Biology Reference
In-Depth Information
CHAPTER
10
Fluorescence Correlation
Spectroscopy and
Photon-Counting
Histogram Analysis
of Receptor-Receptor
Interactions
Katharine Herrick-Davis and Joseph E. Mazurkiewicz
Center for Neuropharmacology & Neuroscience, Albany Medical College, Albany, New York, USA
CHAPTER OUTLINE
Introduction ............................................................................................................ 182
10.1 Materials........................................................................................................184
10.2 Methods .........................................................................................................185
10.2.1 Sample Preparation .................................................................... 185
10.2.1.1 Choice of Cell Type.............................................................
185
10.2.1.2 Choice of Fluorescent Tag...................................................
185
10.2.1.3 Labeling the Receptor .........................................................
185
10.2.1.4 Plating Cells .......................................................................186
10.2.1.5 Transfecting Cells ...............................................................186
10.2.2 Instrument Setup ....................................................................... 186
10.2.2.1 Environment .......................................................................186
10.2.2.2 Imaging Setup ....................................................................186
10.2.2.3 FCS Setup ..........................................................................186
10.2.2.4 Instrument Alignment and Calibration .................................187
10.2.3 Data Acquisition......................................................................... 188
10.2.3.1 FCS Recording ...................................................................188
10.2.4 Data Analysis ............................................................................. 190
10.2.4.1 Diffusion Coefficient ............................................................190
10.2.4.2 Molecular Brightness ..........................................................192
10.3 Discussion......................................................................................................194
Summary ................................................................................................................ 195
Acknowledgments ................................................................................................... 195
References ............................................................................................................. 195
