3D Reconstruction Technology For Bright Field Microscopic Optical Sectioning

The traditional biological microscopic images only can display the 2D structure or morphological distribution of the biological tissue,but the true form of biological tissue is 3D structure,which is difficult for judging the relationship with structure and function of tissue.As the technique of microscopic sectioning and the related equipment invented,it provides the possibility for 3D reconstruction of the biological tissue.At present,many optical sectioning techniques all are based on fluorescence microscopy imaging,such as laser scanning confocal microscope,structured illumination microscope,plane illumination microscope,two-photon microscope and so on.It has limited the widely application as the equipment is complex,expensive and difficult to operate.Therefore,it can't replace the bright field microscope which is relatively simple,easy to operate and widely used.It has great practical value to make the research for optical sectioning technique about non fluorescence and bright field.Based on the traditional bright field microscope,the axial resolution of the microscope is increased by designing the special illumination method.We obtain the bright field optical sectioning images with the thickness about 600 nm.It is a key step for optical sectioning technique to 3D reconstruction by using a series of optical sectioning images.However,the contrast ratio of the bright field microscopic optical sectioning was low,which brings great difficulty for the 3D reconstruction.Focusing on the image features of bright field microscopic optical sectioning,the paper made the deep research for the 3D reconstruction technique.As the contrast of bright field microscopic optical sectioning is low,the optical sectioning images need to be processed by using many kinds of image processing techniques,the appropriate image processing techniques were applied through analysis and comparison.Image segmentation technique is proposed to extract the interesting part from the optical sectioning images.Combining the features of the images,the marker-controlled watershed method was proposed to achieve extracting.Through using image processing techniques about hole filling and morphology and so on,the segmentation results have gotten optimization.By analyzing a variety of 3D reconstruction algorithms,the Marching Cube algorithm is selected to achieve the 3D reconstruction of bright fieldmicroscopic optical sectioning.Finally,a graphical user interface based on the 3D reconstruction procedure of bright field microscopic optical sectioning was design with multiple functions through graphical user interface development environment provided by Matlab.The graphical user interface is good to make the technique practically.