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Micro-fluidic chip, manufacturing method thereof and fetus erythroblast capturing and releasing method

A microfluidic chip, nuclear red blood cell technology, applied in the field of molecular cell biology detection, can solve problems such as impact detection and analysis, fetal genetic disease detection and analysis obstacles, purity as low as 20%, etc., to achieve high efficiency and high purity The effect of separation, reduction of non-specific adsorption, and simple preparation process

Active Publication Date: 2018-11-13
SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although about 99.99% of red blood cells and 99.90 to 99.99% of white blood cells can be removed by a two-stage microfluidic chip, there are still 5,000,000 red blood cells and 8,000 to 8,000,000 white blood cells in the enriched sample, which will also affect downstream detection and analysis
[0009] As another example, Document 2 uses red blood cell hyperaggregation (hyperaggregation) and red blood cell lysis to remove red blood cells, and then enriches fetal nucleated red blood cells through a micro-magnet array. It can also capture dozens to hundreds of fetal nucleated red blood cells per milliliter of blood. But the purity may still be as low as 20%, creating obstacles to the detection and analysis of fetal genetic diseases

Method used

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preparation example Construction

[0058] In some embodiments, the preparation method includes: processing the second substrate to form a second microstructure part matched with the first microstructure part.

[0059] In some embodiments, the preparation method includes: providing a second mold with a second set structure, and using the second mold to form a second microstructure part corresponding to the microchannel. substrate.

[0060] More preferably, there are more than two microstructures distributed in the microchannel, and the distance between adjacent microstructures is sufficient to allow fetal nucleated red blood cells to pass through, and at least one of the microstructures also passes through the connecting arm The capture substance is immobilized.

[0061] In some embodiments, the preparation method includes: bonding the first substrate and the second substrate to form the microfluidic chip, and the bonding method includes thermal bonding, solvent-assisted bonding, and solvent-assisted bonding. ...

Embodiment 1

[0087] Reference 3-Document 5, preparation of glass-PDMS (polydimethylsiloxane) based microfluidic chip, in which the microfluidic channel array is 50 with a width of 30 μm, a depth of 150 μm, and a length of 20 mm, in a sinusoidal shape Composed of micro-channels, the micro-channel arrays are distributed in parallel between two main channels with a width of 1.5mm in a zigzag distribution. After that, plasma cleaning is used to activate the glass and PDMS surface in the microfluidic chip, and then bonded, and then (3-aminopropyl) triethoxysilane (APTES) is used to modify the surface of the microfluidic channel, and the connection can be detected by light. After cleaved biotin (photocleavable biotin (NHS-PC-Biotin)), streptavidin and biotinylated CD71 antibody were connected sequentially to capture and isolate fetal nucleated red blood cells.

[0088] The process is as follows: 100 μL of 2% (3-aminopropyl)triethoxysilane (APTES) ethanol (95%) solution was flowed through the mic...

Embodiment 2

[0090] Reference 3-Reference 5, preparation of glass-PDMS (polydimethylsiloxane) based microfluidic chip, in which the microfluidic channel array is 16 with a width of 200 μm, a depth of 100 μm, and a length of 20 mm, in a straight line The micro-channels are composed of 16 micro-channels, which are divided into 16 micro-channels from the inlet and outlet respectively, and there are many micro-columns distributed in the micro-channels. The height of the micro-columns is 100 μm, and the diameter is 20 μm. The distance is 20 μm.

[0091] After that, plasma cleaning was used to activate the glass-PDMS surface, and the microchannel surface was modified with aminosilane APTES, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N- Hydroxysuccinimide (NHS) linked to 4-{4-[1-(9-fluorenylmethoxycarbonamido)ethyl]-2-methoxy-5-nitrophenoxy}butanoic acid (Fmoc- Photo-Linker, after (4-{4-[1-(9-Fluorenylmethyloxycarbonylamino)ethyl]-2-methoxy-5-nitrophenoxy}butanoic ...

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Abstract

The invention discloses a micro-fluidic chip. The micro-fluidic chip comprises a micro-channel and a capturing substance, wherein the micro-channel at least allows target substances such as fetus erythroblasts to pass through; the capturing substance can be specifically combined to the target substances; the capturing substance is fixed in the micro-channel via a connecting arm; the connecting armcomprises at least one locus which can be cut by light; and after more than one locus which can be cut by light is cut by light with selectable wavelength, the connecting arm is broken completely. The invention further discloses a manufacturing method of the micro-fluidic chip and a fetus erythroblast capturing and releasing method. by the micro-fluidic chip, fetus erythroblasts and the like canbe captured specifically, high-efficiency and high-purity separation of the fetus erythroblasts is realized, the captured fetus erythroblasts can further be selectively released on monocyte level in amode of light cutting and the like, and follow-up analysis is facilitated. Meanwhile, the micro-fluidic chip is low in manufacturing cost and simple in process.

Description

technical field [0001] The present invention relates to a microfluidic chip, its preparation method and application, in particular to a microfluidic chip capable of specifically capturing fetal nucleated red blood cells and its preparation method, and its application in fetal nucleated red blood cell capture and single cell The application in release belongs to the technical field of molecular cell biology detection. Background technique [0002] Fetal nucleated red blood cells (fNRBC) in the peripheral blood of pregnant women refer to fetal cells that enter the maternal systemic circulation through the placental barrier. There are no nucleated red blood cells in the peripheral blood of normal adults, so after excluding blood system diseases, the nucleated red blood cells found in the peripheral blood of pregnant women should come from the fetus. Fetal nucleated erythrocytes persist in maternal blood during pregnancy, and their lifespan is generally less than 90 days, so th...

Claims

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Application Information

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IPC IPC(8): C12M1/00B01L3/00C12N5/073
CPCC12N5/0603C12N5/0641B01L3/50273B01L3/502761B01L2300/0887B01L2300/0864
Inventor 樊丽王宏
Owner SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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