Neuron-specific enolase and S100 protein are markers of neuronal lysis. To assess the neuronal suffering in rhegmatogenous retinal detachment we quantified neuron-specific enolase and S100 protein in the subretinal fluid. The puncture was performed in the sclera with a Merseture 5/0 round needle, and the fluid was collected with a glass capillary tube. Twelve subretinal fluid samples were obtained from 12 eyes with rhegmatogenous retinal detachment undergoing retinal detachment surgery. Vitreous from ten eyes with macular hole or epimacular membrane served as negative control group, and vitreous collected during cornea procurement from ten deceased patients served as positive control group. The mean concentration of neuron-specific enolase (in nanogrammes per millilitre) was 602 in the subretinal fluid of rhegmatogenous retinal detachment, 10.2 in the serum of these patients, 2.9 in the vitreous of the negative control group, and 364 in the positive control group. The mean concentration of S100 protein (in nanogrammes per millilitre) was 104 in the subretinal fluid of rhegmatogenous retinal detachment, <0.1 in the serum of these patients and in the vitreous of the control negative group, and 11.18 in the positive control group. Neuron-specific enolase (NSE) and S100 are known to be good markers of brain stress and, thus, are good markers of retinal stress.