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This work has received funding from the European Union’s Horizon 2020 Programme through the project Advanced BiomEdical OPTICAL Imaging and Data Analysis (BEOPTICAL) under grant agreement no. 675512 and the UK Engineering and Physical Sciences Research Council (grants EP/P030017/1 and EP/R004854/1). We present the first demonstration of three-photon excitation light-sheet fluorescence microscopy. Light-sheet fluorescence microscopy in single- and two-photon modes has emerged as a powerful wide-field, low photo-damage technique for fast volumetric imaging of biological samples. We extend this imaging modality to the three-photon regime enhancing its penetration depth. Our present study uses a standard conventional femtosecond pulsed laser at 1000 nm wavelength for the imaging of 450 μm diameter cellular spheroids. In addition, we show, experimentally and through numerical simulations, the potential advantages in three-photon light-sheet microscopy of using propagation-invariant Bessel beams in preference to Gaussian beams. Postprint

Compressive sensing can overcome the Nyquist criterion and record images with a fraction of the usual number of measurements required. However, conventional measurement bases are susceptible to diffraction and scattering, prevalent in high-resolution microscopy. Here, we explore the random Morlet basis as an optimal set for compressive multiphoton imaging, based on its ability to minimise the space-frequency uncertainty. We implement this approach for the newly developed method of wide-field multiphoton microscopy with single-pixel detection (TRAFIX), which allows imaging through turbid media without correction. The Morlet basis is well-suited to TRAFIX at depth, and promises a route for rapid acquisition with low photodamage.