need to obtain a clear separation of CFSE peaks forcing scientists to adopt a strategy of in vitro labelling of cells followed by their injection into the host. Here we develop a framework for analysis of in vivo CFSE labelling data. This enables us to estimate the rate of proliferation and death of lymphocytes in situ, and thus represents a considerable advance over current procedures. We illustrate this approach using in vivo CFSE labelling of B lymphocytes in sheep.
authors.affiliation: Department of Immunology, Imperial College London, Norfolk Place, London W2 1PG, UK. b.asquith@ic.ac.uk
authors.forename: Becca
authors.initials: B
authors.lastname: Asquith
authors.forename: Christophe
authors.initials: C
authors.lastname: Debacq
authors.forename: Arnaud
authors.initials: A
authors.lastname: Florins
authors.forename: Nicolas
authors.initials: N
authors.lastname: Gillet
authors.forename: Teresa
authors.initials: T
authors.lastname: Sanchez-Alcaraz
authors.forename: Angelina
authors.initials: A
authors.lastname: Mosley
authors.forename: Luc
authors.initials: L
authors.lastname: Willems
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism / Q000502:physiology*; D002455:Cell Division; D005434:Flow Cytometry; D005452:Fluoresceins / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism*; D007700:Kinetics; D008433:Mathematics; D012756:Sheep; D013388:Succinimides / Q000378:metabolism*
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D005452:Fluoresceins; D005456:Fluorescent Dyes; D013388:Succinimides