Separate detection system and method of rare cells based on centrifugal micro-fluidic technology

Abstract

The invention discloses a separate detection system and method of rare cells based on a centrifugal micro-fluidic technology. The system comprises a micro-fluidic chip similar to an optical disk, a centrifugal drive module and an optical detection module, wherein the micro-fluidic chip comprises a plurality of groups of micro-channels and micro-cavities arranged in a radiation manner; and the entire chip structure is composed of an elastic micro-column guide rail layer, a deformable film layer, a pipeline / cavity layer, a filtering membrane layer and a liquid waste collecting layer. The method comprises the steps of: firstly, leading a sample solution and immune modified microspheres into a liquid storage cavity through an injection port of the micro-fluidic chip in use, putting the sample solution and the immune modified microspheres on a centrifugal platform of the centrifugal drive module, assembling an elastic micro-column, and rotating at a low speed, so as to achieve fully mixing and reacting of the sample solution and immune modified microsphere liquid in the liquid storage cavity, separating by a high-speed rotary chip, and then dropwise adding a specifically recognized fluorescently-labeled antibody solution in each separate cell collection area, carrying out incubate reaction, adding a buffer solution and centrifuging, and finally identifying and analyzing through the optical detection module.

Description

technical field [0001] The invention relates to a rare cell separation and detection system based on centrifugal microfluidic technology and its application method, which can be applied to the fields of non-invasive prenatal diagnosis, early tumor diagnosis and prognosis monitoring, stem cell research and the like. Background technique [0002] Selective isolation of rare cells from complex mixtures is of great significance in both cell biology and clinical aspects, such as isolation of fetal red blood cells from maternal peripheral blood, isolation of circulating tumor cells (Circulating Tumor Cell, CTC) from peripheral blood of patients, and bone marrow or Separation of stem cells in tissues, etc. The analysis of these cells can often provide very valuable information for clinical diagnosis and treatment of diseases, but the content of such cells in samples is very rare, usually only 1-10 cells / mL, separation and extraction very difficult, thus greatly limiting its clinica...

AI Technical Summary

Problems solved by technology

Nevertheless, this method still has major deficiencies: on the one hand, in order to ensure the capture efficiency of cells, the blood sample must maintain a relatively slow speed (~1mL / h), so the detection process of this method takes a long time (5~7 hours), and the efficiency is low

On the other hand, the chip used is made of opaque silicon base, which is not conducive to optical detection, and the plasma etching process of the micropillars in the chip requires expensive instruments and high production costs, which limits its large-scale clinical application

Although, some people have tried to improve this method recently [Wang S, Liu K, Liu J, et al.Highly Efficient Capture of Circulating Tumor Cells by Using Nanostructured Silicon Substrates with Integrated Chaotic Micromixers.Angew.Chem.Int.Ed.2011, 50:3084–3088.], such as optimizing the capture efficiency of CTCs in microfluidic chips by adjusting the spacing and arrangement of immune capture micropillars or adding oblique ridge microstructures in microchannels, but the improvement effect is limited, and it still cannot meet the requirements of efficient capture at the same time. and quick separation requirements

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