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Microfluidic chip for sorting and its application

A microfluidic chip, chip technology, applied in the direction of using electricity/wave energy to treat microorganisms, specific-purpose bioreactors/fermenters, microorganisms, etc., can solve the problems of repeatability to be verified, limited chip width, chip flow and Dealing with issues such as low sample capacity

Inactive Publication Date: 2016-06-29
SHANGHAI EAST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the manufacturing process of this type of chip is relatively complicated, and the experimental condition control requirements are high, so the repeatability of the experiment has yet to be verified.
In 2012, Kang et al. reported a magnetic separation microfluidic chip based on a dual-channel mode. A small chamber for collecting CTCs was provided on the same side wall of two parallel microchannels, and a permanent magnet was placed outside the end of the small chamber for the microfluidic separation. A sufficiently strong magnetic field is generated in the channel. When the tumor cells labeled with magnetic beads flow through the microchannel, they will be deflected perpendicular to the direction of the flow velocity under the action of the magnetic field force and enter the collection chamber on the side wall, thereby realizing the separation of CTCs. However, the main channel width of the chip is limited, so the flow rate and sample processing capacity of the chip are low
However, the main disadvantage of this chip is that the immunomagnetic bead-labeled tumor cells attached to the cover glass by the force of the magnetic field are still subjected to strong fluid disturbance and exposed to fluid shear stress, resulting in the detachment and loss of some of the attached cells.

Method used

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  • Microfluidic chip for sorting and its application
  • Microfluidic chip for sorting and its application
  • Microfluidic chip for sorting and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 3

[0144] Example 3 Sorting of circulating tumor cells in the peripheral blood of patients with colon cancer using the chip of the present invention

[0145] (1) Take 5ml of peripheral venous blood from patients with colon cancer, and remove the first 2ml of peripheral blood to avoid skin epithelial cell contamination. The remaining blood samples are collected in EDTA anticoagulant tubes, stored and transported at 4 degrees Celsius, and tested within 4 hours.

[0146] (2) Flush the channel of the microfluidic chip with block polyether F-108 for 30 minutes to prevent non-specific cell adhesion, and then place a permanent magnet with a magnetic field strength of 500mT above the capture area of ​​the microfluidic chip.

[0147] (3) According to 10 7For the ratio of magnetic beads / ml blood, add 150 microliters of immunomagnetic beads conjugated with anti-human epithelial cell adhesion molecule antibody (purchased from Invitrogen) to 3 ml of blood in step (1), and incubate at 4°C 30 ...

Embodiment 4

[0153] Example 4 Sorting of CD133 colon cancer tumor stem cells using the chip of the present invention

[0154] (1) Flush the channel of the microfluidic chip with block polyether F-108 for 30 minutes in advance to prevent non-specific cell adhesion, and then place a permanent magnet with a magnetic field strength of 500mT above the capture area of ​​the microfluidic chip.

[0155] (2) Adjust the single cell suspension sample to 1X10 8 cells / 300 microliters, according to 100 microliters / 10 8 Add blocking antibody to the proportion of cells to block FcR receptors.

[0156] (3) According to each 1X10 8 Add 100 microliters of anti-human CD133 immunomagnetic beads (purchased from Miltenyi Biotec) to the cells, add the immunomagnetic beads to the reaction solution in step (2), mix well, and incubate at 4°C for 30 minutes.

[0157] (4) Move all the fully reacted samples in step (3) to the micro-syringe pump, and continuously pump the samples into the main channel of the microflu...

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Abstract

The invention relates to a microfluidic chip for sorting and an application thereof, and in particular discloses a microfluidic chip for sorting. The chip comprises a chip inlet, a main channel and a chip outlet, wherein the chip inlet is used for a liquid to be sorted to flow in; the main channel is used for the liquid to be sorted to flow; the chip outlet is used for discharging the sorted liquid; the main channel includes an inlet channel, an outlet channel and a capturing area channel, wherein the inlet channel is positioned on the chip inlet, the outlet channel is positioned on the chip outlet, and the capturing area channel is positioned between the inlet channel and the outlet channel; and a plurality of small collecting rooms which are provided with downward openings and used for collecting target materials to be sorted are formed in the capturing area channel. In addition, the invention further provides a microfluidic chip device and a microfluidic chip kit for sorting and a method for sorting target cells or other substances. The microfluidic chip disclosed by the invention has the advantages of simplicity in manufacturing and good operability and repeatability, the area of a cell sorting area is greatly increased, and the high-flow cell sorting is supported.

Description

technical field [0001] The invention relates to the technical field of microfluidic chips, in particular to a sorting microfluidic chip, a microfluidic chip device, a microfluidic chip kit, and a sorting method for sorting objects. Background technique [0002] Microfluidic chip technology, also known as lab-on-a-chip (lab-on-a-chip), is a micro-machining technology proposed by Swiss researchers in the 1990s to separate and sample components involved in biochemical reactions. The basic steps such as, reaction and detection are integrated on a chip with an area of ​​about several square centimeters, and the whole process is precisely controlled. The microchannels inside the chip form a controllable microfluidic network, which can be used to replace conventional biochemical detection, and has shown great development in the fields of medicine and life sciences with its characteristics of miniaturization, integration, high throughput and high precision potential and application...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/00C12M1/42C12N5/00C12N5/09C12N13/00C12N15/10C07K1/14
Inventor 张振宇葛海燕印晓伟代镇岭李书恒李淑萍
Owner SHANGHAI EAST HOSPITAL