A method for preparing a microfluidic chip embedded with zwitterionic functionalized nanofiber membranes

A nanofiber membrane and microfluidic chip technology, which is applied in the field of microfluidic chips and cell sorting, achieves the effects of simple processing technology, rapid non-destructive release, and easy detection and analysis.

Active Publication Date: 2020-11-13
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although this principle has not been applied to the release of larger particles, cells, especially CTCs, based on previous research, we can use weakly acidic phosphate buffer to treat targeting ligands containing phenylimide groups in vitro. body to achieve rapid and non-destructive release of CTC

Method used

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  • A method for preparing a microfluidic chip embedded with zwitterionic functionalized nanofiber membranes
  • A method for preparing a microfluidic chip embedded with zwitterionic functionalized nanofiber membranes
  • A method for preparing a microfluidic chip embedded with zwitterionic functionalized nanofiber membranes

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] (1) Add 0.5g of polyethyleneimine PEI and 1.5g of polyvinyl alcohol PVA into 18g of deionized water, stir magnetically at 90°C for 4h to obtain a PEI / PVA spinning solution with a mass fraction of 10wt%, and draw it into a 10mL syringe For PEI / PVA spinning solution, the syringe is connected to a 16-gauge needle, and the flow rate of the syringe pump is set to 0.6mL / h. The receiving device is placed on a round cover glass or a rectangular glass slide placed flat on the aluminum foil paper, and the receiving distance is adjusted to 20cm. , set the spinning voltage to 28.5kV, and carry out electrospinning treatment under the conditions of ambient temperature 25°C and humidity 40%, to obtain PEI / PVA nanofiber membrane, put it into a vacuum dryer for steam cross-linking, and place it at the bottom of the vacuum dryer There is a glutaraldehyde solution with a mass fraction of 25%, and then vacuumized until the degree of vacuum reaches 0.085Mpa, and cross-linked for 12 hours to ...

Embodiment 2

[0068] The present invention uses scanning electron microscopy (SEM), attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR), hydrogen nuclear magnetic resonance spectroscopy ( 1 H NMR), thermogravimetric (TG) and hemolysis-coagulation test, anti-protein and blood cell adhesion test, cancer cell capture and release test to characterize the various properties of the functionalized nanofibrous membrane prepared in the present invention and its combination with microfluidics. Potential application of chips in sorting circulating tumor cells.

[0069] Scanning Electron Microscopy Test:

[0070] The morphology and the diameter of the PEI / PVA nanofiber membrane that adopts SEM to characterize embodiment 1 step (1) to obtain, SEM result is as follows figure 1 As shown, the PEI / PVA nanofiber membrane prepared by electrospinning technology has a smooth surface, uniform size, and an average diameter of 365 nm.

[0071] H NMR spectrum test:

[0072] use 1 The...

Embodiment 3

[0078] Hemolysis test:

[0079] The hemocompatibility of nanofibrous membranes before and after modification was studied by hemolysis test. 1 mL of fresh healthy adult whole blood was collected, centrifuged at 2000 r / min for 5 min, and then washed three times with phosphate buffered saline (PBS) to obtain red blood cells (HRBCs). The HRBCs were diluted 30 times with PBS solution, and 0.2 mL of the HRBCs dilution was transferred to a 1.5 mL centrifuge tube containing 0.8 mL of PBS as a negative control. Transfer 0.2mL of the HRBCs dilution to a 1.5mL centrifuge tube containing 0.8mL of distilled water as a positive control. Subsequently, the configured HRBCs dilution is diluted 5 times with PBS again, and the PEI / PVA nanofiber membrane after the cross-linking treatment of Example 1 step (1), the PEI / PVA-PMPC nanofiber membrane of step (3) 4 mg of the PEI / PVA-PMPC-RGD nanofiber membranes in step (5) were immersed in 1 mL of the 5-fold diluted HRBCs suspension, and 5 parallel s...

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Abstract

The invention relates to a preparation method for a micro-fluidic chip for a zwitter-ion-embedded functionalized nanofiber membrane. The preparation method includes the steps that electrostatic spinning and steam crosslinking are carried out with a PEI/PVA spinning solution, so that a PEI/PVA nanofiber membrane is obtained; zwitter-ions MPC and a targeted ligand 4-FBA-PEG-RGD are adopted for surface modification, so that PEI/PVA-PMPC-RGD is obtained; and plasma bonding with a fishbone-shaped PDMS micro-fluidic channel cover plate is carried out, so that the micro-fluidic chip for the zwitter-ion-embedded functionalized nanofiber membrane is obtained and can be used for sorting circulating tumor cells. The main material is simple in preparation process, raw materials are cheap and easy to obtain, the sample treatment process is easy and convenient to operate, and efficient capturing and lossless releasing of circulating tumor cells in blood can be quickly achieved; and the micro-fluidicchip has excellent anti-protein and anti-hemocyte adhesion properties, improves the purity of captured cells and has good application prospects at the aspects of CTC sorting, detection and analysis.

Description

technical field [0001] The invention belongs to the technical field of microfluidic chips and cell sorting, and in particular relates to a method for preparing a microfluidic chip embedded with zwitterion-functionalized nanofiber membranes. Background technique [0002] According to the report of the World Health Organization in 2014, cancer has become the number one killer threatening human life and health. More than 90% of cancer patients die from tumor metastasis and recurrence. In the early stage of tumor metastasis, tumor cells detach from the primary solid tumor and enter the blood, becoming circulating tumor cells (CTC). Studies have shown that CTCs have been found in the blood of many tumors with a diameter of less than one millimeter [PiergaJ-Y, et al. Clinical significance of immunocytochemical detection of tumor cells using digital microscopy in peripheral blood and bone marrow of breast cancer patients. Clinical Cancer Research.2004,10:1392-1400.], and the numb...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/00C12N5/09D06M13/21D06M15/53D06M15/356D04H1/728D01D5/00D06M101/30D06M101/24
Inventor 史向阳肖云超王梦媛
Owner DONGHUA UNIV
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