This paper, for the first time, reports a method that can be used as a highly sensitive probe for singlet oxygen (1O2) and superoxide anion (O2-) in vitro or in vivo. FCLA(3,7- dihydro-6-{4-[2-(N′-(5-fluoresceinyl)thioureido)ethoxy]phenyl}-2-methylimidazo{1,2- a}pyrazin-3-one sodium salt), a chemiluminescence (CL) analysis reagent, has been reported to sensitively react with 1O2 and O2- to emit photons with a spectral peak of 525nm. In this work, when human serum albumin (HSA) was added into FCLA solution to enhance the CL intensity, approximately 20 times, compared to that without HSA. The enhanced CL had the same 525 nm spectral peak, identical to that without HSA. By gradually reducing the molecular oxygen content in the solution, we find that the auto-oxidation of oxygen molecules dissolved in the solution plays an important role in the CL process. Based on these experimental evidences, we propose a novel and highly sensitive detection method of 1O2 and O2-, which may have a great potential in chemical and medical applications.