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Automatic micro-droplet array screening system using method with pico-liter-scale precision

A micro-droplet and drive system technology, applied in chemical instruments and methods, laboratory equipment, laboratory containers, etc., can solve problems such as difficult to use screening systems, difficult to industrialize instruments, difficult to realize automation, etc., to achieve improved Throughput, reduced risk of cross-contamination, effect on reduced sample/reagent consumption

Active Publication Date: 2013-04-03
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, such methods are complex in structure, difficult to process, and difficult to use in screening systems with a large number of different samples
Moreover, the implementation of the above two types of methods requires complex manual adjustment methods such as liquid flow rate adjustment, droplet frequency, droplet size control, droplet position feedback, etc., which is difficult to achieve reliable automation, and there is a big problem in the industrialization of instruments. Difficulties

Method used

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  • Automatic micro-droplet array screening system using method with pico-liter-scale precision
  • Automatic micro-droplet array screening system using method with pico-liter-scale precision
  • Automatic micro-droplet array screening system using method with pico-liter-scale precision

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Experimental program
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Effect test

Embodiment 1

[0058] image 3 is based on figure 1 and figure 2 The liquid droplet array screening system and its use method use 32 small molecule chemicals as the samples to be screened, and Caspase-1 enzyme as the screening target to carry out the fluorescence pictures obtained by screening enzyme inhibitors. Start by loading 100 µM of small molecule compounds into sample / reagent storage tubes. use figure 1 The droplet array generation method generates 32 droplets of small molecule chemicals on the microwell array chip, and the volume of each droplet is 180 picoliters. then follow figure 2 In the target reagent injection method, 180 picoliters of Caspase-1 enzyme solution (6 mU / μL) and 180 picoliters of substrate (Z-YVAD-R110) solution (20 μM) were injected into each droplet to trigger a reaction. The microwell array chip was incubated at 35 degrees Celsius for 1 hour, and the experimental results were obtained by fluorescence imaging. Analyzing the experimental results, the stro...

Embodiment 2

[0061] Figure 4 is true image 3 The result recording chart of Caspase-1 enzyme half-inhibition concentration determination of No. 28 compound obtained in the screening experiment. First, fill the sample / reagent storage tubes with compound No. 28 solutions at concentrations of 0.1 nM, 1 nM, 10 nM, 100 nM, 1 μM, 10 μM, and 100 μM, respectively. according to figure 1 and figure 2In the droplet array screening system and method of use, droplets (180 picoliters) containing different concentrations of compound No. 28 solutions are generated on the microwell array chip, and then 180 picoliters of Caspase-1 is injected into each droplet Enzyme solution (6 mU / μL) and 180 pL of substrate (Z-YVAD-R110) solution (20 μM) were used to trigger the reaction. According to the method of Example 1, the resultant fluorescence pictures were obtained, and the fluorescence brightness values ​​were extracted and normalized. The concentration of compound 28 was logarithmically processed, and t...

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Abstract

The invention relates to the field of high-throughput screening and particularly relates to an automatic micro-droplet array screening system using method with pico-liter-scale precision. The using method provided by the invention comprises the following steps of: firstly, using liquid with a low thermal expansion coefficient and filled in a liquid driving system and a capillary tube as carrier liquid, and entirely emptying bubbles inside the capillary tube; secondly, soaking a sampling end of the capillary tube in an oil phase sample immiscible with a water phase sample, extracting a section of oil phase sample into the capillary tube, so as to isolate the water phase sample and the carrier liquid; thirdly, soaking the sampling end of the capillary tube into a sample / reagent storage tube and extracting a certain volume of the water phase sample solution into the capillary tube; and finally, moving the sampling end of the capillary tube into the oil phase sample above micro-pores of a micro-porous array chip, and pushing the sample solution in the capillary tube into the micro-pores, so as to form sample droplets. With the adoption of the using method provided by the invention, the liquid is quantitatively measured, and the generated droplets have the pico-liter-scale volume precision, so that the consumption of the samples / reagents in the high-throughput screening can be effectively reduced, and the experiment cost can be saved.

Description

technical field [0001] The field of the invention is the field of high-throughput screening, in particular to a method for using an automatic micro-droplet array screening system with picoliter precision. Background technique [0002] The high-throughput screening system originated from the research of drug screening. It mainly uses 96 or 384-well plates as arrayed reactors, and uses automated robots for liquid distribution and sample mixing. It uses highly sensitive and high-speed detection devices and data processing software to process The experimental results are analyzed and processed to achieve a screening throughput of at least 10,000 samples per day. Due to its powerful screening and analysis functions, the application range of high-throughput screening technology has expanded from drug screening to multiple scientific fields such as biology, medicine, and chemistry. However, with the rapid increase of new targets and sample volume, commercial high-throughput liquid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01L3/00
CPCB01L2200/0673B01L3/00B01L3/021B01L2200/0642B01L3/50853G01N35/1016G01N35/00G01N2035/1034B01L2200/142B01L3/502715B01L3/50273B01L2200/0605B01L2200/0684B01L2200/14B01L2300/0819B01L2300/0896B01L2400/0406B01L2400/0478G01N35/00029G01N2035/00158G01N2035/1039
Inventor 方群祝莹张云霞
Owner ZHEJIANG UNIV
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