The putative role of singlet oxygen (1O2) in the respiratory burst of neutrophils has remained elusive due to the lack of reliable means to study its quantitative production. To measure 1O2 directly from biological or chemical reactions in the near infrared region, we have developed a highly sensitive detection system which employs two InGaAs/InP pin photodiodes incorporated with a dual charge integrating amplifier circuit. Using this detection system, we detected light emission derived from a myeloperoxidase (MPO)-mediated reaction in physiological conditions: pH 7.4, 1-30 nM MPO, 10-100 microM H2O2 and 100-130 mM CI in place of Br without the use of deuterium oxide. The MNPO-H2O2-CI(-) system exhibited a single emission peak at 1.27 microm with a spectral distribution identical to that of delta singlet oxygen. Our results suggest physiological production of 1O2 in the MPO-H2O2-CI(-) system at an intravacuolar neutral pH. The MPO-mediated generation of 1O2, which may have an important role in host defense mechanisms, is discussed in connection with previous results.