Axial resolution enhancement for light sheet fluorescence microscopy via using the subtraction method

Light sheet fluorescence microscopy is an imaging method in which the sample is illuminated from the side by a wide and relatively thin light sheet. This method minimizes the out-of-focus excitation, provides a good balance between spatial resolution and temporal resolution, and mitigates photobleaching and photodamage. Unfortunately, the axial resolution of light sheet fluorescence microscopy is much lower than lateral resolution, which limits the achievement of isotropic imaging. We propose to combine the subtraction method with light sheet fluorescence microscopy to improve the axial resolution. The experimental results demonstrate that the proposed method can produce higher axial resolution than that of conventional light sheet fluorescence microscopy. In addition, a way to reduce negative values in subtraction method is also discussed.