Axial Resolution Enhacement of Light-Sheet Microscopy via two Light-Sheets

Light-sheet fluorescence microscopy has many advantages including high-speed, noninvasive and low photobleaching and photodamage. The light-sheet thickness of light-sheet microscopy is used to determine the axial resolution. However, the light-sheet thickness is limited by the light diffraction. In order to beyond this limit, inhere, we introduce a novel way based on the use of two light-sheets to achieve the enhancement of the axial resolution of light-sheet microscopy. Two images are captured by using both Gaussian light-sheet and negative light-sheet beams. From these two images, a new relationship between them is built to achieve the axial resolution image higher than the image of Gaussian light-sheet. Experimental result is performed, indicating that the effectiveness of the proposed method is better than traditional light-sheet microscopy.