New tools and approaches of quantum optics offer a unique opportunity to generate light pulses carrying a precise number of photons. Accurate control over the light pulses helps to improve the characterization of photo-induced processes. Here, we interface a specialized light source which provides flashes containing just one photon, with retinal rod cells of Xenopus laevis toads. We provide unambiguous proof of single photon sensitivity of rod cells without relying on the statistical modeling. We determined their quantum efficiencies without the use of any pre-calibrated detectors, and obtained the value of (29±4.7)%. Our approach provides the path for future studies and applications of quantum properties of light in phototransduction, vision, and photosynthesis.Ability to control light at a quantum level can be extremely useful in addressing biological problems. Interfacing biological objects with non-classical (quantum) light allows to enhance the precision of biological measurements [1], fosters development of more precise models of biological processes [2], and allows to reveal possible role of quantum effects in neurobiology [3,4] and perception [5,6].