CN203639470U

CN203639470U - Microfluidic chip for researching biological behaviors of cell migration after cell injury

Info

Publication number: CN203639470U
Application number: CN201320881780.6U
Authority: CN
Inventors: 聂富强; 吴圆丽
Original assignee: Suzhou Institute of Nano Tech and Nano Bionics of CAS
Current assignee: Suzhou Institute of Nano Tech and Nano Bionics of CAS
Priority date: 2013-12-30
Filing date: 2013-12-30
Publication date: 2014-06-11
Anticipated expiration: 2023-12-30

Abstract

The utility model discloses a microfluidic chip. The microfluidic chip comprises a base material and a channel formed in the base material, wherein the channel comprises a main culture passage, at least two sample feed ports are formed in one end of the main culture passage, and the other end of the main culture passage is communicated with a sample discharge port. The microfluidic chip can realize cell implantation and wound preparation by using a laminar flow phenomenon between microfluids under the control of gravity drive produced by the liquid level difference between the sample feed ports and the sample discharge port, so that biological behaviors of cell migration after cell injury can be intuitively observed under a microscope, and a foundation is laid for cell biology for researching cell wound healing. The microfluidic chip can be used for simultaneously realizing noninvasive wound preparation of the cells and facilitating uninterrupted observation of the biological behaviors of the cell migration in real time, and can be applied to various cell migration researches.

Description

A kind of study cell and be wound after the micro-fluidic chip of cell migration biological behaviour

Technical field

The application relate to a kind of study cell and be wound after the micro-fluidic chip of cell migration biological behaviour.

Background technology

Cell migration is a dynamic complex process that instructed by a large amount of chemistry and physical signalling, is playing the part of important role in numerous physiology and pathologic process.Cell migration needs the cooperation of internal and external factor, the signaling molecule outside external factor phalangeal cell, and internal factor is the signal transducting system of phalangeal cell and cytoskeleton and the molecular motor of execution motion, participates in addition the various molecules that adhesion plaque forms.In the research of cell migration biological behaviour, traditional method has obtained remarkable achievement as conventional cell migration models such as cell wound healing, induction diffusion process, Boyden cell/Transwell cells in the research of cell migration biological behaviour, but, because it is difficult to break through truly in analogue body the problems such as microenvironment (as wall shear stress) and concentration gradient be unstable, make cell migration process leave over lower a lot of unsolved mystery.Because of the limitation of self, conventional art cannot meet complicated vital process and the numerous needed high order of magnitude experiment number of tera incognita and huge data analysis, makes cell migration biological study present bottleneck.

There is the cell migration of wound class, also referred to as cell wound healing class cell migration campaign, cultivate monolayer cell and also produce wound area, then study the be wound phenomenon of place's peripheral cell migration of this wound.The method is the simplest, and the basic cell biology method of cheapness and high duplication is also one of method of directly studying in vitro the earliest the characteristics such as cell migration speed, persistence and polarization simultaneously.In existing macroscopical technology, in order to obtain cell wound, conventionally adopt physics cut mode, its shortcoming is: the wound obtaining is crude, and edge of wound cell can damage, and affects cytoactive, residual cell fragment is many, is not easy to dynamic observational study, and circulation ratio is not high.

Since the nineties in 20th century, an important trend of natural science and engineering development is to stride forward to microminiaturization.Micro-fluidic chip technology is by propositions such as Switzerland scientist Manz, and this technology is to drive/control the analysis test platform as core technology, modern analysis detection technique as means taking the micro-basis of micro-processing, microfluid.Micro-fluidic chip technology is different from traditional macroscopical technical concept completely, it has dwindled the volume installing, improve analysis efficiency, reduced reagent consumption, avoid environmental pollution, as medical science, chemistry, life science, environment etc. are applied widely, demonstrate huge development potentiality and using value in numerous scientific domains with microminiaturized, integrated, high-throughput, high-precision feature.In recent years, because micro-fluidic chip technology has grid type two dimension or three-dimensional channel structure and micron-sized channel size, can simultaneously on time and space, control the features such as fluid and advantage, utilize micro-fluidic chip technology to study cell migration biological behaviour to be subject to many investigators' concern.

Utility model content

The purpose of this utility model provides a kind of micro-fluidic chip, overcomes existing physics cut mode and produces the shortcoming that cell wound is crude, have fringing effect.

For achieving the above object, the utility model provides following technical scheme:

A kind of study cell and be wound after the micro-fluidic chip of cell migration biological behaviour, described micro-fluidic chip comprises base material and is formed at the raceway groove in described base material, described raceway groove comprises cultivates main channel, and one end of this cultivation main channel is provided with at least two injection ports, and the other end is communicated with an outlet.

Preferably, after above-mentioned research cell is wound, in the micro-fluidic chip of cell migration biological behaviour, extend in horizontal plane described cultivation main channel, and described injection port vertically extends.

Preferably, after above-mentioned research cell is wound, in the micro-fluidic chip of cell migration biological behaviour, described outlet vertically extends.

Preferably, after above-mentioned research cell is wound, in the micro-fluidic chip of cell migration biological behaviour, between described each injection port and cultivation main channel, be also communicated with a buffer channel, described buffer channel and described cultivation main channel are located in the same horizontal plane.

Preferably, after above-mentioned research cell is wound, in the micro-fluidic chip of cell migration biological behaviour, all described buffer channels have identical bore.

Preferably, after above-mentioned research cell is wound in the micro-fluidic chip of cell migration biological behaviour, described base material comprises the first base material and the second base material that stacked on top of one another arranges, described main channel and buffer channel are opened in the upper surface of described the second base material, and described injection port and outlet run through the upper and lower surface of described the first base material or the second base material.

Preferably, after above-mentioned research cell is wound, in the micro-fluidic chip of cell migration biological behaviour, one end of described cultivation main channel is provided with three injection ports.

Compared with prior art, the utility model has the advantage of: the utility model proposes a kind of be wound based on micro-fluidic chip research cell after the method for cell migration biological behaviour, be convenient to observe, equipment is simple, Direct Sampling, sample and reagent dosage little, sample is little without transfer, sample crossed contamination probability, can study more realistically cell migration biological behaviour, be with a wide range of applications in association areas such as cytobiology, genetics and drug screenings.

Brief description of the drawings

In order to be illustrated more clearly in the embodiment of the present application or technical scheme of the prior art, to the accompanying drawing of required use in embodiment or description of the Prior Art be briefly described below, apparently, the accompanying drawing the following describes is only some embodiment that record in the application, for those of ordinary skill in the art, do not paying under the prerequisite of creative work, can also obtain according to these accompanying drawings other accompanying drawing.

Figure 1 shows that the structural representation of micro-fluidic chip in the utility model specific embodiment;

Figure 2 shows that the sectional view along dotted line in Fig. 1;

Figure 3 shows that in the utility model specific embodiment, cultivating main channel forms the schematic diagram after cell monolayer;

Figure 4 shows that the schematic diagram of the digested formation wound of cell monolayer in the utility model specific embodiment;

Figure 5 shows that the structural representation of micro-fluidic chip in the utility model the second embodiment.

Embodiment

Below in conjunction with the accompanying drawing in the utility model embodiment, the technical scheme in the utility model embodiment is described in detail, obviously, described embodiment is only the utility model part embodiment, instead of whole embodiment.Based on the embodiment in the utility model, the every other embodiment that those of ordinary skill in the art obtain under the prerequisite of not making creative work, belongs to the scope that the utility model is protected.

Figure 1 shows that the structural representation of micro-fluidic chip in the utility model embodiment.Figure 2 shows that along the sectional view of dotted line in Fig. 1.

Shown in ginseng Fig. 1 and Fig. 2, micro-fluidic chip comprises base material 1 and is formed at the raceway groove 2 in base material 1.

Base material 1 comprises the first base material 11 and the second base material 12 that stacked on top of one another arranges, the material of base material 1 is selected from PMMA(polymethylmethacrylate), PC(polycarbonate), COC resin, ABS(acrylonitrile-butadiene-styrene copolymer), glass, quartz or copper, be preferably PMMA, the material of the first base material 11 and the second base material 12 can be identical, also can be different.

Raceway groove 2, for being formed at the groove on the second base material 12 upper surfaces, comprising and cultivates main channel 21 and be communicated in three buffer channels 22 cultivating 21 one end, main channel.Cultivate main channel 21 and three buffer channels 22 and be positioned at same plane.The cross section of cultivating main channel 21 and buffer channel 22 is preferably rectangle, and the length of three buffer channels 22 and bore are all identical.In addition, the bore of cultivating main channel 21 is preferably three times of single buffer channel 22 bores, like this, and when solution enters cultivation main channel 21 from three buffer channels 22, can not become suddenly large or diminish and change original kinestate because of bore, the interface of solution remains straight line simultaneously.

On the first base material 11, run through and be provided with three injection ports 111 and an outlet 112, three injection ports 111 are communicated in respectively the entrance of three buffer channels 22 and vertically extend, and outlet 112 is communicated in to be cultivated the outlet of main channel 21 and vertically extends.

In other embodiments, base material can be also only one flat plate, and injection port and outlet are arranged at dull and stereotyped side; Buffer channel 22 can only be provided with two or be greater than three, and corresponding opening for feed is also set to two or be greater than three.

The making method of above-mentioned micro-fluidic chip is as follows:

(1) use computer aided design software (CAD) to design microchannel and the microstructure of micro-fluidic chip;

(2) cultivate main channel, buffer channel, injection port and outlet by laser ablation, pressure sintering, moulding method, the micro-processing of numerical control or soft lithographic technique in the preparation of PMMA substrate surface;

(3) will be prepared with the micro-fluidic chip base material of cultivating main channel and buffer channel and cut into the size (making the second base material) of 4x4cm, again the base material that is prepared with injection port and outlet is cut into the size (making the first base material) of 4x4cm, finished base material is cleaned with tap water, distilled water respectively, and by the residual spot such as fingerprint, oil stain of ethanol substrate surface, naturally dry;

(4) by Heat Sealing technology or by double-deck adhesive membrane viscosity glue, the first base material and the second base material are alignd under the microscope, carry out involution, make the micro-fluidic chip for studying cell migration biological behaviour.

Cell migration biological behaviour after utilizing above-mentioned micro-fluidic chip research cell to be wound, method is as follows:

The rifle head of four applicable circular cone bores is inserted to the import and export of chip, add cell solution from three injection ports, control the poor height of liquid between three injection ports and outlet, make the cell solution cell cultures main channel of slowly flowing through, cell is in slowly by cultivation main channel process, absorption rests on the inwall (on the surrounding inwall of the diapire of passage or passage) of the cell cultures passage of involution, when absorption cell quantity account for cultivate main channel 1/10th time, complete the implantation of cell, micro-fluidic chip is put into cell culture incubator and cultivate.

With microscopic examination, form (shown in ginseng Fig. 3) after cell monolayer when cultivating main channel, add respectively containing trypsin 10% from left side injection port, middle injection port and right side injection port), PBS damping fluid, trypsin 10%) cell culture fluid, control the poor height of liquid of three injection ports and outlet, thereby key-course Flow Velocity, make trypsinase Rapid Flow through cultivating the cell monolayer of main channel, the cell being flow through by trypsin solution is therefore digested, flow out microchannel with fluid, complete standby without formulating (shown in ginseng Fig. 4) of cell wound.Can control in addition left and right sides injection port trypsin 10%) form the intermediate cell layer wound of different in width with the poor height of liquid of middle injection port PBS damping fluid, be convenient to study the impact of cell-cell interaction on wound.Finally slowly rinse residual trypsinase with PBS solution, after cell wound forms and stablizes, microscope can be in real time incessantly observation of cell be wound after the biological behaviour of wound cell migration, the cell of cell wound is all active, the cell migration phenomenon after can real observation biomass cells wound forming.

In aforesaid method, also can in the injection port of the left and right sides, pass into PBS damping fluid, middle injection port passes into trypsin 10% simultaneously).Thereby trypsinase object is to make intercellular proteolysis that cell is dismissed, and it can be substituted by EDTA or collagenase equally.PBS damping fluid can use the cell balance salts solution that Hanks liquid, D-Hanks liquid, Dulbecco etc. are conventional to replace.

Shown in ginseng Fig. 5, in the utility model the second embodiment, cultivate main channel, buffer channel, injection port and outlet and be all formed on the first base material, the second base material is a blank flat board.

In sum, the utility model is by controlling under injection port and outlet liquid level pressure reduction generation segregation drive, form layers flow phenomenon between the liquid of different feeds mouth, utilize the laminar flow phenomenon between microfluid, trypsinase to the digestion of cell adhesion layer under, realize forming without wound of the implantation of cell and cell wound, the biological behaviour of cell migration after can studying intuitively cell under the microscope and being wound, for research wound healing and cell migration biology lay the foundation.

It should be noted that, in this article, relational terms such as the first and second grades is only used for an entity or operation to separate with another entity or operational zone, and not necessarily requires or imply and between these entities or operation, have the relation of any this reality or sequentially.And, term " comprises ", " comprising " or its any other variant are intended to contain comprising of nonexcludability, thereby the process, method, article or the equipment that make to comprise a series of key elements not only comprise those key elements, but also comprise other key elements of clearly not listing, or be also included as the intrinsic key element of this process, method, article or equipment.The in the situation that of more restrictions not, the key element being limited by statement " comprising ... ", and be not precluded within process, method, article or the equipment that comprises described key element and also have other identical element.

The above is only the application's embodiment; it should be pointed out that for those skilled in the art, do not departing under the prerequisite of the application's principle; can also make some improvements and modifications, these improvements and modifications also should be considered as the application's protection domain.

Claims

One kind study cell and be wound after the micro-fluidic chip of cell migration biological behaviour, it is characterized in that: described micro-fluidic chip comprises base material and is formed at the raceway groove in described base material, described raceway groove comprises cultivates main channel, one end of this cultivation main channel is provided with at least two injection ports, and the other end is communicated with an outlet.
Research cell according to claim 1 be wound after the micro-fluidic chip of cell migration biological behaviour, it is characterized in that: extend in horizontal plane described cultivation main channel, and described injection port vertically extends.
Research cell according to claim 1 and 2 be wound after the micro-fluidic chip of cell migration biological behaviour, it is characterized in that: described outlet vertically extends.
Research cell according to claim 1 and 2 be wound after the micro-fluidic chip of cell migration biological behaviour, it is characterized in that: between described each injection port and cultivation main channel, be also communicated with a buffer channel, described buffer channel and described cultivation main channel are located in the same horizontal plane.
Research cell according to claim 4 be wound after the micro-fluidic chip of cell migration biological behaviour, it is characterized in that: all described buffer channels have identical bore.
Research cell according to claim 4 be wound after the micro-fluidic chip of cell migration biological behaviour, it is characterized in that: described base material comprises the first base material and the second base material that stacked on top of one another arranges, described cultivation main channel and buffer channel are opened in the upper surface of described the second base material, and described injection port and outlet run through the upper and lower surface of described the first base material or the second base material.
Research cell according to claim 1 be wound after the micro-fluidic chip of cell migration biological behaviour, it is characterized in that: one end of described cultivation main channel is provided with three injection ports.