In contrast, a chemiluminescent molecule is autonomously luminous. No excitation light is required for chemiluminescence, however the luminescence peaks within minutes of substrate addition and decays over just a few hours. Limits of detection. A chemiluminescence system that already uses alkaline phosphatase can become a chemifluorescent system simply by switching to the chemifluorescent substrate. Standard buffers and incubations remain the same.
Chemiluminescence is linear over about a tenfold range.
Limits of detection of chemifluorescence and chemiluminescence. Comparison of chemifluorescence vs. chemiluminescence protocols. Most chemiluminescence protocols recommend a short substrate incubation, prior to variable film exposure times to control background levels. Chemifluorescent signal does not decay rapidly, and Molecular Dynamics fluorescence imaging systems can be adjusted to optimize signal over background. Linear range of chemifluorescence vs. chemiluminescence.