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Abstract: Several fundamental problems in hydrophobic force measurements using atomic force microscope (AFM) are discussed in this paper. A novel method for colloid probe preparation based on chemical etching technology is proposed, which is specially fit for the unique demands of hydrophobic force measurements by AFM. The features of three different approaches for determining spring constants of rectangular cantilevers, including geometric dimension, Cleveland and Sader methods are compared. The influences of the sizes of the colloids on the measurements of the hydrophobic force curves are investigated. Our experimental results showed that by selecting colloid probe with proper spring constant and tip size, the hydrophobic force and the complete hydrophobic interaction force curve can be measured by using AFM.

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