Cyclic guanosine 3′-5′-monophosphate (cGMP) plays a key role in the physiological process of light detection in photoreceptor cells of the retina. However, there is also growing evidence that cGMP may be critically involved in some pathophysiological processes of the retina since degenerating photoreceptors in mouse models of retinitis pigmentosa and achromatopsia accumulate high levels of cGMP. Here, we describe methods that allow the detection, subcellular localization, and quantification of cGMP in the retina and propose that cGMP accumulation can be used as a biomarker for photoreceptor degeneration.