Modern image detectors with exceptionally low readout noise of about one electron (e-) per pixel allow for applications with ultra-low levels of light intensity. In this Letter, we report a property of scientific CMOS detectors that makes accurate spectroscopy at ultra-low levels of illumination depending on a thorough calibration procedure. Our results reveal that pixel sensitivity to light may have significant nonlinearity at accumulation levels smaller than 50e- per pixel. The sensitivity decreases by a factor of ∼0.7 at an accumulation level of ∼1e- per pixel and photon detection rate of about 170 Hz. We demonstrate that the nature of this nonlinearity might be quite complicated: the photoelectric response of a pixel depends on both the number of accumulated electrons and the detection count rate (at rates larger than 250 Hz).