Research On Human Lymphocyte Capture And Radiation Damage Assessment Technology Based On Gold Coated Magnetic Beads

With the rapid development of today's science and technology,many countries have nuclear weapons and nuclear energy.While safeguarding national security and providing economic growth,they also bring certain nuclear threats.In recent years,in the case of frequent occurrence of various nuclear accidents,the rapid estimation of the radiation dose received by humans is an important reference for current radiation therapy and diagnosis.In this paper,the kinetic analysis of lymphocytes captured by immunomagnetic beads,human lymphocyte capture based on gold-shell magnetic beads and the quantitative analysis of the immunofluorescence intensity of?-H2AX based on flow cytometry are studied.The main research contents are as follows:1.An analytical model of the magnetic magnetic beads and human lymphocyte complexes in the magnetic field and flow field was established.The formulas for the magnetic field force and the flow field force were derived.The Y-axis magnetic field force component of a single magnetic bead was 0-50pN.The flow field force experienced by the composite in the flow field is 0-527.5 pN.Secondly,the simplified simulation model is established by using COMSOL software.The simulation of the motion trajectory is completed by the steps of physical field coupling,mesh division and solver setting.Finally,the effects of four parameters of permanent magnet position,inlet initial flow velocity,bead radius and permanent magnet remanence on the capture effect of the composite were analyzed.It was found that when the initial flow velocity of the inlet was 3 mm/s,the radius of the magnetic bead was 500 nm,and the permanent magnet remained.Magnetic resonance of 0.8T has a good capture effect on the complex of immunomagnetic beads and human lymphocytes,laying a theoretical foundation for cell capture experiments.2.The sol-gel method was used to complete the controllable preparation of magnetic microspheres(500 nm),and the gold-shell magnetic beads coated with gold shells were successfully prepared by the ultrasonic-hydroxyamine seed growth method proposed by our group.At the same time,the CD3 antibody was coupled to the gold-shell magnetic beads using a self-assembled monolayer membrane technique.The SU-8 gel film was prepared by spin coating,photolithography and development.The preparation of two microfluidic chips was completed by casting,curing and bonding.Human peripheral blood lymphocytes were counted and stained,and a capture experiment was conducted.The number of lymphocytes was counted by the cell counting plate before the chip was introduced and the waste liquid was collected,and the capture efficiency was about 60%.And the stained lymphocytes can stimulate the fluorescence energy under the fluorescence microscope to visually observe the capture of cells in the chip.3.The antibody pre-experiment was completed.After the?-H2AX antibody specifically binds to the?-H2AX antigen during antibody labeling,the goat anti-mouse FITC fluorescent antibody specifically binds to the mouse?-H2AX antibody,so that?-H2AX is A fluorescent signal is generated.Immunofluorescence signal analysis was performed on lymphocytes and lymphocytes captured by magnetic beads by flow cytometry.The linear regression equations for successfully obtaining different radiation doses and their respective average fluorescence intensities are Y=4298.24x+20036.62,R~2=0.9996,Y=3142.93x+2073.2.24,R~2=0.7584.