Device and method for separating circulating tumor cells and microembolus of circulating tumor cells by double-frequency standing wave sound field

Abstract

The invention closes a device and method for separating circulating tumor cells and microembolus of the circulating tumor cells by a double-frequency standing wave sound field. A core part of the device is composed of a flow pass base, first piezoelectric ceramic and second piezoelectric ceramic, wherein two flow passes are arranged on the upper surface of the flow pass base, and the first piezoelectric ceramic and the second piezoelectric ceramic are in contact with the bottom surface of the flow pass base, are separately located below the two flow passes, and are used for alternately generating the double-frequency standing wave sound field; the liquid outlet end of a first flow pass extends to bifurcations on both sides from a three-point-line position to form a first liquid outlet anda second liquid outlet with unequal widths, the liquid outlet end of the second flow pass extends to the bifurcations on both sides from the three-point-line position to form a third liquid outlet anda forth liquid outlet with unequal widths, and the first liquid outlet communicates with a forth liquid inlet in a smooth transition mode; and due to the fact that a cascaded two-stage channel is adopted, the alternating action of the double-frequency standing wave sound field is combined, degree of precision and accuracy of sound field separation are improved, activity of cells is effectively guaranteed, and the separated cells have large flux, unobstructed flow, high sensitivity, high integratability, good separating effect and high separating efficiency.

Description

technical field [0001] The present invention relates to the field of a separation device and separation method for circulating tumor cells and their microemboli, and in particular to a device and method for separating circulating tumor cells and their microemboli using a dual-frequency standing wave sound field. Background technique [0002] Circulating tumor cells refer to tumor cells that diffuse from tumor lesions into the peripheral blood circulation, and can develop into metastatic tumor lesions under certain conditions; while circulating tumor cell microemboli (clusters) are formed by the aggregation of two or more circulating tumor cells. As a result, more than 50% of tumor metastases are caused by circulating tumor cell microemboli, and this proportion is as high as 97% in breast cancer; thus, a rapid and effective method for separating circulating tumor cells and their microemboli has been developed. Clinically meaningful monitoring equipment provides an important b...

AI Technical Summary

Problems solved by technology

[0004] 1) Passive screening scheme: mainly based on cell size, density, surface markers and other attributes to separate, common passive screening methods include filtration, density gradient centrifugation, circulating tumor cell marker screening and microfluidic structure-based However, the filtration method and the microfluidic structure-based screening method usually cause blockage and cell damage, which affects the subsequent culture, phenotypic identification and toxicological analysis of the isolated circulating tumor cells and microemboli. The separation efficiency of density gradient centrifugation is relatively low; while the screening method of circulating tumor cell markers depends on the phenotype of circulating tumor cells, usually based on EpCAM (epithelial cell antigen) to capture and separate, but circulating tumors of different tumor sources There are differences in the expression of EpCAM in cells, and the expression of this protein will change dynamically, so there may be false positives

[0005] 2) Active screening scheme: mainly based on the effect of external force (such as electric field force, acoustic field force, etc.) to achieve separation, common methods include dielectrophoresis method and acoustic wave method; the advantage of dielectrophoresis method is that it can separate cells below 1 μm , but due to the limitation of the dielectric electric field strength, the available dielectric force is relatively small, and the separation flux is not as good as the acoustic wave method; the acoustic wave method generates a standing wave sound field in the microchannel to act on the cells to be separated. The method is more suitable for separating particles and cells with a diameter of several microns to tens of microns; according to the way of generating sound waves and the way of sound wave propagation, the sound wave method is subdivided into the surface acoustic wave method and the bulk acoustic wave method: the surface acoustic wave method is convenient for designing the sound field , but the relative sound field strength is small; the sound field strength of the bulk acoustic wave method is large, but the common bulk acoustic wave method has only a single acoustic node, which limits the separation ability of the bulk acoustic wave method

[0006] Regardless of the passive screening scheme or the active screening scheme, the separation process of the existing separation devices and methods has not been realized in the original environment where the circulating tumor cells are located, and it is difficult to effectively ensure the activity of the cells, and provide a basis for the subsequent detection of cell metabolic activity, Multiple protein marker detection, culture and toxicology research provide guarantee

[0007] Therefore, the existing technology still needs to be improved and developed

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