High axial resolution and long field of view for light-sheet fluorescence microscopy via double-beam aperture

Abstract

We propose a simple method based on the use of interference of the double-beam aperture to enhance both the axial resolution and field of view of light-sheet fluorescence microscopy. The double-beam aperture placed in the pupil plane generates multiple-spot intensity patterns in which the size of central lobe reduces. By scanning this intensity pattern along x -axis, the light sheet is generated. By satisfactorily choosing the numerical apertures of illumination lens and detection lens, only the central light sheet is used to achieve image, so the axial resolution of light-sheet fluorescence microscopy is enhanced. Both the numerical apertures of the illumination lens and detection lens of 0.3 and 1.1, respectively, are employed to perform the simulation results. The simulation results indicated that both the axial resolution and field of view are improved in comparison to the Gaussian light-sheet. Additionally, in order to remove a small amount of the existing outside lobes, we propose a subtraction method. The simulation results demonstrated that our technique can eliminate the outside lobes in the system point spread function of the double-beam aperture beam light sheet.