Overview
This protocol describes a method to assess concentrations of free cytoplasmic calcium, [Ca2+]i, for cultured adherent RAW 264.7 cells in an 8-well coverglass. This objective is accomplished using the Ca2+-sensitive fluorescent dye, fura-2 acetoxymethyl (AM), which permeates cell membranes as an ester and is hydrolyzed in the cell to its Ca2+-sensitive acidic form. Fluorescence for the adherent cells is measured over time with cells that have been washed free of extracellular dye. Free [Ca2+] is estimated as described below (see Image and Data Processing).
Details
Details of this protocol, Live Single-Cell Fura-2 Measurements To Determine The Intracellular Free Calcium In RAW 264.7 Cells (with Olympus IX81), are located on a web site other than Biocompare Protocols.