Binding Quantum Dot To GLUT4 In Live Cells

Glucose Transporter4(GLUT4) palys an very important role during the transportation of glucose in live cells. Understanding the dynamics of GLUT4 not only that it is very useful to the treatment of some diseases(e.g. diabetes), but also that it help us to know some basic life phenomenon clearly.As a new type of fluorescent probe, quantum dots are widely used in the life sciences recently. Because of its broad absorption spectrum, narrow emission spectra, emission stronger and higher stability and other unique advantages, quantum dots are rapidly used in the field of life science research.The L6 cells were used as the investigate material. Combined with wide-field fluorescent microscopy, total internal reflection fluorescence microscopy, confocal fluorescence microscopy and other platforms, the quantum dots were bound to GLUT4 specially by immunological technique. At first, judged the feasibility of immunological technique which was used to bind with the GLUT4 molecules of fixed cells; Then, discussed the advantages and disadvantages of the different binding methods between the living cells and quantum dots, just because it is useful to find out the best way which is fit for the condition, and it can help to research some life phenomenon. Finally, chose the method of binding IgG-Qdots to GLUT4 molecules of living cells in 4℃, combined with green fluorescent protein technology and MATLAB analysis tools, research the transport of GLUT4 vesicles from the whole-cell level and the single-particle level respectively,. Compared the experimental data with other researchers'conclusions, the quantum dots seems have no harmful effect on the normal physiological activities of GLUT4 vesicles. This indicates that the binding method can be applied to study the detailed transport mechanism of GLUT4 vesicles. In addition, the method of bind quantum dots with biomolecules can be used in some other biological studies.