Cell Micro Scanning Analysis System Based On Photonic Crystal

As an unlabeled "biomarker" for real-time expression of cell physiological state,cellular mechanical information can effectively reflect the stress pattern of interaction with external matrix during cell culture.Cellular mechanical information is a powerful analysis basis for cell homeostasis,migration,proliferation,and apoptosis.Currently,traction microscope and microcolumn array are commonly used to measure cell force,which has the inherent disadvantages of a large amount of inverse calculation,few observed cells and the introduction of unnatural substrate mode.Photonic crystal material films have structural colors and good biocompatibility.Different colors can be produced by changing lattice spacing and refractive index.This design utilizes the color change characteristics of cells under mechanical stress on photonic crystal film to build a new real-time unlabeled cell micro scanning analysis system based on a photonic crystal.The specific research work is as follows:1)Build an infinite conjugate microscope for photonic crystal imaging through optical design.Aiming at the sensitivity of the Angle of incident light in a photonic crystal,and the problems of color mixing and color rendering caused by the improvement of the Angle non-parallelism caused by common coaxial illumination passing through the high-power objective lens,collimating light source,and coupling device were designed to improve the parallelism of the output light of the objective lens.It provides an optical scheme for observing cell stress by using photonic crystal materials.2)In view of the complexity and precision of manual displacement and focusing operation in microscopic observation,an embedded control three-axis displacement platform is built by using NVIDIA Jetson Nano.The visual field movement and focusing can be accomplished by electronically controlled mechanical slide,and the focusing accuracy can reach 0.07 mm.It has the function of large-field displacement observation and long-time timed autofocus acquisition,and can simultaneously collect the bright field image of cell imaging by transmitted light and the photonic crystal image by reflected light.Through the circuit and mechanical structure design,the miniaturization and integration of the equipment can be placed in the incubator for real-time dynamic observation.3)In view of the application scene requirements of microscopic images that are limited by the field of vision and are difficult to directly obtain large-field images,a scanning focusing stitching algorithm is studied to quickly obtain high-resolution large-field images.The focal plane is modeled through 9-point scanning and the focus is dynamically pursued during the scanning process.The whole 96-well plate can be collected within 6 minutes to complete scanning and splicing,and clear patterns with 4K resolution in the hole plate can be obtained.4)Finally the device is proposed to calculate the cell analysis application deployment,mainly including cost function method based on modeling the cell points matching of position tracking,fluorescent cells microspheres foreground segmentation feature modeling method and YOLOv5 s lightweight model training method of Ming field of epithelial cells of complicated background motion detection work.