![]() The intensity of single-scattered waves used in conventional imaging decreases exponentially with depth thus, the imaging depth limits are set by the detector dynamic range and the efficiency of the gating operations.Īpproaches that make deterministic use of the abundant multiple-scattered (MS) waves have been proposed to enable deep optical imaging while maintaining the microscopic spatial resolving power. ![]() Optical microscopy is an indispensable tool in biology and medicine owing to its high spatial resolution, molecular specificity and minimal invasiveness, but it is limited to the interrogation of superficial layers for in vivo imaging. In this Review, we provide an overview of recent developments in this area, with a focus on approaches that achieve a microscopic spatial resolution while remaining useful for in vivo imaging, and discuss their present limitations and future prospects. New approaches have emerged based on the deterministic measurement and/or control of multiple-scattered waves, rather than their stochastic and statistical treatment. To achieve an imaging depth beyond this conventional limit, there is increasing interest in exploiting the physics governing multiple light scattering. Adaptive optical microscopy, which minimizes the effect of sample-induced aberrations, has to date been the most effective approach to addressing this problem, but its performance has plateaued because it can suppress only lower-order perturbations. This problem originates primarily from multiple light scattering caused by the inhomogeneity of tissue obscuring the desired image information. ![]() However, its working depth for in vivo imaging is extremely shallow, and thus reactions occurring deep inside living specimens remain out of reach. Optical imaging has had a central role in elucidating the underlying biological and physiological mechanisms in living specimens owing to its high spatial resolution, molecular specificity and minimal invasiveness.
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