In this paper, we present our experimental results on the process of the in situ chemical modification of the silicon nitride atomic force microscopy/frictional force microscopy tip by n-octadecyltrimethoxysilane (OTE)/mica, OTE/SiO2, and SiO2. The modified tips have different friction and adhesion properties against mica reference samples compared to those before modification. The resultant tip modification depends not only on OTE self-assembled monolayer (SAM) but also on the substrates the OTE SAM is prepared on. In the case of OTE/mica, the friction of the modified tip against mica reference is greatly reduced; in the case of OTE/SiO2, the friction of the modified tip against mica reference is greatly increased. It is surprising that bare SiO2 can also chemically modify the Si3N4 tip to increase the friction against mica reference. In the case of OTE modification, it was found that the tips could be cleaned by repetitive friction scans on mica. However, a tip modified by SiO2 cannot be mechanically cleaned. Moreover, it was found that humidity and load could also affect the tip chemical modification. Ourresults are important for interpreting tribological data since the actual contact chemistry was often overlooked in the atomic force microscopy experiments in the past.