第二章 SP定量观察法.pdfVIP

q 2007 International Society for Analytical Cytology Cytometry Part A 71A:251–257 (2007) Combination of Quantification and Observation Methods for Study of ‘‘Side Population’’ Cells in Their ‘‘In Vitro’’ Microenvironment Rachid Benchaouir,1 Julien Picot, 1 Nicolas Greppo,1 Philippe Rameau,1 Daniel Stockholm,1 1 1,2 1* Luis Garcia, Andras Paldi, and Corinne Laplace-Builhe 1GENETHON Centre National de la Recherche Scientifique UMR 8115, 1 bis, rue de l’Internationale 91002 Evry, France 2Ecole Pratique des Hautes Etudes, 46 rue de Lille, 75007 Paris, France Received 15 June 2006; Revision received 27 October 2006; Accepted 28 November 2006 Background: Qualitative and quantitative analyses of in the plate. The cells with a given fluorescence intensity the rare phenotypic variants in in vitro culture systems is can then be located easily on the recorded image of the necessary for the understanding of cell differentiation in culture plate for further analysis. cell culture of primary cells or cell lines. Slide-based cyto- Results: The potential of our method is illustrated by the metry combines image acquisition and data treatment, identification and localization of SP cells in the cultures of and associates the power of flow cytometry (FCM) and the C2C12 cell line. Although these cells represent only the resolution of the microscopic studies making it suita- about 1% of the