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Semi-contact under-oil continuous droplet sample applying and liquid adding method

A semi-contact and droplet technology, which is applied to the analysis of materials and instruments, can solve problems such as cross-contamination and false-positive screening results, and achieve the effects of increasing speed, improving reliability and uniformity, and facilitating analysis and screening throughput

Active Publication Date: 2014-07-30
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, since the capillary tip is frequently in contact with droplets of different samples, it is easy to cause cross-contamination, resulting in false positive screening results

Method used

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  • Semi-contact under-oil continuous droplet sample applying and liquid adding method
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  • Semi-contact under-oil continuous droplet sample applying and liquid adding method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] figure 1 It is a schematic diagram of the principle of using the semi-contact liquid droplet continuous spotting and liquid addition method to generate sample droplets and add reagents. The specific operation process of generating sample droplets 6 by continuous pointing of droplets ( figure 1 (1)-(4)) are as follows: the sampling end of capillary 1 is sharpened to reduce cross-contamination during sampling, and the inner wall of capillary 1 is treated with hydrophobic surface to prevent the adsorption of samples and reagents on its surface . A layer of oil phase 4 immiscible with water is covered on the microwell array chip 2 . Aspirate a certain volume of sample solution 5 to be sampled into the capillary 1 , and move the translation stage so that the capillary 1 is placed directly above the center of the microwell 3 of the microwell array chip 2 . According to the volume of the sample droplet, the distance d1 between the tip of the capillary 1 and the lower surfac...

Embodiment 2

[0055] figure 2 It is a schematic diagram of the principle of the reagent droplet generation and sample addition operation using the semi-contact liquid droplet continuous pointing and liquid addition method. The specific operation process of generating reagent droplets 8 by continuous pointing of droplets ( figure 2 (1)-(4)) are as follows: the sampling end of capillary 1 is sharpened to reduce cross-contamination during sampling, and the inner wall of capillary 1 is treated with hydrophobic surface to prevent the adsorption of samples and reagents on its surface . A layer of oil phase 4 immiscible with water is covered on the microwell array chip 2 . Aspirate a certain volume of reagent solution 7 to be sampled into the capillary 1 , and move the translation stage so that the capillary 1 is placed directly above the center of the microwell 3 of the microwell array chip 2 . According to the volume of the spotting liquid droplet, the distance d1 between the tip of the cap...

Embodiment 3

[0058] image 3 The picture of the fluorescein droplet array generated by the continuous droplet spotting method, and the picture of the droplet array obtained by diluting the fluorescein droplet array with equal volume by the continuous liquid addition method. In the experiment, the inner diameter of the capillary is 150 microns, and the outer diameter is 200 microns. The sampling end is sharpened, and the size of the tip is about 30 microns. The inner wall and tip of the capillary were hydrophobized with octadecyltrichlorosilane. Use a syringe pump equipped with a 1 microliter syringe as the liquid drive system, and connect the syringe to the capillary. The microhole array is processed on the glass substrate by photolithography and wet etching, the size of each microhole is 260 microns in diameter, 60 microns in depth, and the distance between the centers of the microholes is 400 microns. Octadecyltrichlorosilane was also used to treat the surface of the microwell array c...

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Abstract

The invention provides a semi-contact under-oil continuous droplet sample applying and liquid adding method, which is suitable for sequentially operating a droplet array system. According to the method, the distance between the pointed end of a capillary tube sample applying needle and the lower surface of a micro-hole (or a generated droplet) is controlled accurately, and the affinity or interface tension interaction between the droplet and the surface (or the generated droplet) is utilized, so that rapid and reliable continuous droplet sample applying or continuous liquid adding is realized, and the problem of cross infection during sample applying is solved effectively. The method is suitable for biochemical analysis screening researches such as high-flux medicament screening, protein crystallization condition screening, enzyme kinetics research and drug toxicity determination.

Description

technical field [0001] The field of the invention relates to the field of liquid droplet analysis, in particular to a semi-contact method for continuously spotting and adding liquid to liquid droplets under oil. Background technique [0002] Droplet-based microfluidics has achieved rapid development and wide application. The main advantages of droplet microfluidics include: 1) The volume of the droplet reactor can be flexibly adjusted from femtoliter to nanoliter, so the consumption of samples and reagents can be reduced to an extremely low level; 2) It can be generated in a short time A large number of droplet reactors can significantly improve the throughput of analysis and screening; 3) Rapid mass transfer, heat transfer and mixing of components can be realized in the droplets, which effectively improves the efficiency of biochemical reactions; 4) The encapsulation of the oil phase, Effectively reduce the evaporation, diffusion and cross-contamination of trace biochemica...

Claims

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Application Information

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IPC IPC(8): G01N35/10
Inventor 祝莹方群张云霞朱丽娜
Owner ZHEJIANG UNIV
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