Towards Real-time Detection Of Glucose Consumption Of Embryos Using A Microfluidic System

Assisted Reproductive Technology(ART)provides the possibility of treatment for infertility,but it still faces problems such as low birth rate and poor development after birthing.Screening a single embryo with high quality and high developmental potential for transplantation is the key to solving this problem.The current screening method is the morphological analysis established by Prof.Gardner,which relies on the subjectivity of doctors and is difficult to standardize.Embryo metabolism is directly related to the quality of the embryo and the developmental potential of the embryo,and can serve as a new standard for embryo screening.The microfluidic chip enables high-precision quantitative detection of embryo metabolites,and establishes a stable embryo culture environment and detection environment in the channel.In this paper,we focus on the detection of glucose metabolism,and replace the embryo with human umbilical vein endothelial cells(HUVEC)to establish a microfluidic system for cell culture and glucose detection.This thesis designed a microfluidic chip for cell culture and glucose fluorescence detection.A microfluidic chip was used as the culture and reaction device,and the whole electric inverted fluorescence microscope and photomultiplier tube were used as the detection device to detect the glucose consumption of the cells during the culture process.The dilution of the medium in the chip and its mixing with the indicator were studied by numerical simulation.It was verified that under the design conditions,when the solution in the chip flows through the detection zone,it has been uniformly mixed to avoid the influence of uneven concentration distribution on the detection.By numerical calculation,the change of glucose concentration when the cells were cultured in two ways,the continuous injection medium and the flow-stop method were provided.The concentration of glucose changed little when cultured cells were continuously injected,which was difficult to detect.In contrast,cells were cultured in a "Flow-Stop" protocol with conditions of injecting 20 ?L medium at a flow rate of 1 ?L·min-1 every 2 h.In this thesis,long-term culture and glucose detection of cells on microfluidic chips were achieved.A glucose fluorescence detection system with Amplex Red as an indicator was established.The dynamic response range was 20?300 ?M,the minimum detection limit was 10 ?M,and the glucose consumption of cells was obtained within 24 hours.The system can realize glucose detection of cells,and provides an effective method for multiple detection of embryo metabolites in the future.