Research On The Application Of Non-diffracting Beam To Light Sheet Microscopy

The light sheet fluorescence microscopy plays an increasingly important role in real-time and fast three-dimensional imaging due to its advantages of fast speed,wide field of view,high contrast and low phototoxicity.The non-diffracting beams such as the Airy-like beam and Bessel beam have the advantages of large field of view and anti-scattering.The use of the non-diffracting beams also alleviates the mutual restriction between the field of view and axial resolution of the light sheet fluorescence microscopy.This thesis focuses on the application of the non-diffracting beams to the digital scanning light sheet fluorescence microscopy.Combined with super-resolution imaging techniques such as differential microscopy and stimulated emission depletion microscopy,the sectioning ability and imaging contrast are improved,which means that the imaging quality is further improved.The work of this thesis is shown as follows:(1)The application of sectioned Bessel beam to the fluorescent light-sheet microscopy is studied.A super-resolution technique of stimulated emission depletion microscopy combining with the light-sheet microscopy with sectioned Bessel beam is proposed to suppress the side lobes and improve the imaging contrast.In this study,the hollow Gaussian beam is used as the depletion beam.In this way,the background suppression ability of the sectioned Bessel beam in the near region of the focal plane is synergistically combined to realize the effective suppression of the side lobes in the far and near regions around the focal plane.High contrast imaging on a large field of view and wide field of view detection using low numerical aperture objective can be obtained.(2)The planar Airy-like beam based light-sheet microscopy is studied.In this thesis,the influence of parameters on the Airy-like beam is discussed.The differential imaging technology is applied to the planar Airy-like beam based light-sheet microscopy,which can effectively eliminate the background noise outside the focal plane of the Airy-like light-sheet and suppress the side-lobes of the beam.The simulation results show that the proposed method can improve the axial resolution and image contrast while maintaining a larger field of view of the Airy-like lightsheet.