Deep-tissue focal fluorescence imaging with digitally time-reversed ultrasound-encoded light

Fluorescence imaging is one of the most important research tools in biomedical sciences. However, scattering of light severely impedes imaging of thick biological samples beyond the ballistic regime. Here we directly show focusing and high-resolution fluorescence imaging deep inside biological tissues by digitally time-reversing ultrasound-tagged light with high optical gain (similar to 5x10(5)). We confirm the presence of a time-reversed optical focus along with a diffuse background-a corollary of partial phase conjugation-and develop an approach for dynamic background cancellation. To illustrate the potential of our method, we image complex fluorescent objects and tumour microtissues at an unprecedented depth of 2.5 mm in biological tissues at a lateral resolution of 36 mu mx52 mu m and an axial resolution of 657 mu m. Our results set the stage for a range of deep-tissue imaging applications in biomedical research and medical diagnostics.