Image Correlation Spectroscopy to Define Membrane Dynamics

Fluorescent imaging techniques are powerful tools that aid in studying protein dynamics and membrane domains and allow for the visualization and data collection of such structures as caveolae and clathrin-coated pits, key players in the regulation of cell communication and signaling. The family of image correlation spectroscopy (FICS) provides a unique way to determine details about aggregation, clustering, and dynamics of proteins on the plasma membrane. FICS consists of many imaging techniques which we will focus on including image correlation spectroscopy, image cross-correlation spectroscopy and dynamic image correlation spectroscopy. Image correlation spectroscopy is a tool used to calculate the cluster density, which is the average number of clusters per unit area along with data to determine the degree of aggregation of plasma membrane proteins. Image cross-correlation spectroscopy measures the colocalization of proteins of interest. Dynamic image correlation spectroscopy can be used to analyze protein aggregate dynamics on the cell surface during live-cell imaging in the millisecond to second range.