CN102876564A

CN102876564A - High-flux biomaterial-screening microfluidic chip

Info

Publication number: CN102876564A
Application number: CN2012104210461A
Authority: CN
Inventors: 廖晓玲; 刘波; 徐文峰
Original assignee: Chongqing University of Science and Technology
Current assignee: Chongqing University of Science and Technology
Priority date: 2012-10-29
Filing date: 2012-10-29
Publication date: 2013-01-16

Abstract

The invention provides a high-flux biomaterial screening microfluidic chip which is characterized in that a microfluidic detection device is attached to a substrate, wherein the microfluidic detection device is provided with a cell sample introduction element from which more than 2 channels are led out; each channel is communicated with a plasmid transfection element, and the outlet of the plasmid transfection element is communicated with a serpentine tube; the outlet of each serpentine tube is divided into more than 2 channels; each channel is communicated with a cell capture element; and each cell capture element is communicated with a biochemical reagent and growth factor loading element. The invention can simultaneously detect cell integral shape and intracellular labeled protein activity variation under different biomaterial actions on one chip, and monitor the action effect of the biomaterial on the actual cells, thereby greatly lowering the detection cost, enhancing the detection accuracy and detection efficiency, and achieving the goal of biomaterial screening.

Description

A kind of high-throughput biomaterial screening micro-fluidic chip

Technical field

The present invention relates to a kind of biochip, specifically is a kind of high-throughput biomaterial screening micro-fluidic chip.

Technical background

Have the research emphasis that the biomaterial of organizing inducing function has become contemporary technical field of biological material, its core is by material self optimization design, stem cell directional differentiation in the inductor.Screen optimal biomaterial, traditional method is mainly by material and co-culture of cells, utilize cellular form variation, multiplication rate and by biochemical means analysis in a large number through the cell pyrolysis liquid of digestion process, or observe some functional proteins of the cell that is fixed by immunofluorescence technique, come indirectly the test and evaluation material on the impact of cell behavior.There is inefficiency in this method, the problems such as poor controllability, and indirectly detected result is difficult to the real cellular change of reaction.Present stage, micro-fluidic chip is exactly cytobiology in one of most important Application Areas of biology.Micro-fluidic chip research to as if viable cell, but the cellular change under the direct viewing experiment condition is that common albumen or gene chip are irreplaceable.

Therefore, need a kind of real-time detection viable cell in the micro-fluidic detection chip of the behavior of biomaterial, to screen efficiently biomaterial.

Summary of the invention

The object of the present invention is to provide a kind of simple in structure, cost is low and can realize detecting simultaneously the variation under different biomaterial effects of cell configuration and cell inner mark protein-active on the chip piece, the Real Time Monitoring biomaterial is to the action effect of true cell, improve Detection accuracy and detection efficiency, reach the purpose of biomaterial screening.

In order to address the above problem, technical scheme of the present invention is: a kind of high-throughput biomaterial screening micro-fluidic chip, comprise substrate and micro-fluidic proofing unit two portions, it is characterized in that: micro-fluidic proofing unit is attached on the substrate, micro-fluidic proofing unit has a cell sample introduction element, draw passage more than 2 by it, every channel connection a plasmid transfection element, and the outlet of plasmid transfection element communicates with snake pipe again; The passage more than 2 is told in the serpentine channel outlet, and every channel connection a cell capture element, and each cell capture element and biochemical reagents and somatomedin loading element are interconnected; The final outlet of cell capture element is connected with waste liquid pool.

In the technique scheme, described biochemical reagents and somatomedin loading element ingress are provided with the little valve of unidirectional drive.

As the preferred embodiment of technique scheme, on the substrate below the described cell capture element, being coated with in advance needs detected biomaterial.

Preferred embodiment as technique scheme, the element of described micro-fluidic proofing unit is realized by photoetching technique with device, be the PDMS material, PDMS: polydimethylsiloxane (polydimethylsiloxane) is a kind of polymer materials that is widely used in the field such as micro-fluidic.Its cost is low, uses simply, with having good adhesivity between the silicon chip.Biomaterial is by adopting mechanical manipulator accurately to be coated with to substrate the mode subregion of spraying; Substrate is made by transparent material.

Because above-mentioned cell capture element can be caught different cells, so biomaterial to be detected in its lower section, various kinds of cell just can be detected simultaneously to its consistency.Therefore simultaneously, a plurality of cell capture elements are arranged on the chip, can spray multiple biomaterial in its lower section, can also realize that the cell compatibility and the tissue that detect simultaneously multiple biomaterial on chip piece induce effect.

Main effect of the present invention is: owing to being assembled with cell sorting element, cell capture element and plasmid transfection element at same chip, in testing process, realization is detecting on the chip piece under the different biomaterial effects simultaneously, the variation of cell configuration and cell inner mark protein-active, the monitoring biomaterial is to the action effect of true cell, greatly reduce testing cost, improve Detection accuracy and detection efficiency, reach the purpose of biomaterial screening.

Description of drawings

Fig. 1 is plan structure schematic diagram of the present invention;

Fig. 2 is that the present invention is from the biomaterial spraying partial schematic diagram of bottom substrate side-looking.

Among the figure: 1. substrate; 2. micro-fluidic proofing unit; 3. cell sample introduction element; 4. passage; 5. plasmid transfection element; 6. serpentine channel; 7. the little valve of unidirectional drive; 8. biochemical reagents and somatomedin loading element; 9. cell capture element; 10. waste liquid pool; 11. biomaterial; 12. the another kind of biomaterial different from 11.

Embodiment

The present invention is further illustrated below in conjunction with accompanying drawing.

Referring to Fig. 1 to Fig. 2: shown in a kind of high-throughput biomaterial screening micro-fluidic chip, the

different biomaterials

11,12 that at first will need to screen adopt mechanical manipulators, and accurately the mode subregion of spraying is coated to chip substrate 1.Join on the chip estimating stem cell that biomaterial the uses cell sample introduction element 3 by micro-fluidic proofing unit 2; Cell enters plasmid transfection element 5 through passage 4, induces the biological probe transfection of effect to enter in the stem cell with estimating stem cell, allows cell be arranged as single linear stream through serpentine channel 6 again, enters cell capture element 9 and catches individual cells; Add the Growth of Cells nutritive medium with the little valve 7 of unidirectional drive by biochemical reagents and somatomedin loading element 8, make it adhere to growth, hatch enough time.High-throughput biomaterial screening micro-fluidic chip is placed under the microscope inspection examining system, gathers different wave length fluorescence in the cell with fluorescence CCD, automatically calculate fluorescence efficiency.Detect the activity change of specific protein in the cell according to the fluorescence efficiency of different probe, determine the practical function effect that different biomaterials are induced stem cell.Last waste liquid flows into waste liquid pool 10 collection and treatments.

Claims

1. a high-throughput biomaterial screens micro-fluidic chip, comprise substrate (1) and micro-fluidic proofing unit (2) two portions, it is characterized in that: micro-fluidic proofing unit (2) is attached on the substrate (1), micro-fluidic proofing unit (2) has a cell sample introduction element (3), draw passage (4) more than 2 by it, every passage (4) is being communicated with a plasmid transfection element (5), and the outlet of plasmid transfection element (5) communicates with snake pipe (6) again; The passage more than 2 is told in serpentine channel (6) outlet, and every channel connection a cell capture element (9), and each cell capture element (9) and biochemical reagents and somatomedin loading element (8) are interconnected; The final outlet of cell capture element (9) is connected with waste liquid pool (10).

2. a kind of high-throughput biomaterial according to claim 1 screens micro-fluidic chip, and it is characterized in that: described biochemical reagents and somatomedin loading element (8) ingress are provided with the little valve of unidirectional drive (7).

3. a kind of high-throughput biomaterial screening micro-fluidic chip according to claim 1 is characterized in that: on the substrate (1) below the described cell capture element (9), be coated with in advance the biomaterial (11) that need to be detected.

4. a kind of high-throughput biomaterial according to claim 1 screens micro-fluidic chip, it is characterized in that: the element of described micro-fluidic proofing unit (2) is realized by photoetching technique with device, is the PDMS material; Biomaterial (11) is by adopting mechanical manipulator accurately to be coated with to substrate (1) the mode subregion of spraying; Substrate (1) is made by transparent material.