A technique for accurate measurement of the principle refractive indices and birefringence for silklike samples is presented. It is based on rotating the linear polarization of the illuminating light on a silk immersed in reference liquid to achieve index matching at the silk/liquid interface. The technique was used to measure the principal refractive indices of a P. eburnus radial silk at different strains. This in turn allows the calculation of strain-optic coefficients. The first measurement of the strain-optic coefficients of spider silk is presented. The technique is more generally applicable to strain-optic study of birefringent micro-optic samples.