Superoxide Production in the Islet of Langerhans Detected by the MCLA Chemiluminescence Method

It is well known that the superoxide anion radical (O-
2
) is produced in the body as a result of biochemical activities of many cells, such as neutrophils, macrophages, and Kupffer cells (
1
). Production of O-
2
is detectable by various methods, including cytochrome-c oxidation (
1
), nitroblue tetrazolium (NBT) staining, and chemiluminescence (CL) measurement (
2
). The site and amount of O-
2
production is variable with time and readily determined in vivo by applying the CL method. In earlier studies, the CL method was successfully applied for detection of O-
2
generation at a tissue level in the lung (
3
), liver (
4
), pancreas (
5
), heart (
6
), and brain (
7
). Combined with video imaging techniques, the CL method makes it possible to analyze O-
2
generation at a high spatial resolution, sometimes even at a single cell level. In this chapter, we describe the CL method applied to O-
2
detection from the islets of Langerhans in the perfused pancreas.