Responses of retinal rods to single photons

1. A suction electrode was used to record the membrane current of single rod outer segments in pieces of toad retina. During dim illumination the membrane current showed pronounced fluctuations. 2. Amplitude histograms of responses to dim flashes of fixed intensity exhibited two discrete peaks, one at 0 pA and one near 1 pA, suggesting that the response was quantized. By setting a criterion amplitude level, flash responses could be classed as 'failures' (no response) or as 'successes' (at least one quantal event). 3. The variation of fraction of successes with flash intensity was consistent with the hypothesis that each quantal electrical event resulted from a single photoisomerization. 4. The quantal event had a mean amplitude of about 1 pA (5% of the standing dark current) and a standard deviation of 0.2 pA. Dispersion in the event amplitude prevented identification of histogram peaks corresponding to two or more photoisomerizations. 5. Individual quantal responses exhibited a smooth shape very similar to that of the average quantal response. This suggests that a single photoisomerization releases many particles of transmitter and that radial diffusion of internal transmitter is not a major source of delay in the light response. 6. The 'quantum efficiency' with which an absorbed photon generated an electrical event was measured as 0.5 +/- 0.1 (S.E. of mean, n = 4). This is slightly lower than the quantum efficiency of photoisomerization obtained previously for rhodopsin in solution. 7. At wavelengths between 420 and 700 nm the quantal event was invariant in size, although the cell's sensitivity varied over a range of 10(5). 8. The power spectrum of the fluctuations in dim steady light was predicted by assuming that a random series of quantal events occurred independently. 9. In brighter light the fluctuations were faster, and the response to an incremental flash was reduced in size and duration. The power spectrum could be predicted by assuming random superposition of events with the shape of the incremental flash response.