A 3-DOF force sensing instrument was developed for retinal microsurgery using fiber Bragg grating sensors. Four FBG sensors were configured to maximize decoupling of axial and transverse force sensing. A nitinol flexure provided high axial force sensitivity. Experimental results demonstrated sub-millinewton resolution for axial forces and quarter millinewton resolution for transverse forces through a linear model for transverse forces and second-order polynomial model for axial forces. Validation showed the instrument can provide consistent and accurate 3D force measurement.