| In recent years,light-sheet micro imaging technology has been studied more and more.Compared with the optical imaging methods commonly used in the experimental research,the advantages of Light-sheet micro imaging technology are obvious: large imaging depth,high contrast,fast imaging,low light bleaching and phototoxicity,high spatial-temporal resolution,etc.These are essential in life science research.Different from the traditional microscope,the illumination direction and imaging direction of the light-sheet microscope are separate and perpendicular to each other,which leads to the need for two optical paths of the light slice system: the illumination path and the detection path.The core device in the illumination direction is the galvanometer or the cylindrical mirror,which aims to form a light-sheet to excite one surface of the sample to produce fluorescence;the detection direction uses the zoom A lens or a piezoelectric translation table is used for three-dimensional imaging.Light-sheet microscope is a laser scanning microscope system,so the control system is the key part of the whole system.Its positioning accuracy and response speed have a significant impact on the measurement accuracy and speed of the light-sheet system.Due to the characteristics of Gaussian light beam,the imaging field of view will be uneven when imaging the light slice,so it is necessary to expand the uniformity of the field of view through the illumination of non diffractive light.In this paper,the light-sheet micro imaging system is discussed from two aspects of hardware synchronization control and software interface.Then,the progress of the work is shown by the results of biological experiments.Finally,the field of view is explored.The main work includes the following contents:First of all,it introduces the whole composition of the light-sheet micro imaging system and the whole design of the control system.In this paper,the x-galvanometer is used to scan the image,and then the z-galvanometer and zoom lens are used for three-dimensional imaging.Finally,the signal is collected by the SCMOS camera.The control of galvanometer and ETL is realized by pci-6713 control card,and the signal acquisition of SCMOS camera is realized by Firebird acquisition card.SLM is directly controlled by HDMI line to generate phase diagram to generate Bessel light.Secondly,the control system is introduced from three parts: the first part is the synchronous control of z-galvanometer and zoom lens to ensure accurate three-dimensional image;the second part is the synchronous control of z-galvanometer and external trigger signal of camera to ensure that the three-dimensional image with exactly one cycle is saved;the third part is the control of spatial light modulator to form a stable Bessel beam,so as to make The imaging field of view is uniform.Then,according to the imaging control system of the light-sheet system,the control software program is designed,the language is C++,vs2013 is used for development,QT is used as the framework,and the human-computer interface is designed.The image display interface and control module are integrated,and the interface is beautified.In addition,the resolution of the system is verified by the imaging experiment of the fluorescence microsphere with the built light-sheet micro imaging system,and the feasibility of the synchronous scanning control of the z-galvanometer and the zoom lens in the control system is verified.The actual measured resolution is not much different from the theoretical resolution.The imaging experiment of the zebrafish verifies the imaging speed of the system,and the zoom lens can improve the system Image speed;After the introduction of the control system and the verification of its effect,we also use the spatial light modulator to generate Bessel beam to expand the field of view.At the end of this paper,we summarize the work done in this paper,put forward the corresponding shortcomings and deficiencies,and look forward to the Light-sheet micro imaging system. |
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