Portable microfluidic device for automatic immunofluorescence labeling and using method thereof
A microfluidic device and fluorescent labeling technology, applied in measuring devices, chemical instruments and methods, and laboratory containers, can solve the problems of difficult automation and portability, low precision, time-consuming and labor-intensive, etc., and achieve easy further Integration, improved practicability, and fast results throughout the process
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Embodiment 1
[0029] The present invention will be further described below in conjunction with the accompanying drawings. figure 1 shows the overall structural block diagram of the present invention, figure 2 Schematic diagram of the structure of the immune magnetic bead three-dimensional mixing module; it can be seen from the figure that the present invention is composed of the immune magnetic bead three-dimensional mixing module 1 and the immune magnetic bead automatic control module, wherein the immune magnetic bead automatic control module is determined by the position of the microfluidic chip The control assembly 2 and the permanent magnet position control assembly 3 are composed. In addition, there is a microfluidic chip 4 during use of the device.
[0030] Wherein, the immune magnetic bead three-dimensional mixing module of the present invention consists of an upper disc 11, a lower disc 13, an upper disc magnet 12, a lower disc magnet 14, a rotating shaft 15, a drive motor 16, a r...
Embodiment 2
[0034] The microfluidic chip described in this example is prepared by a conventional method: it consists of three plastic (such as PMMA) substrates, a bottom plate layer, a channel layer, and a sample hole layer, and an engraving machine is used to cut plastic sheets of different thicknesses and double-sided Glue, assemble in order, and heat and pressurize the assembled chip (such as 65° C., 30 min) to obtain a usable microfluidic chip.
[0035] image 3 It is a schematic diagram of the structure of the microfluidic chip, where A is the assembly diagram; B is the disassembly diagram; it consists of upper, middle and lower three-layer structure, and the middle layer contains 1 mixing pool and 5 reagent pools. Magnetic beads and various reagents required for immunofluorescent labeling (such as washing buffer, fluorescent labeling reagents, etc.); between the reagent pools are spacer pools, in which spacers are added to separate different reagents in adjacent pools.
[0036] Dep...
Embodiment 3
[0045] The device of the present invention is used to carry out automatic immunofluorescent labeling of human CD4 lymphocytes receiving different doses of ultraviolet radiation, and the irradiated cells and antibody-coated magnetic beads are added to the sample pool at the left end of the microfluidic chip; Cell fixation solution, PBS buffer solution, permeation solution (containing fluorescently labeled antibody), washing solution and immunomagnetic bead dissociation solution were added to the six liquid pools respectively; finally, spacer reagents were added to each spacer pool. According to the established procedures, the efficient mixing of immunomagnetic beads and cells and the automatic immunofluorescence labeling of cells can be completed, and the whole process is carried out at room temperature without professional operation. Figure 7 It is the radiation dose-effect relationship of CD4 lymphocytes after treatment, which shows a good linear relationship in the experimen...
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Abstract
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