The capability of acquiring hyperspectral information in low light conditions is potentially important for a variety of applications, ranging from remote sensing to biomedical fluorescence imaging. Particularly interesting is its use in optical analysis of biological samples in which the light level should be kept low to prevent tissue damage. For this purpose a low-light hyperspectral camera has been developed for the 0.4 to 0.9 µm spectral range. The camera is based on an electron-multiplying CCD (EMCCD) detector which effectively suppresses readout noise, and approaches the fundamental photon noise limit. It has been designed with close-up optics to image an area on the order of centimeters. Fluorescence images of skin samples illustrate the camera performance. Results show that low-light hyperspectral imaging has a potential for biomedical applications.