Research On Bayesian Reconstruction Microscopy Based On Super-resolution Optical Fluctuation Imaging

Currently,for the detection of active biological samples,wide field fluorescence super-resolution imaging technology is usually used.However,the traditional wide-field fluorescence super-resolution technique is limited by the imaging diffraction limit of the optical system,and usually only reaches one-half of the wavelength of the detection light.In addition,the existing wide-field fluorescence super-resolution technology usually requires tens of thousands of original image sets,and the original image utilization rate is low.At the same time,in practical applications,the optical experimental system is subject to more disturbances,the signal-to-noise ratio of the image set is low,and the existing wide-field fluorescence super-resolution technology has poor anti-noise ability.Therefore,how to solve the problem that the existing wide-field fluorescence super-resolution technology has limited resolution,low image utilization,and poor anti-noise performance is an important problem to be solved in the field of optical measurement.Aiming at the super-resolution imaging of fluctuation samples,this paper proposes a Sparse Bayesian Reconstruction Microscopy based on Super-resolution Optical Fluctuation Imaging(SBRM-SOFI),which can effectively suppress background noise including shot noise,improve image utilization and achieve super-resolution imaging..This topic has deeply studied the existing super-resolution imaging methods of fluctuation samples,including Super-resolution Optical Fluctuation Imaging(SOFI),Stochastic Optical Reconstruction Microscopy(STORM)and Sparse Bayesian Reconstruction Microscopy(SBRM),and explored the three methods in anti-noise ability,imaging speed,anti-noise ability and imaging quality.Aspects of the characteristics and improvements.Finally,the imaging resolution of SBRM-SOFI was tested experimentally.The main research contents of this topic are as follows:(1)Study the theoretical principle of super-resolution microscopic imaging methods for three fluctuation samples,such as SOFI,STORM and SBRM.The above three method characteristics are compared in terms of picture utilization,anti-noise performance,lateral resolution improvement,and image processing speed.Simulation verification of SOFI,STORM and SBRM.The parameters affecting the imaging quality of the three methods are analyzed,including the distribution density of the fluctuation sample,the duty cycle of the fluctuation sample,the signal-to-noise ratio(SNR)of the imaging system,and the size of the system point spread function(PSF).(2)The existing super-resolution imaging method of fluctuation samples is improved,and a fluctuation image imaging method with high image utilization,good anti-noise performance and high resolution is proposed,which is a SBRM-SOFI.The theoretical analysis and simulation verification of the method are carried out.(3)Establish a microscopic image acquisition system,build a super-resolution imaging system experimental platform for transmissive fluctuation samples,and analyze and process the data.When the numerical aperture(NA)is 0.65 and the fluorescence emission wavelength is 633 nm,Balanced Super-resolution Optical Fluctuation Imaging,(B-SOFI)is adopted,which can suppress the background noise,and obtain the lateral resolution of 2.6 times of the ordinary wide-field microscopy method in the fourth-order case.STORM is used to obtain a lateral resolution of 7.4 times in the particle sparse region.Using the SBRM,a lateral resolution of 5.1 times the wide field microscopy method can be obtained in the sparse region of the particle.SBRM-SOFI can effectively reduce the image noise and improve the image utilization rate,and finally obtain the lateral optical resolution of 78 nm,which is 7.6 times of the ordinary wide field resolution.