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Method and device for constructing three-dimensional microenvironment

A micro-environment and three-dimensional technology, applied in enzymology/microbiology devices, biochemical cleaning devices, biochemical equipment and methods, etc., can solve problems such as reducing the total cost of research, and achieve simple operation, flexible application, and wide application Effect

Active Publication Date: 2015-08-05
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the expensive automated operating system has not effectively reduced the total cost of research, and it is necessary to develop more efficient and simpler loading methods to achieve high-throughput three-dimensional micro-scale array chips of drugs, materials and cells

Method used

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  • Method and device for constructing three-dimensional microenvironment
  • Method and device for constructing three-dimensional microenvironment
  • Method and device for constructing three-dimensional microenvironment

Examples

Experimental program
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Effect test

preparation example Construction

[0059] The preparation of the microsponge scaffold display involves various pattern manufacturing and microfabrication techniques including photolithography, microcontact printing, microfluidic patterning, Laminar flow patterning, stencil patterning, Imprint lithography, flow lithography, etc. The above-mentioned technologies have their own advantages and disadvantages in the production process, and they can be used in combination with other technologies to achieve a better three-dimensional patterning effect. For example, in photolithography, using a digital micromirror device (digital micromirror device DMD) instead of a photomask can dynamically change the pattern during the production process, thereby producing a three-dimensional pattern with high complexity and high internal correlation. The selection of pattern manufacturing, micro-manufacturing technology and combined use technology needs to comprehensively consider the physical and chemical properties of materials and...

Embodiment 1

[0079] Embodiment 1, the manufacture of the device for constructing three-dimensional microenvironment

[0080] 1. Combining photolithography, salting-out technology and freeze-drying method to prepare microsponge scaffold display

[0081] The biological material for preparing the microsponge scaffold display provided in this example is the monomer polyethylene glycol diacrylate (PEGDA4000).

[0082] The polyethylene glycol diacrylate selected in this example has the advantages of good biocompatibility, mechanical properties, non-degradability, and controllability of the degree of photocrosslinking, and is suitable as a biomaterial for the preparation of microsponge scaffolds in this example. . The slide glass and cover glass used in the display process of preparing the microsponge holder with supporting base B are commercial slide glass and cover glass, wherein the size of the slide glass is 76.2mm×25.4mm, and the thickness of the cover glass is is 150 μm.

[0083] The fab...

Embodiment 2

[0133] Example 2. Utilizing the device provided by the present invention to realize three-dimensional microscale efficient loading of sample liquid

[0134] Utilize the device prepared in embodiment 1 (the microsponge support array is figure 2 The microsponge support array shown in B, on which the volume of the microsponge support is 0.2355μl) realizes the efficient three-dimensional microscale loading of the sample solution, and the schematic diagram of the process is shown in Figure 4 . The liquid thin-layer substrate is covered on the display of the micro-sponge bracket, and the liquid is automatically sucked into the micro-sponge bracket by using the adsorption of the micro-sponge and the easy-flowing separation of the liquid thin layer, so as to realize the three-dimensional micro-scale loading of the sample liquid. The sample solution can be any one or a mixture of small molecular compounds, drugs, nucleic acids, proteins, extracellular matrix components, polymer mate...

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Abstract

Provided are a method and a device for constructing a three-dimensional cellular microenvironment. The method comprises the following steps: 1) establishing a three-dimensional transparent sponge scaffold or a three-dimensional transparent sponge scaffold array; 2) establishing a thin sample liquid layer containing molecules, a material, cells, and a mixture thereof; 3) combining the transparent sponge scaffold or transparent sponge scaffold array in 1) with the thin liquid layer in 2), and completing loading of a sample liquid, thereby achieving construction of a three-dimensional microenvironment. The method and device provide a simple and practical platform for research on the growth, proliferation, label-free observation, and functional analysis of cells in a three-dimensional microenvironment in the fields such as the biomedical research and the drug research and development, and rapidly and losslessly realize the patterning of the three-dimensional microenvironment of molecules, a material, cells, and a mixture thereof, high flux synchronous loading, and label-free real-time observation.

Description

technical field [0001] The present invention relates to a method and device for constructing a three-dimensional microenvironment, in particular to a method and device for efficiently realizing three-dimensional microscale arrangement of drugs, materials and cells based on the automatic loading effect of a three-dimensional patterned "micro-sponge scaffold" , belonging to the technical field of three-dimensional microscale chips. Background technique [0002] The development of fields such as molecular cell biology, diagnosis and clinical medicine, and new drug development requires the study of cell-drug, cell-extracellular matrix (ECM), and cell-cell interactions in vitro. Traditional research methods are based on the two-dimensional microenvironment of commercial petri dishes or multiwell plates. This simplified two-dimensional microenvironment cannot simulate and reproduce the three-dimensional environment in the body well. Therefore, constructing a three-dimensional mi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/00C08L71/08C08J3/28C08J3/24
CPCC12N5/0062
Inventor 杜亚楠赵姗
Owner TSINGHUA UNIV
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