Gluma Dentin Bond is an adhesive system, where the primer contains 5% glutaraldehyde and 35% hydroxyethyl methacrylate. Practitioners have reported a strong desensitizing effect of the Gluma system on dentin. This study, thus, sought to evaluate the effect of this system on dentin using various microscopic techniques. 12 non-restored human molars extracted for prosthodontic reasons were used. Prior to extraction the buccal cusps were removed such that a 2 mm x 2 mm wide dentin surface was exposed. The surfaces were treated in 6 ways: (1) application of Gluma 2 cleanser, Gluma 3 primer to which 0.1% w/v fluorescein was added, and Gluma 4 sealer; (2) as in (1) but treatment with H2O/0.1% w/v fluorescein instead of the Gluma 3; (3) as in (1) but without Gluma 2; (4) as in (1) but with application of 5% glutaraldehyde instead of Gluma 3; (5) as in (1) but without Gluma 4; (6) as in (1) but with application of 35% HEMA/0.1% w/v fluorescein instead of Gluma 3. Following extraction, 1 tooth per procedure was prepared for confocal laser scanning microscopy. The remaining teeth were fixed and prepared for SEM and TEM evaluation. In specimens of procedures (1) and (5), tubular occlusions could be seen to a depth of 200 microm. In specimens of procedure (4) tubular occlusions were found only to a depth of 50 microm. Such occlusions were not seen in control specimens (2), in specimens where the smear-layer had not been removed (3), or following application of HEMA alone (6). It is concluded that glutaraldehyde can intrinsically block dentinal tubules. The septa in the tubules may counteract the hydrodynamic mechanism for dentinal sensitivity.