High-throughput micro-droplet serial dilution device and high-throughput micro-droplet serial dilution method

A gradient dilution and micro-droplet technology, applied in chemical instruments and methods, mixing methods, transportation and packaging, etc.

Pending Publication Date: 2018-11-20
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0013] In view of the above-mentioned defects of the prior art, the technical problem to be solved by the present invention is to carry out accurate gradient dilution to nanoliter levels and smaller volumes, high-throughput (multi-channel) parallel operation, and simultaneously perform multiple ratio parallel operations. Operation, without complex labor or expensive pipetting robots, convenient to control the environmental (temperature) conditions of gradient dilution

Method used

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  • High-throughput micro-droplet serial dilution device and high-throughput micro-droplet serial dilution method
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  • High-throughput micro-droplet serial dilution device and high-throughput micro-droplet serial dilution method

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Embodiment approach 1

[0071] Specific embodiment one, using the above-mentioned high-throughput micro-droplet gradient dilution device to dilute the solution containing the dye or fluorescent dye, including the following steps:

[0072] 1) if Figure 11 As shown, in the initial position, an aqueous solution containing dye (concentration C1) is injected through series 1 fluid channel 3 by using positive pressure (such as positive air pressure, etc.) at the liquid inlet port or negative pressure (such as vacuum, etc.) at the liquid outlet port Into the chip, and the aqueous solution without dye is injected into the chip through the series 2 fluid channel 4;

[0073] 2) The positions of the upper sub-chip and the lower sub-chip are relatively moved, so that series 1 microwell 1 (volume V1) and series 2 microwell 2 (volume V2) are no longer partly superimposed on each other, thereby forming independent microdroplets

[0074] 3) if Figure 6 As shown, the upper sub-chip and the lower sub-chip are rela...

specific Embodiment approach 2

[0077] The second specific embodiment is to dilute the gene as a digital gene amplification method in a large linear space. Digital nucleic acid amplification is a new method for precise quantification of gene molecules. It distributes a reaction solution containing a specific concentration of the target gene into a large number of micro-droplets, so that some of the micro-droplets contain a single molecule of the target gene and the remaining micro-droplets do not contain the target gene. Through the gene amplification reaction (this amplification reaction can be a PCR reaction, can also be isothermal gene amplification or other gene amplification methods), the single-molecule target gene in the micro-droplet is amplified in large quantities. Correspondingly, if gene fluorescent dyes or molecular probes are used, the fluorescence intensity can be enhanced due to the increase of the concentration of the target gene. In this way, the original micro-droplet containing the targe...

specific Embodiment approach 3

[0080] The third specific embodiment is to dilute the medicine and analyze the effectiveness of the medicine. In the process of drug development, it is usually necessary to study the inhibitory or killing effect of the drug on target cells (such as cancer cells or bacteria) and find its semi-lethal dose IC50; it is also usually necessary to study the effect of drugs on normal cells (such as white blood cells) toxicity. The usual method is to manually pipette or use a pipetting robot to generate a drug gradient solution such as 100%, 50%, 25%, 12.5%...etc. in cell culture plates such as 96-well plates or 384-well plates, and Put target cells or normal cells to study their response to specific concentrations of drugs. The chip of the present invention can easily realize this process. The volume V1 of microwell 1 of series 1 and the volume V2 of microwell 2 of series 2 are 1:1, the drug solution with 100% concentration is injected into the fluid channel 3 of series 1 at the ini...

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Abstract

The invention discloses a high-throughput micro-droplet serial dilution device and a high-throughput micro-droplet serial dilution method and relates to the field of serial dilution. The high-throughput micro-droplet serial dilution device comprises an upper subchip and a lower subchip, wherein relative displacement of the upper subchip and the lower subchip reaches a second position; micropores of series 1 are superposed with those of a first-row series 2; the concentration of a solution 1 in the micropores of the series 1 is changed by a solution 2 in the micropores in the first-row series 2; after the solutions are sufficiently mixed, the upper subchip is displaced to the third position relative to the lower subchip, and the micropores of the series 1 are enabled to be superposed with micropores in a second-row series 2; and the concentration of the solution 1 in the morepores of the series 1 is changed by the solution 2 in the micropores of the series 2 for the second time. Multiple relative displacement of the upper subchip and the lower subchip can be realized, and thereby the aim of performing gradient change of the solution 1 in the micropores of the series 1 is achieved. The high-throughput micro-droplet serial dilution device and the high-throughput micro-droplet serial dilution method disclosed by the invention can perform accurate serial dilution for nano-liter level or smaller volume and multi-proportion parallel operation at the same time; and complex workers or expensive liquid transfer robots do not be needed and environmental conditions of serial dilution are conveniently controlled.

Description

technical field [0001] The invention relates to the field of gradient dilution, in particular to a high-throughput micro-droplet gradient dilution device and method. Background technique [0002] Serial dilution has been widely used in the fields of chemistry, biology and medicine for a long time. For example, to test the effectiveness of a new compound against bacteria, the drug is usually tested by gradient dilution (for example, concentration 100nM, 10nM, 1nM, 100pM...). [0003] The general method of gradient dilution is to dilute the initial solution containing a specific concentration of the target substance with a diluent in a certain proportion, and then take the diluted solution and the diluent to dilute again in the same or different proportions. The same steps can be Repeat the operation until the concentration of the target substance that needs to be diluted is reached. For example, if you need to prepare 1nM NaOH solution from 1mM NaOH solution, you can use a ...

Claims

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Application Information

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IPC IPC(8): B01F3/08B01L3/00
CPCB01L3/00B01F23/405B01F23/40B01F2101/23
Inventor 沈峰俞梦超
Owner SHANGHAI JIAO TONG UNIV
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