Design And Development Of High-Content Cell Culture Software System

The successful development of new anti-tumor drugs can bring great social and economic benefits.Traditional methods for the research of anti-tumor drugs based on animal model have the disadvantages of high cost,long testing period and low effectiveness.As an emerging in-vitro culture technique,tumor sphere can not only save cost and reduce testing cycle,but also realistically simulate the adhesion,invasion and distant metastases of tumor cells.Therefore,it is widely used in research and development of anti-tumor drugs.In the development of anti-tumor drugs using the spheroid model,high-content cell culture systems are needed to achieve real-time observation and analysis of cell spheres,thus providing reliable data support for anti-tumor drug development.However,in view of the specific scene of the tumor cell sphere,existing high-content cell culture systems have the problem of functional waste.Besides,the price of the system is very expensive.In order to better carry out the pharmacodynamic analysis of tumor drugs,it is urgent to develop a high-content cell culture system with low cost and satisfying core needs.This paper aims to design and develop a high-content cell culture software system to facilitate researchers to conduct a series of operations,such as the culture,observation,image acquisition and automatic calculation of morphological parameters of tumor cells.The hardware of the whole high-content system includes the cell culture incubator with the function of environmental control,inverted fluorescence microscope,CCD camera for image acquisition and PC end.The software is mainly divided into five modules,in which the incubator module is used to configure and monitor various environmental parameters(including various gas concentrations,temperature and humidity,etc.)during the culture process;the optical platform module is used for automatically positioning the cell;the camera module is used for real-time display and storage of cell images or videos;morphology parameter calculation module is used for autonomic calculation of common morphological parameters of cells;database storage module is used to store configuration,motion and morphological parameters utilized in the above four modules.In order to reduce the coupling among modules and increase the maintainability,the system architecture is divided into display layer,business logic layer,interface layer and underlying service,in which the display layer provides a friendly user interface for high-content systems;the business logic layer implements the standardized management of tasks;the interface layer encapsulates the basic tasks of each module and integrates the camera flow;the underlying service realizes the communication between the upper and lower computers through the custom serial protocol and communication function.The high-content cell culture software system developed in this paper also has the function of automatically calculating the morphological parameters of the cell sphere,which plays an important role in the quantitative analysis of the cell sphere.The main steps in the calculation of morphological parameters are: establishment of sample library,cell sphere segmentation,calculation of morphological parameters,in which the cell sphere segmentation is the core of morphological parameter calculation.In this paper,four traditional methods(including adaptive threshold,OTSU,HSV and Kmeans)are used to segment the cell sphere image.On the basis of accurate segmentation,the automatic calculation of the morphological parameters(including cell radius,area,roundness and mobility)is implemented.Considering that the system will face various tumor scenes in the course of clinical use,and more types of tumor images will be obtained.In order to improve the scalability of the segmentation algorithm and match the future needs of the high-content cell culture system,the U-Net deep learning image segmentation method is further explored in this investigation.Finally,the high-content cell culture system is tested in terms of function and performance to ensure the stability in practical applications.