Study About Immunofluorescence Of γ-H2AX In T Lymphocyte On A Microfluidic Chip

Microfluidic chip is a kind of miniaturized and integrated technology platform. It has the ability to integrate the entire biological or chemical process into a chip that is only a few square centimeters. And it is easy to automate control. Application of microfluidic technology for real-time biomarker monitoring is a promising technology for health assessment. γ-H2AX immunofluorescence labeling of T lymphocytes can monitor the radiation damage. It can provide an important monitoring target for the evaluation of radiation damage by real-time monitoring. In order to carry out the labeling of γ-H2AX in T lymphocytes on chip automatically, this study implemented the driving of cells by microbeads, optimized labeling reaction pool on the chip and specific labeling reaction on the chip of the γ-H2AX of CD4 cells after UVC radiation.This study established the 2.8μm magnetic beads that met the magnet driven by magnetic beads on the chip, and did not affect the linear relationship between γ-H2AX relative fluorescence intensity and irradiation dose. The reaction well on chip was optimized for 6 and the number of wells between two reaction wells was 5. It also determined the reagent volume of each reaction well. The cell and bead binding ratio was 1:10, the number of cells which the experiments needs was 2×105. The research found that on chip, the cells which adsorbed microbeads could be dragged by permanent magnet. And by using this method, the γ-H2AX in CD4 cells could be labeled specifically on chip. By using different doses (0,16,32,64 J/m2) of UVC ultraviolet irradiation, this research analyzed the relationship between the relative fluorescence intensity and UVC radiation doses and concluded that the correlation coefficient was 0.9517. So it confirmed that the technology could be used to monitor the linear relationship of radiation damage and radiation dose.This research provided the basic parameters for the development of the automatic immunofluorescence staining device and micro fluidic chip.