DIGITAL QUANTIFICATION OF THE ENZYME-LINKED IMMUNOSPOT (ELISPOT)

Enzyme-linked immunospot (ELISPOT) is a sensitive technique for the de tection of cytokines released by immune cells. The technique is well e stablished and correlates closely with the enzyme-linked immunosorbent assay (ELISA) technique. Here, we introduce an integrated imaging-ana lysis system to quantitate the spots formed by the ELISPOT assay The s pots cart be easily counted when the background is clear and the spots are few. However; when the number of spots increases to >100, this ta sk becomes challenging find time-consuming. To minimize time and stand ardize the counting procedure, we have used a digital camera linked to a computer system for reading the plates. In general, the computer is able to count more spots and has a smaller standard deviation when co mpared with the manual microscopic count. This integrated system is co mmercially available, and we believe that this method is objective, ti mesaving and consistent in routine ELISPOT assay counting, especially when numerous spots are present.