High-throughput screening apparatus using droplet micro-fluidic chip

A technology of microfluidic chips and screening equipment, applied in the direction of microorganism-based methods, biochemical equipment and methods, specific-purpose bioreactors/fermenters, etc. Droplet screening solutions and system integration need to be improved to achieve the effects of saving droplet samples, simple droplet supply, and low processing and manufacturing costs

Pending Publication Date: 2018-05-11
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Abstract
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Problems solved by technology

However, all kinds of high-throughput screening equipment that have been put into use are mainly large-scale equipment systems that are expensive and complex to operate developed by foreign companies. In particular, flow cytometers are required as the main component to meet high-throughput requirements. The flow cytometer provided by the company is expensive and requires special personnel to operate. The screened object needs to have fluorescent signals on the surface or inside, and cannot detect and screen metabolites secreted by cells, which limits its application range.
[0003] The Chinese invention patent "A System and Method for Fluorescence Detection of Droplet Samples" (authorized announcement number CN104388307A) discloses a system and method for fluores

Method used

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  • High-throughput screening apparatus using droplet micro-fluidic chip
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  • High-throughput screening apparatus using droplet micro-fluidic chip

Examples

Experimental program
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Effect test

Embodiment

[0116] Example: Fluorescent detection of Bacillus subtilis droplets expressing red fluorescent protein

[0117] Pick a single colony from a fresh Bacillus subtilis LB solid plate expressing red fluorescent protein to LB liquid medium, culture at 37°C and 200rpm until OD600=0.6-0.8, wash the cells with PBS buffer at pH 7.4 3 times, 775g each time, 5min, and finally resuspend the bacteria with PBS buffer, dilute the bacteria to OD600=0.125, select the cell embedding chip for droplet embedding of cells, set the flow rate of the oil phase to 200 μL / h, The flow rate of the Bacillus subtilis cell diluent was 50 μL / h, and the generated droplets were injected into a 1mL syringe. The ratio of burying two or more cell droplets is 0.0902.

[0118] The droplet detection chip 2 is placed on the stage 37, and the position of the stage 37 is adjusted so that the droplet detection chip 2 is clearly imaged on the high-speed camera. Set the droplet flow rate to 10 μL / h, the oil phase flow rat...

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Abstract

The invention provides a high-throughput screening apparatus using a droplet micro-fluidic chip. The apparatus comprises: a rack; a micro-fluidic control system constructed to control a fluid supply device for supplying a droplet-containing fluid to the droplet micro-fluidic chip; a photoelectric detection system comprising an incident light generation device for limiting outgoing and incident light paths, a preferable laser excitation device and a fluorescence detection device for limiting a fluorescent light path; an electric screening system constructed to connect the droplet micro-fluidicchip in order to screen droplets flowing through the droplet micro-fluidic chip; and one or more controllers constructed to receive a signal from the fluorescence detection device, compare the signalwith a predetermined threshold and control the electric screening system to perform droplet screening according to the obtained comparison result.

Description

technical field [0001] The invention relates to a high-throughput screening device using a droplet microfluidic chip, especially a single-cell screening device. The invention also relates to a droplet screening method using the device. Background technique [0002] Screening from nature and mutation evolution in the laboratory has always been an important way to develop and innovate industrial microbial strains, and is the foundation of the industrial biotechnology industry. The microbial strains currently used in industry are mainly obtained through mutagenesis screening. At present, the screening of microbial strains mostly adopts micro-plate method (96-well plate or 384-well plate) or plate method. These methods almost all rely on manual operation, and it takes a long time to screen more than 100,000 mutant libraries and consumes a lot of time. The cost of consumables and reagents is unaffordable. The microbial high-throughput screening platform uses advanced modern au...

Claims

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Application Information

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IPC IPC(8): G01N15/00C12M1/36C12M1/34C12R1/125
CPCC12M47/04G01N15/00G01N2015/0065G01N2015/03
Inventor 王钦宏涂然袁会领吴晟
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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