Research On Weak Fluorescence Image Analysis And Rapid Imaging System Of Confocal Laser Scanning Microscopy

Confocal Laser Scanning Microscopy(CLSM)is a high-tech for studying submicron tissues.With the advantages of high resolution and non-destructive slicing technique,CLSM has great application value in the fields of biology,medicine and industry.However,there are many weak fluorescence images in the medical field.Traditional CLSM often exists many technical problems,including taking a long time in the process of manual focusing,the slow speed of galvanometer scanner,owning small field of view,blurred image of the system.Therefore,it is necessary to integrate the image analysis technology and the system hardware design.On the one hand,the resolution of the weak fluorescence sample image is improved,and on the other hand,the imaging speed is increased.This article focuses on the following aspects: accelerating the autofocusing of dark-field images,improving the stitching of weakfluorescent images,and restoring confocal slices with poisson noise.(1)Aiming at the dark field problem of confocal image,the fast and robust autofocus technology of weak fluorescence image is studied.Better autofocus evaluation function can be obtained by combining the gray image and color image features.(2)Considering the contradiction between high resolution and small view field of CLSM,the stitching technology of low-contrast and weak fluorescence images is studied.The method of combining image enhancement and SURF feature extraction can be used to accelerate image stitching process.(3)To solve the problem of noise and blur in three-dimensional slice of confocal microscope,a spatial adaptive high-order total variation method is applied to conduct image restoration.(4)Concerning the long time for image acquisition and lack of accuracy of the collected images,the accelerating technology of the confocal microscope system is studied.Fuzzy PID control can be used to improve the motion accuracy of the image acquisition platform,and the nano Z-axis is realized by using piezoelectric ceramics.By the combination of resonant galvanometer and galvanometer galvanometer,the scanning process takes less time.This study has certain significance for rapid image acquisition and precise imaging of confocal microscopes.