By adapting femtosecond micromachining approaches developed in hydrogels, we can perform Intra-tissue Refractive Index Shaping (IRIS) in biological tissues. We reduced femtosecond laser pulse energies below the optical breakdown thresholds to create grating patterns that are associated with a change in the refractive index of the tissue. To increase two-photon absorption, we used a two (or more)-photon-absorbing chromophore.