The number of receptors opening after glutamate release is critical for understanding the sources of noise and the dynamic range of synaptic transmission. We imaged [Ca2+] transients mediated by synaptically activated NMDA receptors (NMDA-Rs) in individual spines in rat brain slices. We show that Ca2+ influx through single NMDA-Rs can be reliably detected, allowing us to estimate the number of receptors opening after synaptic transmission. This number is small: at the peak of the synaptic response, less than one NMDA-R is open, on average. Therefore, stochastic interactions between transmitter and receptor contribute substantially to synaptic noise, and glutamate occupies a small fraction of receptors. The number of receptors opening did not scale with spine volume, and smaller spines experience larger [Ca2+] transients during synaptic transmission. Our measurements further demonstrate that optical recordings can be used to study single receptors in intact systems
