As biomedical research delves deeper into the intricacies of tissue structure and function, the demand for high-resolution, high-signal-to-noise-ratio deep-tissue imaging technologies has become critical. Traditional microscopy, limited to two-dimensional, transparent thin biological samples, falls short in satisfying the current requirements for three-dimensional deep-tissue imaging in biomedical sciences. In contrast, light-sheet fluorescence microscopy distinguishes itself through its attributes—low photodamage, rapid acquisition, extensive field of view, and volumetric imaging—establishing it as a cornerstone among biologists. Despite these advantages, the intrinsic high-scattering characteristics of biological tissues pose formidable challenges to achieving deep imaging. This review focuses on recent progress in light-sheet fluorescence microscopy for deep tissue imaging, emphasizing strategies that overcome challenges associated with high-scattering samples, so as to provide valuable insights to researchers in related fields, assisting them in developing a preliminary comprehension of the latest innovations and future possibilities inherent in this cutting-edge technology. Firstly, the basic principles of light-sheet fluorescence microscopy and the causes and effects of high-scattering absorption characteristics are explained. Subsequently, recent progress in enhancing tissue penetration depth and addressing issues such as light scattering and absorption is further examined. Finally, the development prospects and potential applications of light-sheet fluorescence microscopy imaging technology with high penetration depth and anti-scattering ability are discussed.