CN105606535B - Surface plasmon resonance sensing chip and cellular response detection system and method - Google Patents
Surface plasmon resonance sensing chip and cellular response detection system and method Download PDFInfo
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- G—PHYSICS
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/55—Specular reflectivity
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- G01N21/553—Attenuated total reflection and using surface plasmons
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Abstract
The invention discloses a kind of surface plasmon resonance sensing chips, comprising: culture chamber, the culture chamber have culture liquid flow inlet and culture fluid outlet；Heating device, the heating device includes the sandwich with two layers of heat-conducting metal and the high resistivity film being clipped in the middle and has communicating structure, wherein the culture chamber is embedded in the communicating structure；Culture solution flow channel, the culture solution flow channel make the culture solution in it flow through a surface of the heating device before the culture liquid flow inlet for entering the culture chamber；And the prism positioned at the culture chamber lower part and with the close sealing-in of the culture chamber, noble metal film is coated on the surface that the prism is engaged with the culture chamber.The sensing chip can also include transparent thermally insulating housing, and its culture chamber can be layered.Thereby, it is possible to maintain the temperature of cell growing environment for a long time and uniformly and stably, and then promote the accuracy of cellular response detection.
Description
Technical field
The present invention relates to field of biological detection more particularly to one kind can for a long time, stablize, maintain evenly cell growth
The surface plasmon resonance sensing chip of required temperature.
Background technique
Cell is the basic unit of life entity.Cell detection facilitates the deep essence for understanding vital movement process, favorably
In medical diagnosis on disease and drug development.Surface plasma body resonant vibration (SPR) sensing technology has real-time, sensitive, excellent without label etc.
Point has become grinding nearly ten years using the response to different chemotherapy regimens of SPR detection cellular response, especially cancer cell
Study carefully hot spot.
Fig. 1 is the schematic diagram of the SPR sensorgram chip system 10 of the prior art.For convenience of explanation, optics is eliminated in figure
Optical coupling device and angular adjustment device in part and A/D conversion circuit and computer in data acquisition process part
Deng, and only carry out the representative as optical system and data acquisition processing system using light source 120 and optical detection unit 130.
Sensing chip 110 includes lid glass, arises from inflow entrance and the finally fluid channel of outflux and positioned at lower part for exciting
The prism of SPR, wherein the upper surface of prism is coated with noble metal film, usually golden film.Biomolecule or cell inoculation can be existed
The surface of metal film will lead to metal when these biomolecule or cell change due to interacting with culture solution
The change of membrane superficial tissue, so as to cause the change at the angle SPR.According to the change value at the angle SPR, it will be able to obtain corresponding biological detection
Result.In order to preferably carry out biological culture, need to control culture environment.As shown, near fluid channel
It is disposed with the flow sensor for sensing flow, for the temperature sensor of sensing temperature and for tieing up fluid temperature (F.T.)
It holds to the heater of preset temperature (usually 37 degrees Celsius).
Temperature needed for maintaining cell normal growth for a long time is one of the essential condition for realizing SPR cell detection.Existing
Have in technology, it will usually (no using multi-disc heater (the TEC cooling piece usually produced in batches) package cell culture chamber surrounding
It is only 4 shown in Fig. 1), by comparing the difference of actual temperature and preset temperature inside sensing chip, control TEC refrigeration
The current direction and size of piece are realized and are controlled the temperature of sensing chip.
Since SPR sensorgram chip needs to place prism in bottom, and need top giving over to watch window, therefore cause
Sensing chip top and the bottom temperature control measures are insufficient, and the exterior temperature change of sensing chip will lead to the fluctuation of its internal temperature, influence
The stability detected for a long time.In addition, temperature when due to culture solution sample introduction is (usually for the sensing chip based on flowing culture
For room temperature) it is low compared with preset temperature (usually 37 degrees Celsius), the flowing of culture solution, which will lead to, generates temperature ladder inside sensing chip
Degree.Different degrees of influence can be caused to the cell growth of different location in sensing chip in this way, and then influence testing result
Consistency.Finally, the TEC device used now belongs to batch production device, shape is usually single, is unfavorable for structure of controlling temperature and sets
Meter.
Thus, it is desirable to which a kind of can overcome the surface plasma body resonant vibration of above-mentioned at least one disadvantage in the prior art to pass
Sense chip.
Summary of the invention
A technical problem to be solved by this invention is to overcome the above-mentioned deficiency of the prior art, provides a kind of new use
In the temprature control method and sensing chip of SPR cell detection, can for a long time, stablize, maintain evenly needed for cell growth
Temperature, realize response of the long-time SPR detection cell to environmental stimuli.
According to an aspect of the invention, there is provided a kind of surface plasmon resonance sensing chip, comprising: culture chamber,
Culture chamber has culture liquid flow inlet and culture fluid outlet；Heating device, heating device include have two layers heat-conducting metal and
The sandwich for the high resistivity film being clipped in the middle and have communicating structure, wherein culture chamber is embedded in communicating structure；Training
Nutrient solution flow channel, culture solution flow channel flow through the culture solution in it before the culture liquid flow inlet for entering culture chamber
One surface of heating device；And the prism positioned at culture chamber lower part and with the close sealing-in of culture chamber, prism connect with culture chamber
Noble metal film is coated on the surface of conjunction.
Preferably, sensing chip can also include transparent or semitransparent thermally insulating housing.
Preferably, culture solution flow channel is located in thermally insulating housing, and heating device heats dress in thermally insulating housing
The surface set and culture solution flow channel are closely adjacent.
Preferably, sensing chip can also include: the culture solution flow pass in thermally insulating housing, culture solution outflow
Channel is connected to the culture fluid outlet of culture chamber.
Preferably, culture chamber includes the perfusate chamber on top and the culturing room of lower part, and is clipped in the perforated membrane among two Room,
Wherein culture liquid flow inlet and culture fluid outlet are located at perfusate chamber.
Preferably, the inner cavity of perfusate chamber can be contour cavity, and the horizontal cross-section of contour cavity is by a triangle, one
Square and a semicircle are spliced, and culture liquid flow inlet is located at the vertex of triangle, and culture fluid outlet is located at half
Circular apex.Correspondingly, culturing room can be cylindrical body and be located at the lower part of square.
Preferably, culturing room has cell inlet and cell outlet, and cell inlet and cell outlet are being cultivated
Liquid stream when perfusate chamber through closing.
Preferably, perfusate chamber, culturing room and perforated membrane can be by being molded one-pass molding.
According to another aspect of the present invention, a kind of cellular response detection system is provided, comprising: surface as described above
Plasma resonance sensing chip；Optical module, for emitting and modulating for injecting surface plasmon resonance sensing chip
In prism incident light；And data acquisition and processing (DAP) component, for receive the data of the incident light from optical module with
And the reflected light from prism data and the data of acquisition are handled to obtain the testing result of cellular response.
According to another aspect of the present invention, a kind of cellular response detection method is provided, comprising: inject cell as above
The culture chamber of the surface plasmon resonance sensing chip, wherein cell is grown on the noble metal film of prism upper surface；
Culture solution is set to flow continuously through the perfusate chamber of sensing chip, wherein culturing room carries out culture solution exchange by interlayer；Keep incident light logical
Prism incidence is crossed to the upper surface of prism；And it acquires incident and reflected light data and is pocessed to obtain the inspection of cellular response
Survey result.
In this way, having the heating device of sandwich by introducing, heating device is enabled more easily to adapt to train
Support the various shape of chamber.In addition, since culture solution flows through a surface of heating device before flowing into culture chamber, it can
The temperature difference of culture solution and culture chamber is eliminated, guarantees that the temperature in culture chamber is stablized.
In addition, the present invention also passes through the culture chamber for introducing thermally insulating housing and " two Room " structure, further isolated cell growth
The heat of internal environment and external overall situation conducts.
Detailed description of the invention
Disclosure illustrative embodiments are described in more detail in conjunction with the accompanying drawings, the disclosure above-mentioned and its
Its purpose, feature and advantage will be apparent, wherein in disclosure illustrative embodiments, identical reference label
Typically represent same parts.
Fig. 1 is the schematic diagram of the SPR sensorgram chip system of the prior art.
Fig. 2 is the sectional view of SPR sensorgram chip according to an embodiment of the invention.
Fig. 3 is the sectional view of the heating device according to an embodiment of the invention with sandwich.
Fig. 4 is the top view of sandwich shown in Fig. 3.
Fig. 5 is the sectional view of the culture chamber with " two Room " structure according to an embodiment of the present invention.
Fig. 6 is the top view of the culture chamber with " two Room " structure according to an embodiment of the present invention.
Specific embodiment
The preferred embodiment of the disclosure is more fully described below with reference to accompanying drawings.Although showing the disclosure in attached drawing
Preferred embodiment, however, it is to be appreciated that may be realized in various forms the disclosure without the embodiment party that should be illustrated here
Formula is limited.On the contrary, these embodiments are provided so that this disclosure will be more thorough and complete, and can be by the disclosure
Range is completely communicated to those skilled in the art.
Since existing SPR sensorgram chip can not be that SPR cellular response detects good temperature control accurately to simulate carefully
The environment of intracellular growth, the invention proposes a kind of SPR sensorgram chips with the novel structure for capableing of good temperature control.Different from existing
There is the TEC cooling piece that semiconductor refrigerating principle is utilized in technology, the present invention is carried out by the heating device with sandwich
Temperature control.In addition, it is default to be preheated to firstly flow through the surface of heating device by making culture solution before entering culture chamber
Temperature, the temperature gradient caused by eliminating due to culture solution flowing inside culture solution perfusate chamber.In addition, using transparent or semitransparent
Thermally insulating housing wrap up sensing chip in addition to prism, further enhance sensing core while guaranteeing cell growth observability
The ability of piece resistance exterior temperature change.Finally, having the culture chamber of " two Room " structure by introducing, it is further reduced fluid temperature
Influence of the degree variation to the cell growth being located in lower part cell culture room.
Fig. 2 is the sectional view of SPR sensorgram chip 210 according to an embodiment of the invention.The sensing chip 210 can wrap
Include thermally insulating housing 201, heating device 202, culture chamber 204, culture solution flow channel 205 and culture solution flow pass 206.
Culture chamber 201 has culture liquid flow inlet and cultivates outflow and discharge that fluid outlet is used for culture solution.As schemed
Show, culture liquid flow inlet and culture fluid outlet can regard the tail end of culture solution flow channel 205 as respectively and cultivate liquid stream
The head end in channel 206 out.
Heating device 202 includes sandwich and tool with two layers of heat-conducting metal and the high resistivity film being clipped in the middle
There is communicating structure.Culture chamber 210 is embedded in the communicating structure of heating device.Fig. 3 is tool according to an embodiment of the invention
There is the sectional view of the heating device 302 of sandwich.
In the example of fig. 3, heating device 302 is by being located at the intermediate high resistivity of upper and lower two layers of heat-conducting metal and load
Film composition.In one example, this double layer of metal can be the copper sheet that there are hole (or any other shapes) in the identical centre of size,
Size is identical and the intermediate PVDF thin film for equally having hole (or any other shapes) for can be of being clipped in the middle.It is injected by control
The size of current of PVDF thin film, the heating power of control heating ring.Hole among sandwich (or meets any of culture chamber
Other shapes) communicating structure is formed, culture chamber (for example, culture chamber 210) can be embedded in the communicating structure.Fig. 4 is
The top view of sandwich shown in Fig. 3.
As shown in figure 4, the outer shape of back-shaped heating device is a square, inside have a communicating structure, use
In placement culture chamber.It should be understood that sandwich can be added according to the needs of culture chamber shape and manufacturing process
Work is at arbitrary shape.For example, the heating device can be the interior cuboid knot for having circular through hole if culture chamber is cylindrical
Structure, it might even be possible in a ring.
Sensing chip 210 shown in Fig. 2 is returned to, culture solution flow channel 205 connects with the culture liquid flow inlet of culture chamber,
Its at least part is located on the upper surface of heating device, so that the culture solution in it flows through before entering the culture chamber
The surface, is thus preheated to preset temperature.Culture solution flow pass 206 connects with the culture fluid outlet of culture chamber, is used for
The discharge of culture solution.
Preferably, sensor chip can also include thermally insulating housing 201.Thermally insulating housing 201 wraps up entire sensing chip and removes
Other parts outside prism, for being thermally isolated with external environment.Thermally insulating housing 201 can be it is transparent or translucent, by
This guarantees the observability inside culture chamber.Culture solution flow channel 205 can be located at 201 in thermally insulating housing, heating device 202
Then can embedded in thermally insulating housing 201 in advance there are groove in, and the upper surface of heating device can be with culture solution flow channel
205 is close adjacent, to ensure that the heat of heating device is sufficiently conducted to the culture solution flowed through.Culture solution flow pass 206 can also
To be located at 201 in thermally insulating housing, the effect of isolation environment temperature is thus further realized.
Prism 204 can be right-angle prism or Dove prism.The prism 204 is located at 203 lower part of culture chamber, and close with it
Sealing-in is to form a closed chamber.At least part of 204 upper surface of prism is coated with noble metal film, which constitutes training simultaneously
Support the lower surface of chamber 203.For example, the noble metal film can be the silverskin or golden film of one layer of 20-50nm thickness, thereon for biology
The growth of molecule/cell.For example, targeted cancerous cells can be made in golden film in the test for the chemotherapy effect of cancer cell
Upper growth, and cancer is inferred according to the variation of the incident light and reflected light that pass through prism (and the variation at the corresponding angle SPR)
Thus the upgrowth situation of cell gives accurate assessment to chemotherapy effect.
Preferably, culture chamber 203 can have the internal structure of layering and be grown with further isolation environment temperature to cell
Influence.The culture chamber with preferred internal structure will be described in conjunction with Fig. 5 and Fig. 6 as follows.
Fig. 5 is the sectional view of the culture chamber 503 according to an embodiment of the invention with " two Room " structure.As schemed
Show, " two Room " structure is by the culture solution perfusate chamber 511 on top and the cell culture chamber 513 of lower part, and is clipped among two Room
Interlayer 512 (for example, porous membrane) composition.Liquid environment needed for culturing room 513 is used to provide cell growth (therefore can also claim
For cell growth chamber), perfusate chamber 511 is trained for the heat transfer at the top of isolated cell culturing room with external environment, while with cell
It supports room and carries out mass exchange.Total is nested in the central hollow section of heating ring, and four side walls and heating ring are in close contact,
Heat transfer for isolated cell culturing room side and external environment.
Perfusate chamber 511 has culture liquid flow inlet 515 and culture fluid outlet 516, for flowing in and out for culture solution.
Although entrance is located at the top of perfusate chamber in figure, it will be appreciated by those skilled in the art that can also be based on actually answering
With any position that entrance 515 and 516 is located to top perfusate chamber 511, as long as its inflow that can be realized culture solution and stream
Out.Entrance 515 and 516 preferably respectively with culture solution flow channel and flow pass (for example, the 205 of Fig. 2 and 206)
It is connected, to realize the inflow and discharge of culture solution.In other embodiments, one of flow channel and flow pass also can be omitted
Or both, entrance is directly open on the shell, but the temperature control effect of this structure is not so good as the structure with fluid channel.
Perforated membrane 512 between perfusate chamber 511 and culturing room 513, ensure while avoiding two Room direct-connected two Room that
This is in fluid communication.In this way, culture solution, after the perfusate chamber 511 for entering top, a part is diffused into lower part through perforated membrane 512
In culturing room 513, the waste that nutrition and exchange discharge cell metabolism for supplementing cell consumption generate, remaining is directly from outlet
516 discharges.Intracorporal nutrition exchange velocity can be simulated by adjusting the number, size and distribution in hole on perforated membrane, from
And help to build and be similar to intracorporal cell growing environment, thus more accurate testing result is provided.In other embodiments
In, interlayer 512 can have other structures, such as can be film with seam, not the mass exchange film of physically opening or other
Spacer structure, as long as its exchange that can be realized the suitable speed of culture solution in culturing room 513.
Culturing room 513 is tightly engaged into prism 504.Although being shown in figure and side seal under culture chamber for clear signal
A part of the prism 504 connect, it should be appreciated that, although prism 504 is the necessary part of SPR sensorgram chip, be not
A part of culture chamber 503.
Preferably, culturing room 513 can have cell inlet 517 and cell outlet 518.Above-mentioned 517 He of entrance
518 injection and discharge (that is, each cell culture is before and after) for cell, and closed when cultivating cell, that is,
The closing when culture solution flows through perfusate chamber of cell inlet 517 and cell outlet 518.Entrance 517 and 518 is located at culturing room
On 513 side walls, expression the two mouthfuls in Fig. 5 shown in dotted line are arranged on the direction vertical with paper, that is, 517 He of entrance
518 line is vertical with the line of entrance 515 and 515.Although avoiding arranging the opening of perfusate chamber 511 and culturing room 513
Be conducive to the processing of sensing chip in the same direction, it will be understood by those skilled in the art that discrepancy can be arranged arbitrarily
The position of mouth 517 and 518, as long as it can facilitate the injection and discharge of cell.In another embodiment, may be used not
Cell inlet 517 and cell outlet 518 are set, but filled using culture liquid flow inlet 515 and culture fluid outlet 516
Infuse culture solution before, the injection of cell is realized by the opening 515 or 516 of perfusion compartment, and after cell culture terminates
Use the discharge (for example, conduit by protruding into) of 515 or 516 realization cell of opening.
Perfusate chamber 511, interlayer 512 and culturing room 513 can be manufactured to any suitable size.Although Fig. 5 is shown
Cross-sectional width is greater than the perfusate chamber 511 of culturing room 513, it will be appreciated by those skilled in the art that can to fill
Flow chamber and culturing room 513 have same cross-sectional width, for example, the upper and lower part of the cylindrical body separated by interlayer.
The shape of perfusate chamber and culturing room can be reasonably selected, to advanced optimize two Room to internal cell growing environment
Simulation.Fig. 6 is the top view of the culture chamber 603 according to an embodiment of the invention with " two Room " structure.Implement at one
In example, Fig. 5 can regard the sectional view splitted along the line segment AA ' in Fig. 6 as, that is, culture chamber 603 can be with culture chamber
503 indicate identical construction.But it is understood that culture chamber 603 and 602 can also be different, for example, can be by the structure of Fig. 6
It makes and regards a preferred embodiment based on culture chamber 503 as.
As shown, the outside of perfusate chamber 611 can be a cuboid, and its internal cavities can be the high pitched tunes such as one
Body.The horizontal cross-section of the contour cavity is spliced by a triangle, a square and a semicircle, and culture solution flows into
Mouth 615 is located at the vertex of the triangle, and culture fluid outlet 616 is located at the semicircular apex.Preferably, described
Triangle can be the isosceles triangle that apex angle is 135 degree.
Using the particular configuration, culture solution can be made after inflow entrance 615 flows into perfusate chamber 611, firstly flow through three
Angular cavity forms uniform laminar flow；Rectangular enclosure is again passed through, a part is diffused into down through interlayer 612 (for example, porous membrane)
In the culturing room 613 in portion, the waste of nutrition and exchange discharge cell metabolism generation for supplementing cell consumption；Remaining culture solution
Semicircular cavities (arc-shaped construction can disturb to avoid any angle bring flow field) is flowed to, is finally discharged from outflux 616.
Culturing room 613 is located at the lower part of the square, for example, the inner cavity of culturing room 613 can be diameter and it is above-mentioned just
The long equal cylindrical body of Quad Sides.Interlayer 612 is located among culture solution perfusate chamber 611 and cell culture chamber 613.Interlayer 612 can
To be perforated membrane, the round micropore for being d comprising n diameter, the substance for culture solution perfusate chamber 611 and cell culture chamber 613
Exchange.
As a result, by the way that culture chamber to be divided into " two Room ", influence of the external environment to cell culture can further be isolated.Separately
Outside, by the way that the shape and size of " two Room " and interlayer are rationally arranged, the growing environment of cell in vivo can be more accurately simulated,
To further improve the accuracy of testing result.
Fig. 2-6 has been referred to above and has given the specific descriptions of invention and preferred embodiments thereof, will have been provided as follows according to this
The specific Production Example of the surface plasmon resonance sensing chip of inventive principle.
Thermally insulating housing (for example, 201 of Fig. 2) can be made of transparent plastics such as PMMA or PS, and interior includes a culture solution
Perfusion access road and one outlet channel.The length of thermally insulating housing can be 56mm, 60mm, 12mm respectively；And it cultivates
The circle that it is 1mm for the rectangular or diameter of 1mm X 1mm that the section of liquid exit and entrance, which can be,.
Heating device is (for example, the heating " back-shaped ring " 302) of 202, Fig. 3 of Fig. 2 and 4 is by two identical " back-shaped
Ring " copper sheet 311 and 313, and the PVDF thin film 312 being clipped in the middle form.The outer dimension of copper sheet and pvdf membrane is 27mm X
27mm X 1mm (long X wide X high), the rectangle of central hollow section 12mm X 9mm.The heating ring is nested in thermally insulating housing, is added
Hot ring upper surface and culture solution flow path are closely adjacent, for preheating to the culture solution that will flow into cell culture chamber.
Cell culture chamber (for example, 603 of 503 and Fig. 6 of 203, Fig. 5 of Fig. 2) is primary by injection molding by PDMS polymer
Molding.The cell culture chamber is by a culture solution perfusate chamber and a cell growth chamber, and is clipped in the perforated membrane among two Room
Composition.The external dimensions of culture solution perfusate chamber is 12mm X 9mm X 2mm (long X wide X high), and internal cavities are the circle of diameter 4mm
Cylinder, it is preferable that internal cavities can also be as shown in FIG. 6 and be spelled by a triangle, a square and one are semicircle
Connect composition: vertex angle is 135 °, opposite side side length 4mm；Square side length is 4mm；Semicircle diameter is 4mm.Culturing room
The external dimensions of (that is, cell growth chamber) is 12mm X 9mm X 1mm (long X wide X high), and internal cavities are the cylinder of diameter 4mm
Body.The height and diameter ratio (aspect ratio) of cell growth chamber are 1:4, and in other examples, aspect ratio can be in 3:1 to 5:1
Between.Porous membrane is with a thickness of 0.1mm, the circular through hole for being 0.2mm containing 9 diameters.
Prism can be right-angle prism or Dove prism；Material is BK7 or ZF5 glass；Upper surface is coated with the gold of 20-50nm
Film.Prism upper surface and the close sealing-in of cell growth chamber form airtight chamber, and cell, which is attached in golden film, to be grown.
The SPR sensorgram chip and its preferred embodiment of invention as described above.Sensing chip according to the present invention can
For a long time, temperature needed for stablizing, maintaining evenly cell growth realizes long-time SPR detection cell to the sound of environmental stimuli
It answers.
Using sensing chip of the invention, a kind of more accurate effective biological detection system can be obtained, for example, cell
Respond detection system.The system may include above-mentioned sensing chip and its preferred embodiment of the invention, can also include optics
Component and data acquisition and processing (DAP) component.Optical module can emit and modulate the incident light for injecting the prism, and
Data acquisition and processing (DAP) component then can receive the incident light data from optical module and the reflected light data from prism
And the data of acquisition are handled.By being subject to analysis processing to the data of acquisition, more accurate biology inspection can be obtained
It surveys as a result, for example, the testing result of cellular response.
Likewise it is possible to realize a kind of novel cell using above-mentioned surface plasmon resonance sensing chip of the invention
Detection method.When carrying out cell detection, can first by cell injection sensing chip culturing room (it is, for example, possible to use Fig. 6
Cell inlet 617 and cell outlet 618 carry out cell injection and discharge), cell be seeded in culturing room and
It is grown on the noble metal film of prism upper surface.In the incubation of cell, culture solution is made to flow continuously through the perfusion of sensing chip
Room, and the culturing room for being located at lower layer then carries out culture solution exchange by interlayer.When needing to detect cellular response, light beam can be made
Entrance prism, and the incident light according to caused by the variation the cell adhered state on noble metal film and reflected light relationship
Change (variation at the angle SPR), to obtain the corresponding testing result of cellular response.In addition, can be discharged intracavitary at the end of culture
Liquid is simultaneously cleaned, so that next time uses.
Using sensing chip and cell detection method of the invention can for a long time, stablize, to maintain evenly cell raw
Long required temperature, and then realize response of the long-time SPR detection cell to environmental stimuli, especially cancer cell is to different chemotherapy
The response of scheme obtains the drug corresponding information of cell.Biomedical and pharmaceuticals researcher can carry out accordingly medical diagnosis on disease,
Drug discovery and exploitation, modality of cancer treatment optimization etc..
Various embodiments of the present invention are described above, above description is exemplary, and non-exclusive, and
It is not limited to disclosed each embodiment.Without departing from the scope and spirit of illustrated each embodiment, for this skill
Many modifications and changes are obvious for the those of ordinary skill in art field.The selection of term used herein, purport
In the principle, practical application or improvement to the technology in market for best explaining each embodiment, or make the art
Other those of ordinary skill can understand each embodiment disclosed herein.
Claims (8)
1. a kind of surface plasmon resonance sensing chip, comprising:
Culture chamber, the culture chamber has culture liquid flow inlet and culture fluid outlet, wherein the culture chamber includes top
The culturing room of perfusate chamber and lower part, and it is clipped in the perforated membrane among two Room, wherein the culture solution inflow entrance and the culture
Fluid outlet is located at the perfusate chamber；
Heating device, the heating device include the sandwich with two layers of heat-conducting metal and the high resistivity film being clipped in the middle
And there is communicating structure, wherein the culture chamber is embedded in the communicating structure；
Culture solution flow channel, the culture solution flow channel make the culture solution in it in the training for entering the culture chamber
A surface of the heating device is flowed through before nutrient solution inflow entrance；And
Prism positioned at the culture chamber lower part and with the close sealing-in of the culture chamber, the prism are engaged with the culture chamber
Noble metal film is coated on surface,
Wherein, the inner cavity of the perfusate chamber is contour cavity, the horizontal cross-section of the contour cavity by a triangle, one just
A rectangular and semicircle is spliced, and the culture liquid flow inlet is located at the vertex of the triangle, the culture liquid stream
Outlet is located at the semicircular apex, and the culturing room is cylindrical body and the lower part for being located at the square.
2. sensing chip as described in claim 1, wherein the sensing chip further includes transparent or semitransparent thermally insulating housing.
3. sensing chip as claimed in claim 2, wherein the culture solution flow channel is located in the thermally insulating housing, it is described
Heating device is in the thermally insulating housing, and the surface of the heating device and the culture solution flow channel are close
It is adjacent.
4. sensing chip as claimed in claim 3, further includes:
Culture solution flow pass in the thermally insulating housing, the culture liquid stream of the culture solution flow pass and the culture chamber
Outlet.
5. sensing chip as described in claim 1, wherein the culturing room has cell inlet and cell outlet, it is described
Cell inlet and cell outlet closing when culture solution flows through the perfusate chamber.
6. sensing chip as described in claim 1, wherein the perfusate chamber, the culturing room and the perforated membrane pass through note
Mould one-pass molding.
7. a kind of cellular response detection system, comprising:
Such as surface plasmon resonance sensing chip of any of claims 1-6；
Optical module, the optical module emit and modulate for injecting the institute in the surface plasmon resonance sensing chip
State the incident light of prism；And
Data acquisition and processing (DAP) component, the data acquisition and processing (DAP) component receive the number of the incident light from the optical module
Accordingly and the data of the reflected light from the prism and the data of acquisition are handled to obtain the detection knot of cellular response
Fruit.
8. a kind of cellular response detection method, comprising:
Cell is injected into the culture chamber such as surface plasmon resonance sensing chip of any of claims 1-6,
Described in cell grown on the noble metal film of the prism upper surface；
Culture solution is set to flow continuously through the perfusate chamber of the sensing chip, wherein the culturing room is by being clipped in the perfusate chamber and institute
It states the perforated membrane among culturing room and carries out culture solution exchange；
Incident light is set to pass through the prism incidence to the upper surface of the prism；And
Acquisition incidence and reflected light data are simultaneously pocessed to obtain the testing result of cellular response.
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