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Microfluidic chip apparatus and application thereof

A microfluidic chip and chamber technology, applied in the direction of microorganisms, laboratory containers, microorganism measurement/inspection, etc., to achieve the effect of simple production, good biocompatibility and easy operation

Active Publication Date: 2012-10-17
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can be seen that the three-dimensional culture microfluidic chip is a good in vitro experimental platform for drug screening and toxicity detection, which is expected to replace animal experiments. However, this platform has not yet been applied to the study of quantum dot cytotoxicity.

Method used

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  • Microfluidic chip apparatus and application thereof
  • Microfluidic chip apparatus and application thereof
  • Microfluidic chip apparatus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: the structure of the chip

[0042] Such as figure 1 As shown, the microfluidic chip device of the present invention is composed of a main channel and a cell culture chamber. Among them, the main channel can simulate blood vessels for transporting cell culture medium and quantum dot solution, and the cell culture chamber can simulate the adjacent tissue around blood vessels where quantum dots act.

[0043] The device includes two parts: a main channel and a cell culture chamber, and the main channel communicates with the cell culture chamber. The heights of the main channel and the cell culture chamber of the microfluidic chip device are different, 71 μm and 38 μm, respectively. In this way, the surface tension effect can be used to prevent the mixture of cells and three-dimensional culture substrate from leaking into the main channel when injected into the cell culture chamber. The cell culture chamber of the microfluidic chip device is divided into two...

Embodiment 2

[0044] Example 2: Modeling Diffusion in Tissue

[0045] Diffusion in tissues using sodium fluorescein and fluorescein isothiocyanate-labeled bovine serum albumin (FITC-BSA) as model drugs. Inject agarose solution with a mass volume ratio of 0.5% into the cell culture chambers on both sides. After it coagulates, inject 100 μM sodium fluorescein solution and 2.75 mg / mL FITC-BSA solution from the main channel respectively. At the same position on the top of the chamber, a fluorescent microscope (Leica DMI 4000B) was used to take pictures of the fluorescence intensity of sodium fluorescein every 5 minutes, and the fluorescence intensity of FITC-BSA every 10 minutes. The experiment of fluorescein sodium lasted for 120 minutes, and the experiment of FITC-BSA lasted for 300 minutes. The results are as follows figure 2 and image 3 shown. From Figure 2-4 It can be seen that the fluorescence intensity becomes stronger with time, indicating that the two substances have indeed diff...

Embodiment 3

[0046] Example 3: Three-dimensional culture of HepG2 cells in a chip

[0047] Two plates 60cm 2 The overgrown cells in the culture dish were digested, the cell suspension was centrifuged, and the supernatant was removed, and the remaining cells were redispersed to a cell density of 10 6 / mL. The three-dimensional culture matrix was prepared by mixing 100 μL 3% (w / v) low melting point agarose solution, 100 μL fetal bovine serum and 100 μL phosphate buffer solution. Equal volumes of cell suspension and three-dimensional culture matrix are mixed and injected into the cell culture chamber of the microfluidic chip of the present invention, and the chip is placed at 4°C for 10 minutes to accelerate the solidification of the agarose and wrap the cells in it. Finally, inject the culture medium into the main channel and coat the surface of the chip to prevent the volatilization of the culture medium in the channel. The chip was placed in a cell incubator, the medium was changed ever...

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Abstract

The invention provides a three-dimensional cultivation microfluidic chip apparatus used for quantum dot cytotoxicity detection. The microfluidic chip apparatus is manufactured by bonding a dimethyl silicone polymer chip and a glass substrate. The chip has two parts, which are a main channel and cell cultivation chambers. The main channel is used for simulating a blood vessel. The cell cultivation chambers communicate with the main channel with different distances, and are used for simulating adjacent tissues with different distances from the blood vessel. The heights of the main channel and the cell cultivation chambers are different, such that a mixture of cells and a three-dimensional cultivation substrate is prevented from leaking to the main channel when injected into the cell cultivation chambers. The apparatus can further be used for proofing that one of the quantum dot cytotoxicity cell mechanisms is the cell autophagia effect. The microfluidic chip apparatus provided by the invention is advantaged in simple manufacturing and easy operation. The apparatus can be used for simulating the diffusion process of the blood vessel and the adjacent tissues. With the apparatus, three-dimensional cultivation and quantum dot cytotoxicity detection of cells can be realized. Therefore, the apparatus is suitable for medicine screening and environmental toxicology analysis.

Description

technical field [0001] The invention relates to a microfluidic chip, in particular to a three-dimensional culture microfluidic chip device used for the detection of quantum dot cytotoxicity, and its application in the detection of quantum dot cytotoxicity. Background technique [0002] As an emerging nanomaterial, quantum dots have great application value in biological imaging, but their cytotoxicity limits their widespread use. Many scientists have given different explanations for the toxicity mechanism of quantum dots, such as the release of heavy metal ions, the generation of active oxygen species, and different surface properties. [0003] At present, most studies on the cytotoxicity of quantum dots use cultured cells in petri dishes or animal experiments. Documents Li Y.;Zhou Y.;Wang H.;Perrett S.;Zhao Y.;Tang Z.;Nie G.Angew Chem Int Edit,2011,50,5860-5864 proved by HepG2 cells cultured in culture dishes CdTe quantum dots coated with different chiral glutathione cause...

Claims

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Application Information

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IPC IPC(8): B01L3/00C12N5/00C12Q1/18C12Q1/02
Inventor 林金明吴静李海芳
Owner TSINGHUA UNIV
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