By using a pH microelectrode and ultramicro-vernier shift equipment, a new method to determine directly the thickness of the sea surface microlayer (SML) at original location, is advanced and founded, which is an important innovation on the SML sampling method and the SML research. Applying the intersecting straight line method, the SML thickness was found to be approximately 60 μm. This result is consistent not only with the measurement result, (50±10) μm, obtained with the conventional glass plate method, but also with the sudden change layer of physical and chemical properties of seawater, and with the results reported in the literature. The determination method will have the ocean biogeochemistry significance on marine chemistry.