Instrumentation methods currently used in endodontic therapy create a residual layer composed of organic and inorganic material that frequently retains bacteria and their subproducts. The available methods to remove this smear layer include chemical etching, ultrasonic and laser techniques but none of these is completely efficient and there is no universal consensus regarding this subject. In the present study the effect of EDTA, EDTAC and citric acid upon radicular dentin was analyzed by two methods. Dentin microhardness was measured before and after etching. Atomic force microscopy was used to map and characterize dentinary microstructural changes throughout the chelation process. Real time, in situ, observations of the process were performed through the use of a special sample holder - a liquid cell - that allowed the observation of the samples surfaces during etching. Image sequences were acquired and an image processing and analysis procedure was developed to evaluate, both qualitatively and quantitatively, the effect of the 3 chelators. The development of this methodology employing in situ microscopy and digital image analysis is one of the main contributions of the present dissertation.