Imaging: Calibration Process
Fluorescence detection has generally not been considered a quantitative method for imaging applications. This is not without reason as light sources, optical filters, and a wide variety of other instabilities can results in varying image intensities. Further, the quantitative aspects of antibody detection are seldom characterized since they can be influenced by a number of factors such as access, sensitivity and specificity of the antibody, and microenvironment of the antigen. These and other issues have been gradually resolved through a number of methods developed over several years. (Full List) A summary of the important steps taken to ensure our measurements are quantifiable is given below.