Optical clearing is a versatile approach to improve imaging quality and depth of optical microscopy by reducing scattered light. However, conventional optical clearing methods are restricted in the efficiency-first applications due to unsatisfied time consumption, irreversible tissue deformation, and fluorescence quenching. Here, we developed an ultrafast optical clearing method (FOCM) with simple protocols and common reagents to overcome these limitations. The results show that FOCM can rapidly clarify 300-μmthick brain slices within 2 min. Besides, the tissue linear expansion can be well controlled by only a 2.12% increase, meanwhile the fluorescence signals of GFP can be preserved up to 86% even after 11 d. By using FOCM, we successfully built the detailed 3D nerve cells model and showed the connection between neuron, astrocyte, and blood vessel. When applied to 3D imaging analysis, we found that the foot shock and morphine stimulation induced distinct c-fos pattern in the paraventricular nucleus of the hypothalamus (PVH). Therefore, FOCM has the potential to be a widely used sample mounting media for biological optical imaging. optical clearing | tissue clearing | deep tissue imaging

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The doughnut beam is a spatially structured beam which has been widely used in super-resolution microscopy, laser trapping and so on. However, when it passes through thick scattering medium, aberrations will seriously affect its performance. Currently, adaptive optics (AO) has become one of the most powerful tools to compensate aberrations. However, conventional AO always suffers from limited corrected field of view (FOV). Here, we propose a method with conjugate AO system based on coherent optical adaptive technique. The results show that the corrected FOV can be improved effectively. For a wide range of the optical applications with doughnut beam, our method has potentials in correcting aberrations with high speed in turbid media.(A) Mouse brain slice, (B) the distribution of r PAO , (C) the distribution of r CAO . The vortex beam focus of the blue point in (B) and (C) among a 137.5 × 137.5 μm FOV (D1) ideally, (D2) with scattering, (D3) in pupil AO system and (D4) in conjugate AO system. K E Y W O R D S aberration correction, adaptive optics, doughnut beam, turbid medium