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A device for forming and screening two-dimensional droplet arrays and methods of use thereof

A droplet and array technology, applied in the direction of analyzing materials, instruments, etc., can solve the problems of slow droplet generation, difficulty in spreading droplets, and complicated operation of droplet arrays, and achieve the effect of automatic detection and automatic suction.

Active Publication Date: 2018-05-29
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The experimental operation of the droplet array formed by this method is complicated. Because the micropit structure is closed at the bottom, it is difficult to spread the droplets, and the droplet generation speed is slow, generally 0.1 to 1Hz, which limits the application of this method in high-throughput screening. application

Method used

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  • A device for forming and screening two-dimensional droplet arrays and methods of use thereof
  • A device for forming and screening two-dimensional droplet arrays and methods of use thereof
  • A device for forming and screening two-dimensional droplet arrays and methods of use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Utilize the standard soft lithography technique to process on the PDMS chip 1 the micro-column 5 structure arranged in an array with a rhombic cross-section. The groove unit 6 has a diameter of 47 μm. ( Figure 12 A)

[0050] Using the green pigment solution as the droplet phase and mineral oil containing 3% ABIL EM90 surfactant as the spacer phase, a green droplet 7 with a diameter of about 38 μm was prepared using a chip with a T-shaped channel; then the sodium fluorescein solution was used as the liquid For the drop phase, mineral oil containing 3% ABIL EM90 surfactant was used as the spacer phase, and sodium fluorescein droplets 7 with a diameter of 38 μm were prepared using a chip with a T-shaped channel.

[0051] Mix two kinds of droplet suspensions 9 and pour them on the micropillar array chip 1; use the polishing sheet as the scraper 2 to make the droplet suspension 9 and the upper surface of the chip 1 generate horizontal relative motion, so as to promote the...

Embodiment 2

[0054] On the PDMS chip 1, a micro-column 5 structure arranged in an array with a square cross-section is processed by a micro-machining method. The side length of the cross-section of the micro-column 5 is 36 μm, and the gap between the micro-columns 5 is 16 μm. The micro-groove unit 47 µm in diameter ( Figure 13 A).

[0055] Using the green pigment solution as the droplet phase and mineral oil containing 3% ABIL EM90 surfactant as the spacer phase, a green droplet with a diameter of 38 μm was prepared using a chip with a T-shaped channel; then the sodium fluorescein solution was used as the droplet phase , Mineral oil containing 3% ABILEM90 surfactant was used as the spacer phase, and a chip with a T-shaped channel was used to prepare sodium fluorescein droplets with a diameter of 38 μm.

[0056] Mix the two kinds of droplet suspension 9 and pour it on the microcolumn array chip 1; use the polished sheet as the scraper 2 to make the droplet suspension 9 and the upper surfa...

Embodiment 3

[0058] Using the droplet array chip in Example 1, using the PCR reaction solution as the droplet phase, Bio-Rad's generation oil as the spacer phase, and using the T-shaped channel to prepare digital PCR droplets with a diameter of 38 μm. Place the PCR droplets in a PCR instrument for amplification. After the reaction is complete, pour the droplet suspension onto the microcolumn array chip 1; use the polished sheet as the scraper 2 to make the droplet suspension 9 move horizontally relative to the surface of the chip 1, and promote the rapid spreading of the droplet on the chip surface, forming A single-layer droplet 7 array; a spacer phase 8 oil phase is added to the surface of the micropillar array chip 1 forming the droplet array to prevent the droplet 7 from evaporating; the droplet 7 on the micropillar array chip 1 is detected. It takes about 50 seconds to spread a micropillar array chip with an area of ​​3cm×3cm, and the efficiency of trapping PCR reaction liquid droplet...

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Abstract

The present invention discloses an apparatus for forming and screening a large-scale single-layer two-dimensional liquid droplet array, and a use method thereof. The apparatus comprises a micro-column array chip, a scraping plate for auxiliary spreading of a liquid droplet, a probe for controlling the liquid droplet, a three-dimensional translation table and other components, wherein a micro groove unit for trapping the single liquid droplet is formed by using the gap between micro-columns, and the liquid droplet is spread in the micro groove unit in a single-layer and single manner through gravity effect and capillary effect or through an auxiliary means so as to form the single-layer liquid droplet array. With the apparatus of the present invention, the accurate positioning and detection on the liquid droplet, the sucking on the target liquid droplet and the subsequent treatments can be performed. According to the present invention, the apparatus has characteristics of simple structure and easy operation; and the method has advantages of fast liquid droplet spreading, high flux, high liquid droplet trapping efficiency, liquid droplet position fixing and the like, and is suitable for single cell analysis and screening, single molecule analysis and screening, high-throughput gene screening, protein directed evolution, antibody screening, microbial research, drug screening, and other fields.

Description

technical field [0001] The field of the invention is the field of microfluidic liquid droplets, in particular to a device for forming and screening a large-scale two-dimensional droplet array and a method for using the same. Background technique [0002] Droplet-based multiphase microfluidic technology is one of the hotspots in the field of high-throughput screening research in recent years. It realizes the generation, mixing, reaction, and analysis and screening of large quantities of water-in-oil or oil-in-water droplet microreactors through the control of multiphase fluids in micron-scale channels (or microstructures). Droplet-based multiphase microfluidic technology solves the problems of slow mixing speed, low analysis throughput, and easy cross-contamination in single-phase microfluidic systems, and further reduces the consumption of reagents and samples. In addition, micro-droplet technology has the advantages of automated operation, high generation frequency, and fa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N35/00
CPCG01N35/00029G01N2035/00158
Inventor 许锦钢方群祝莹黄梦诗
Owner ZHEJIANG UNIV
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