Fluorescence and Luminescence Studies of in Vivo Chlorophyll With a Laser Phosphoroscope*

A simple apparatus using a He‐Ne laser and a rotating sector gives repetitive light pulses with rise and fall times of a few μsec and intensities of up to 0.65 W/cm2. Fluorescence during the actinic pulse and luminescence following it (after 20 μsec) may be recorded and compared. Sampling of the suspension of photosynthetic cells or chloroplasts by intermittent flow permits the averaging of 25 such measurements. With Chlorella, the fast (30 μsec) and the medium (30 msec) components of luminescence do not saturate at the same light intensity. The former saturates in the same range as the photochemical fluorescence rise, whereas the latter saturates at higher intensities. Under a variety of conditions, the decay of luminescence intensity is always strongly polyphasic. Analysis of the kinetics discloses a dependency with the log concentration of the hypothetical precursor, suggesting a connection of luminescence with electrochemical phenomena. The shape of the luminescence integral, as well as the light saturation pattern, suggests another kind of interpretation, in terms of several pools, possibly two of equal size. In the presence of hydroxylamine, only one pool is seen. The stimulating effect of preillumination is only observed for the medium component. However, a stimulation of short duration also exists for the fast component. The problems of number of pools, mechanism, and stimulation are briefly discussed.