阅读说明：本技术 一种对细胞多角度力学测量的微流控芯片及其制作方法 (A kind of micro-fluidic chip and preparation method thereof of pair of cell multi-angle mechanical meaurement ) 是由 杨浩 朱博韬 程亮 李相鹏 李婉婷 孙研珺 于 2019-08-29 设计创作，主要内容包括：本发明公开了一种对细胞多角度力学测量的微流控芯片及其制作方法,所述的微流控芯片由设有微通道的PDMS盖片与刻蚀有电极的ITO玻璃基片键合封装而成,可在非接触条件下对细胞进行多角度力学测量,具有成本低、测量快速、对细胞损害小等优点；其制作方法简单,生产效率高,可以实现批量生产,具有广阔的商业应用前景。(The invention discloses the micro-fluidic chips and preparation method thereof of a kind of pair of cell multi-angle mechanical meaurement, the micro-fluidic chip is formed by the PDMS cover plate for being equipped with microchannel with the ito glass substrate bonding packaging for being etched with electrode, multi-angle mechanical meaurement can be carried out to cell under noncontact condition, have many advantages, such as that at low cost, measurement is quick, small to cell damage；Its production method is simple, high production efficiency, may be implemented to produce in batches, has wide commercial application prospect.)

1. the micro-fluidic chip of a kind of pair of cell multi-angle mechanical meaurement, which is characterized in that it is encapsulated by substrate and cover plate, The cover plate is the PDMS substrate equipped with microchannel, wherein the microchannel AB sections of AD section for being located relatively at center line with AC sections is right Claim setting, AE section are AD section of straight line extension, AB sections and AC sections of width having the same, AD sections and AE sections of width having the same Spend and be wider than AB sections and AC sections, the end B, C-terminal, the end D and the end E of microchannel respectively correspond the first inlet of setting, the second inlet, Third inlet and liquid outlet, ∠ BAC are 30 ° -60 °；The substrate is the ito glass that size is slightly larger than cover plate, ITO layer On be etched with electrode, the electrode is located in microchannel AE sections and is closely bonded therewith, including on be spaced each other one Electrode and two lower electrodes.

2. micro-fluidic chip according to claim 1, which is characterized in that described microchannel AB sections, AC sections and AD sections, AE The width ratio of section is 1: 2.0-3.0.

3. micro-fluidic chip according to claim 1 or 2, which is characterized in that described microchannel AB sections, AC sections of width Be 200 microns, AD sections, AE sections of width be 450 microns.

4. micro-fluidic chip according to claim 1, which is characterized in that ∠ BAC is 60 °.

5. micro-fluidic chip according to claim 1, which is characterized in that top electrode and the spacing of lower electrode are micro- for 30~60 Meter, the spacing between two lower electrodes is 15-30 microns.

6. the production method of the micro-fluidic chip of a kind of pair of cell multi-angle mechanical meaurement, which is characterized in that include the following steps:

S1, a top electrode being spaced each other and two lower electrodes are etched in the ITO layer of ito glass as substrate；

S2, it is lithographically derived anode membrane, reverse mould obtains the microchannel PDMS, and the microchannel AB sections of AD section for being located relatively at center line with AC sections is right Claim setting, AE section are AD section of straight line extension, AB sections and AC sections of width having the same, AD sections and AE sections of width having the same Spend and be wider than AB sections and AC sections, the end B, C-terminal, the end D and the end E of microchannel respectively correspond the first inlet of setting, the second inlet, Third inlet and liquid outlet, ∠ BAC are 30 ° -60 °；

S3, ito glass substrate is aligned with PDMS cover plate by alignment stage, so that electrode is located in microchannel AE sections, Close bonding is to get micro-fluidic chip.

7. manufacturing method according to claim 1, which is characterized in that described microchannel AB sections, AC sections and AD sections, AE sections Width ratio be 1: 2.0-3.0.

8. production method according to claim 6 or 7, which is characterized in that described microchannel AB sections, AC sections of width be 200 microns, AD sections, AE sections of width be 450 microns.

9. manufacturing method according to claim 1, which is characterized in that ZBAC is 60 °.

10. manufacturing method according to claim 1, which is characterized in that top electrode and the spacing of lower electrode are micro- for 30~60 Meter, the spacing between two lower electrodes is 15-30 microns.

Technical field

The present invention relates to micro-fluidic chip and its manufactures, and in particular to a kind of pair of cell multi-angle mechanical meaurement it is micro-fluidic Chip and preparation method thereof.

Background technique

Micro-fluidic (Microfluidics) technology is one of important branch of MEMS technology, is biology, chemistry, medicine The basic operation units such as sample preparation, reaction, separation, the detection of analytic process are integrated on the chip of one piece of micro-meter scale, from It is dynamic to complete analysis overall process.Due to its great potential in the fields such as biology, chemistry, medicine, have been developed as a biology, The brand-new research field of the subject crossings such as chemistry, medicine, fluid, electronics, material, machinery is that current development is more rapidly more One of subject crossing Environment Science technology, in life science, clinical medicine, chemical, pharmacy, food hygiene, environment measuring There is important application with the subjects such as monitoring, information science and signal detection.

The carrier that microflow control technique usually will use microanalysis device to realize as technology, and micro-fluidic chip is all kinds of micro- It analyzes with the fastest developing speed in device.Micro-fluidic chip is using MEMS technology, in silicon, quartz, glass or high molecular polymerization It processes various micro-structures, such as microchannel, micro reaction pool, microelectrode functional unit on object substrate, is then connected with microchannel Logical Micropump, micro-valve, micro- liquid storage device, micro- detecting element etc. are maximum with fluid conveying, control and the component for detecting function for monitoring Micro- complete point by process integration such as dilution, addition reagent, sampling, reaction, separation dispersion, detection, monitoring on chip of limit ground Analysis system.The area of micro-fluidic chip is usually a few to tens of square centimeters, and microchannel size is typically in micron order or close Grade.Micro-fluidic chip have it is at low cost, can batch micro operations, the advantages such as easy to operate, reproducible, reliability is high.

The biochip (micro-arrays) generally believed at present, such as genetic chip, protein-chip are miniflow The lattice array type hybridization hybrid chip that amount is zero, function is very limited, belongs to the specific type of micro-fluidic chip (micro-chip), micro- Fluidic chip has the function of wider type and purposes, can develop biocomputer, gene and protein sequencing, matter The analysis systems such as spectrum and chromatography, become the particularly important technical foundation of systems biology especially system genetics.Although in the presence of A variety of micro-fluidic chips and its application, but the micro-fluidic chip of multi-angle mechanical meaurement is not carried out to cell.

Summary of the invention

In view of this, the purpose of the present invention is to provide the micro-fluidic chip of a kind of pair of cell multi-angle mechanical meaurement, with Make up the deficiency of existing micro-fluidic chip type and its application.

Specifically, the micro-fluidic chip of the invention to cell multi-angle mechanical meaurement, is encapsulated by substrate and cover plate, The cover plate is the dimethyl silicone polymer (PDMS) equipped with microchannel, wherein during microchannel AB section and AC sections are located relatively at The AD section of heart line is symmetrical arranged, AE section be AD section straight line extension, AB sections and AC sections of width having the same, AD sections and AE sections Width having the same and it is wider than AB sections and AC sections, the end B, C-terminal, the end D and the end E of microchannel respectively correspond the first feed liquor of setting Mouth, the second inlet, third inlet and liquid outlet, ∠ BAC are 30 ° -60 °；The substrate is size slightly larger than cover plate ITO (tin indium oxide) glass, is etched with electrode on indium tin oxide layer, and the electrode is located in microchannel AE sections and therewith closely Bonding, including a top electrode being spaced each other and two lower electrodes.

In above-mentioned technical proposal, described microchannel AB sections, AC sections with AD sections, AE sections of width ratio is 1: 2.0-3.0.

Further, described microchannel AB sections, AC sections of width be 200 microns, AD section, AE sections of width it is micro- for 450 Rice.

In above-mentioned technical proposal, ∠ BAC is 60 °.

In above-mentioned technical proposal, the spacing of top electrode and lower electrode is 30~60 microns, the spacing between two lower electrodes It is 15-30 microns.

Present invention provides a kind of production methods of above-mentioned micro-fluidic chip to cell multi-angle mechanical meaurement, specifically Include the following steps:

(1) top electrode being spaced each other is etched on the indium tin oxide layer of ito glass and two lower electrodes are made For substrate；

(2) it is lithographically derived anode membrane, reverse mould obtains the microchannel PDMS, the microchannel AB sections of AD for being located relatively at center line with AC sections Section is symmetrical arranged, and AE sections are AD sections of straight line extension, and AB sections and AC sections of width having the same, AD sections with AE sections with identical Width and be wider than AB sections and AC sections, the end B, C-terminal, the end D and the end E of microchannel respectively correspond setting the first inlet, second into Liquid mouth, third inlet and liquid outlet, ∠ BAC is 30 ° -60 °, and (∠ BAC indicates to be formed between microchannel AB and microchannel AC Angle)；

The specific manufacture craft of the microchannel PDMS is as follows:

Photoresist is coated on silicon wafer；

The silicon wafer for having coated photoresist is toasted；

Silicon wafer after baking is subjected to photoetching process by preset mask plate and obtains the mold of fluid channel；

It impales the silicon wafer surrounding to form box-like cavity；

Resin solution is poured into the box-like cavity；

By it is described it is resin solution curing after, separated with silicon wafer, obtain the miniflow channel layer containing fluid channel.

3) ito glass substrate is aligned with PDMS cover plate by alignment stage, so that electrode is located at microchannel AE sections Interior, close bonding is to get micro-fluidic chip.

In above-mentioned technical proposal, described microchannel AB sections, AC sections with AD sections, AE sections of width ratio is 1: 2.0-3.0.

Further, described microchannel AB sections, AC sections of width be 200 microns, AD section, AE sections of width it is micro- for 450 Rice.

In above-mentioned technical proposal, ∠ BAC is 60 °.

In above-mentioned technical proposal, the spacing of top electrode and lower electrode is 30~60 microns, the spacing between two lower electrodes It is 15-30 microns.

Compared with prior art, the invention has the following beneficial effects:

Present invention firstly discloses a kind of micro-fluidic chips that multi-angle mechanical meaurement can be carried out to cell, pass through control When angle, capture of the realization electrode to cell stretch cell in different directions, can connect non-the width of microchannel Under conditions of touching to cell carry out multi-angle mechanical meaurement, the micro-fluidic chip is once purged reusable, have it is at low cost, The advantages that measurement is quick, small to cell damage.The production method of micro-fluidic chip of the invention is simple, high production efficiency, can be with It realizes batch production, there is wide commercial application prospect.

Detailed description of the invention

Fig. 1 is crude ito glass structural schematic diagram；

Fig. 2 is the ito glass schematic diagram of substrate structure etched after electrode；

Fig. 3 is the electrode partial enlarged view of Fig. 2 ito glass substrate；

Fig. 4 is the PDMS structural schematic diagram for making microchannel；

Fig. 5 is the schematic perspective view of the micro-fluidic chip of bonding packaging；

Fig. 6 is the top view of the micro-fluidic chip of bonding packaging；

Fig. 7 is the schematic diagram of internal structure of the micro-fluidic chip of bonding packaging；

Wherein, 1 it is ito glass substrate, 2 be electrode, 21 be top electrode, 22/23 be lower electrode, 3 be cover plate, 4 is micro- logical Road, 51 be the first inlet, 52 be the second inlet, 53 be third inlet, 6 be liquid outlet.

Specific embodiment

Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out detailed retouch It states.