2 results about "Total internal reflection fluorescence microscope" patented technology

The application discloses a new method for quantitatively analyzing interface nucleic acid hybridization reaction and application thereof, and comprises the following steps: S1, constructing a DNA tetrahedron interface; S2, imaging the DNA tetrahedron interface through a total internal reflection fluorescence microscope; S3, realizing real-time monitoring of a hybridization event of the DNA tetrahedron interface by adopting double-color co-localization and analyzing a fluorescence signal; and S4, analyzing hybridization kinetics on the DNA tetrahedron interface by recording changes of the fluorescence signal with time. The application uses the DNA tetrahedron interface for single-molecule fluorescence imaging, can realize real-time monitoring of a single nucleic acid molecule hybridization event, improve detection sensitivity and specificity, accurately analyze kinetic parameters of a hybridization reaction, help realize more accurate performance evaluation of a nucleic acid sensor, has important significance for design and optimization of the nucleic acid sensor, and thus plays a greater role in the fields of clinical diagnosis, environmental monitoring and food safety.

This invention discloses a method for the separation and detection of exosomes in whole blood using a microfluidic chip. The method includes preparing a microfluidic chip with a specific structure for whole blood exosome separation and a PEGylated microfluidic chip with fishbone-like protrusions for capturing whole blood exosomes. Simultaneously, a viscoelastic solution is prepared and the whole blood sample is pretreated. The pretreated whole blood sample is injected into the microfluidic chip for whole blood exosome separation, and size-dependent separation of exosomes is achieved using a viscoelastic flow field. After concentration, a whole blood exosome concentrate is obtained. Exosomes are immobilized with paraformaldehyde, perforated, and biotinylated. After capture, a BODIPY polymer dot-coupled secondary antibody is prepared, combined with a primary antibody to complete exosome immunolabeling. After localization, multi-channel imaging using a total internal reflection fluorescence microscope is performed. This method achieves efficient separation and highly specific labeling detection of whole blood exosomes, improving the separation recovery rate and purity.