The collection of time-series of periphyton biomass is a difficult task due to the destructive nature of the standard methods. A non-destructive method based on photography and digitalization, for the estimation of Chla of periphyton colonizing artificial substrata is presented. The standard spectrophotometric method was used to obtain a calibration curve. The relative errors of the proposed method were similar to those of other published methods. The photographic method should be used when a large quantity of samples from the same community is needed and a high precision on the individual measurement is not required.