Optical microscopy has enabled the investigation of living systems at a resolution far beyond the grasp of our bare eyes. In particular, multiphoton microscopy has allowed researchers to observe cells and biological processes under physiological conditions at depth. Refractive index inhomogeneities in tissues, however, distort the light used by such microscopes, compromising the image signal and resolution. Such distortions ultimately limit the imaging depth – and hold back our understanding of many biological processes that take place deep within living organisms.
In the Rodríguez Lab, we use a multidisciplinary approach involving physics, nonlinear optics, and microscopy development, to push the limits of in vivo deep tissue imaging. In particular, our lab works with an imaging method called three-photon fluorescence microscopy. The higher order nonlinear process involved in three-photon excitation, in combination with the reduced scattering experienced by near‐infrared excitation wavelengths, enables imaging past the limits of two-photon microscopy – the imaging workhorse in many biological laboratories. We also work with adaptive optics – a method originally developed for astronomical telescopes – to actively shape the wavefront of the light used by our microscopes, and compensate for the aberrations introduced by tissues, making it possible to achieve subcellular resolution at depth.
Our lab is especially interested in applying these technological advances to neurobiology.