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Flow Cytometry Protocols for Surface and Intracellular Antigen Analyses of Neural Cell Types
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1Emmy Noether-Group for Stem Cell Biology, Department of Molecular Embryology, Institute of Anatomy and Cell Biology,University of Freiburg, 2Spemann Graduate School of Biology and Medicine and Faculty of Biology,University of Freiburg, 3School of Life Sciences,Keele University, 4Center for Biological Signaling Studies (BIOSS),University of Freiburg
Capítulos
- 00:05Título
- 01:03Neural Cell Harvesting
- 03:10Antibody-independent Intracellular Labeling with Carboxyfluorescein Succinimidyl Ester (CFSE)
- 04:49Cell-surface Antigen Staining with Fluorophore-conjugated Antibodies
- 05:56Fixation and Membrane Permeabilization
- 07:35Intracellular Antigen Staining with Non-fluorescent Primary Antibodies
- 09:30Flow Cytometry Analysis
- 10:37Results: Multicolor Flow Cytometric Analysis of Surface vs. Introcellular Antigens in Neural Cells
- 12:36Conclusion
We provide a detailed description of a protocol for flow cytometric analysis of surface antigens and/or intracellular antigens in neural cell types. Critical aspects of experimental planning, step-by-step methodological procedures, and fundamental principles of flow cytometry are explained in order to enable neurobiologists to exploit this powerful technology.
