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Non-invasive optical focusing inside scattering media is still a bigchallenge because inhomogeneous media scatter both incoming photons forfocusing and outgoing photons for observation. Various approaches, utilizingnon-linear fluorescence or ultrasound, have been reported to address thisdifficulty. However, implementation of these methods is complicated and highlyexpensive, as ultrafast laser systems or photo-acoustic equipment must beemployed. Here, we demonstrate a wavefront shaping technique to achievenon-invasive focusing (NiF) inside scattering media using only a linearfluorescent signal. Contrast and mean of incoherent speckles, produced by thelinear fluorescence, are utilized as feedback signals to optimize the inputwavefront. While increasing speckle contrast makes the focus tighter, andincreasing the speckle mean enhances the intensity, fine-tuning thecontribution of these two factors in our two-step optimization is essential. Anoptimal wavefront is found to achieve simultaneously both a micrometer focalspot size (down to 20 um diameter) and high intensity (more than a 100-foldenhancement) inside the scattering media. Our method promises a new route inlife science towards focusing, imaging or manipulating deep into biologicaltissues with linear fluorescent agents.

Non-invasive optical focusing inside strongly scattering media with linear fluorescence