CN105606535A - Surface plasma resonance sensor chip and cell response detection system and method - Google Patents
Surface plasma resonance sensor chip and cell response detection system and method Download PDFInfo
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- CN105606535A CN105606535A CN201610094869.6A CN201610094869A CN105606535A CN 105606535 A CN105606535 A CN 105606535A CN 201610094869 A CN201610094869 A CN 201610094869A CN 105606535 A CN105606535 A CN 105606535A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
- G01N21/552—Attenuated total reflection
- G01N21/553—Attenuated total reflection and using surface plasmons
Abstract
The invention discloses a surface plasma resonance sensor chip comprising a culture cavity, a heating device, a culture solution inflow channel and a prism. The culture cavity is provided with a culture solution inflow port and a culture solution outflow port. The heating device comprises two layers of heat conductive metal and a high-resistivity membrane interlayer structure clamped between the two layers of heat conductive metal and is of a through structure, wherein the culture cavity is embedded in the through structure. The culture solution inflow channel makes a culture solution in the culture solution inflow channel flow through one surface of the heating device before entering the culture solution inflow port of the culture cavity. The prism is located on the lower portion of the culture cavity and closely connected with the culture cavity in a sealing mode, the surface, connected with the culture cavity, of the prism is plated with a precious metal membrane. The sensor chip also comprises a transparent heat-insulating shell, and the culture cavity can be layered. Thus, the temperature of a cell growing environment can be evenly and stably maintained for a long time, and cell response detection accuracy is improved.
Description
Technical field
The present invention relates to field of biological detection, relating in particular to one can be for a long time, stable, equablyMaintain the temperature required surface plasmon resonance sensing chip of Growth of Cells.
Background technology
Cell is the elementary cell of life entity. Cell detection contributes to the deep vital movement process understoodEssence, is conducive to medical diagnosis on disease and drug development. Surface plasma body resonant vibration (SPR) sensing technologyHave real-time, sensitive, without advantages such as marks, utilize SPR detect cellular response, particularly cancer is thinThe response of born of the same parents to different chemotherapy regimens, has become study hotspot nearly ten years.
Fig. 1 is the schematic diagram of the SPR sensing chip system 10 of prior art. For convenience of description,In figure, save optical coupling device and angular adjustment device and the data acquisition process in opticatorA/D change-over circuit and computer etc. in part, and only with light source 120 and optical detection unit 130Carry out the representative as optical system and data acquisition processing system respectively. Sensing chip 110 comprises lidGlass, arise from inflow entrance and finally flow export fluid passage and be positioned at bottom for exciting SPR'sPrism, wherein the upper surface of prism is coated with noble metal film, normally golden film. Can be by biomolecule orCell is seeded in the surface of metal film, when these biomolecule or cell are owing to interacting with nutrient solutionAnd while changing, can cause the change of metallic film surface structure, thereby cause the change at SPR angle.According to the change value at SPR angle, just can draw the result of corresponding biological detection. In order better to carry outBiological culture, need to control culture environment. As shown in the figure, near fluid passage, be furnished withFor the flow sensor of sensed flow, for the temperature sensor of sensing temperature and for flowingTemperature is maintained until the heater of preset temperature (being generally 37 degrees Celsius).
Maintaining for a long time the required temperature of cell normal growth, is to realize important of SPR cell detectionOne of part. In the prior art, conventionally can use multi-disc heater (the normally TEC of batch productionCooling piece) parcel cell culture chamber surrounding (being not only 4 shown in Fig. 1), by relatively passingThe difference of the inner actual temperature of sense chip and preset temperature, controls the sense of current of TEC cooling piece with largeLittle, realize the temperature control to sensing chip.
Because SPR sensing chip need to be placed prism in bottom, and top need to be given over to observation windowMouthful, therefore cause sensing chip top and the bottom temperature control measures deficiency, the exterior temperature change meeting of sensing chipCause the fluctuation of its internal temperature, the long-term stability detecting of impact. In addition, for training based on flowingFoster sensing chip, the temperature (being generally room temperature) during due to nutrient solution sample introduction is (logical compared with preset temperatureBe often 37 degrees Celsius) low, the mobile meeting of nutrient solution causes the inner thermograde that produces of sensing chip. ThisSample can cause impact in various degree to the Growth of Cells of diverse location in sensing chip, and then impact inspectionSurvey the uniformity of result. Finally, the TEC device using now belongs to batch production device, and profile is logicalChang Danyi, is unfavorable for structure of controlling temperature design.
Thus, need a kind of surface that can overcome above-mentioned at least one shortcoming of the prior art etc. fromDaughter resonance sensing chip.
Summary of the invention
A technical problem to be solved by this invention is to overcome the above-mentioned deficiency of prior art, providesThe new temperature-controlled process for SPR cell detection and a sensing chip, can be for a long time, surelyDetermine, maintain the required temperature of Growth of Cells equably, realize long-time SPR detection cell and sting to external worldSharp response.
According to an aspect of the present invention, provide a kind of surface plasmon resonance sensing chip, bagDraw together: culture chamber, culture chamber has nutrient solution inflow entrance and nutrient solution flow export; Heater, heatingDevice comprises the sandwich of the high resistivity film that has two-layer heat-conducting metal and be clipped in the middle and hasCommunicating structure, wherein culture chamber is embedded in communicating structure; Nutrient solution flow channel, nutrient solution flows intoPassage makes nutrient solution in it heater of flowing through before entering the nutrient solution inflow entrance of culture chamberA surface; And be positioned at culture chamber bottom the prism with the tight sealing-in of culture chamber, prism and cultivationOn the surface that chamber engages, be coated with noble metal film.
Preferably, sensing chip can also comprise transparent or semitransparent thermally insulating housing.
Preferably, nutrient solution flow channel is positioned at thermally insulating housing, and heater is embedded in thermally insulating housing,And the surface of heater and nutrient solution flow channel are closely adjacent.
Preferably, sensing chip can also comprise: be positioned at the nutrient solution flow pass of thermally insulating housing,This nutrient solution flow pass is communicated with the nutrient solution flow export of culture chamber.
Preferably, culture chamber comprises the perfusate chamber on top and the culturing room of bottom, and is clipped in two RoomBetween perforated membrane, wherein nutrient solution inflow entrance and nutrient solution flow export are positioned at perfusate chamber place.
Preferably, the inner chamber of perfusate chamber can be contour cavity, and the horizontal cross-section of contour cavity is by oneTriangle, a square and a semicircle are spliced, and nutrient solution inflow entrance is positioned at leg-of-muttonDrift angle place, nutrient solution flow export is positioned at semicircular summit place. Correspondingly, culturing room can be cylinderBody and be positioned at foursquare bottom.
Preferably, culturing room has cell inlet and cell outlet, cell inlet and cell rowOutlet is sealed in the time that nutrient solution is flowed through perfusate chamber.
Preferably, perfusate chamber, culturing room and perforated membrane can be by injection moulding one-shot formings.
According to another aspect of the present invention, provide a kind of cellular response detection system, having comprised: asThe surface plasmon resonance sensing chip of the above; Optical module, is used for penetrating for launching and modulatingEnter the incident light of the prism in surface plasmon resonance sensing chip; And data acquisition and processing (DAP) groupPart, for receiving from the data of the incident light of optical module and from the catoptrical data of prismAnd the data that gather are processed to obtain the testing result of cellular response.
According to another aspect of the present invention, provide a kind of cellular response detection method, having comprised: willCell injects the culture chamber of surface plasmon resonance sensing chip as above, and wherein cell is at ribOn the noble metal film of mirror upper surface, grow; Make nutrient solution Continuous-flow cross the perfusate chamber of sensing chip, whereinCulturing room carries out nutrient solution exchange by interlayer; Make incident light be incident to the upper surface of prism by prism;And gather incident and reflected light data and processed to obtain the testing result of cellular response.
Like this, there is the heater of sandwich by introducing, heater can more be heldChange places and adapt to the various shapes of culture chamber. In addition, because nutrient solution was flowed through and added before flowing into culture chamberThe surface that hot charging is put, therefore can eliminate the temperature difference of nutrient solution and culture chamber, ensures culture chamberInterior temperature stabilization.
In addition, the present invention is also by introducing the culture chamber of thermally insulating housing and " two Room " structure, furtherThe heat conduction of isolated cell growth internal environment and outside overall situation.
Brief description of the drawings
In conjunction with the drawings disclosure illustrative embodiments is described in more detail, of the present disclosureAbove-mentioned and other object, Characteristics and advantages will become more obvious, wherein, exemplary in the disclosureIn embodiment, identical reference number represents same parts conventionally.
Fig. 1 is the schematic diagram of the SPR sensing chip system of prior art.
Fig. 2 is the sectional view of SPR sensing chip according to an embodiment of the invention.
Fig. 3 is the sectional view according to an embodiment of the invention with the heater of sandwich.
Fig. 4 is the top view of sandwich shown in Fig. 3.
Fig. 5 is the sectional view according to one embodiment of the invention with the culture chamber of " two Room " structure.
Fig. 6 is the top view according to one embodiment of the invention with the culture chamber of " two Room " structure.
Detailed description of the invention
Preferred embodiment of the present disclosure is described below with reference to accompanying drawings in more detail. Although aobvious in accompanying drawingShow preferred embodiment of the present disclosure, but should be appreciated that, can realize the disclosure with various formsAnd the embodiment that should do not set forth here limits. On the contrary, providing these embodiments is in order to makeThe disclosure is more thorough and complete, and the scope of the present disclosure intactly can be conveyed to this areaTechnical staff.
Because existing SPR sensing chip cannot detect good temperature control for SPR cellular responseThe accurately environment of analog cell growth, the present invention proposes a kind of well novelty knot of temperature control that hasThe SPR sensing chip of structure. Be different from the TEC cooling piece that utilizes semiconductor refrigerating principle in prior art,The present invention carries out temperature control by the heater with sandwich. In addition, by making trainingNutrient solution, before entering culture chamber, first flows through the surface of heater to be preheated to preset temperature, eliminatesDue to the thermograde of the mobile nutrient solution perfusate chamber inside causing of nutrient solution. In addition, use transparent orTranslucent thermally insulating housing wraps up the sensing chip except prism, is ensureing Growth of Cells observabilityFurther strengthen the ability of sensing chip opposing exterior temperature change simultaneously. Finally, have by introducingThe culture chamber of " two Room " structure, further reduces fluid temperature variations to being positioned at bottom cell culture chamberThe impact of interior Growth of Cells.
Fig. 2 is the sectional view of SPR sensing chip 210 according to an embodiment of the invention. This sensingChip 210 can comprise that thermally insulating housing 201, heater 202, culture chamber 204, nutrient solution flow intoPassage 205 and nutrient solution flow pass 206.
Culture chamber 201 has nutrient solution inflow entrance and nutrient solution flow export outflow and the row for nutrient solutionGo out. As shown in the figure, nutrient solution inflow entrance and nutrient solution flow export can regard respectively that nutrient solution flows into asThe head end of the tail end of passage 205 and nutrient solution flow pass 206.
Heater 202 comprises the interlayer of the high resistivity film that has two-layer heat-conducting metal and be clipped in the middleStructure and there is communicating structure. Culture chamber 210 is embedded in the communicating structure of heater. Fig. 3It is the sectional view according to an embodiment of the invention with the heater 302 of sandwich.
In the example of Fig. 3, heater 302 is by being arranged in upper and lower two-layer heat-conducting metal and loadingBetween high resistivity film composition. In an example, this double layer of metal can be measure-alike centreThe copper sheet of porose (or arbitrarily other shapes), what be clipped in the middle can be measure-alike and middle sameThe PVDF film of porose (or arbitrarily other shapes). Inject the electric current of PVDF film by controlSize, the heating power of control heating ring. Hole in the middle of sandwich (or meets appointing of culture chamberOther shapes of anticipating) form a communicating structure, culture chamber (for example, culture chamber 210) just can embedIn this communicating structure. Fig. 4 is the top view of sandwich shown in Fig. 3.
As shown in Figure 4, the external shape of back-shaped heater is a square, and its inside has oneCommunicating structure, for placing culture chamber. It should be understood that sandwich can be according to culture chamber shapeThe needs of shape and manufacturing process and be processed to arbitrary shape. For example, if culture chamber is cylindrical,This heater has the rectangular structure of manhole in can being, even can be ringwise.
Get back to the sensing chip 210 shown in Fig. 2, the cultivation of nutrient solution flow channel 205 and culture chamberLiquid inflow entrance joins, and its at least a portion is positioned on the upper surface of heater, to make the training in itNutrient solution this surface of flowing through before entering described culture chamber, is preheated to preset temperature thus. Nutrient solutionFlow pass 206 joins with the nutrient solution flow export of culture chamber, for the discharge of nutrient solution.
Preferably, sensor chip can also comprise thermally insulating housing 201. Thermally insulating housing 201 wraps up wholeIndividual sensing chip other parts except prism, for isolating with the heat of external environment. Thermally insulating housing 201Can be transparent or translucent, ensure thus the observability of culture chamber inside. Nutrient solution flows intoPassage 205 can be positioned at thermally insulating housing 201, and 202 of heaters can be embedded in thermally insulating housing 201In the groove leaving in advance, and the upper surface of heater can be tight with nutrient solution flow channel 205Close adjacent, fully conduct to the heat of guaranteeing heater the nutrient solution flowing through. Nutrient solution flows out logicalRoad 206 also can be positioned at thermally insulating housing 201, further realizes thus the effect of isolation environment temperature.
Prism 204 can be right-angle prism or Dove prism. This prism 204 is positioned at culture chamber 203 timesPortion, and sealing-in tight with it is to form a sealing chamber. At least a portion of prism 204 upper surfacesBe coated with noble metal film, this part forms the lower surface of culture chamber 203 simultaneously. For example, this noble metal filmCan be silverskin or the golden film that one deck 20-50nm is thick, on it for the growth of biomolecule/cell.For example, in the test of the chemotherapy effect for cancer cell, can make targeted cancerous cells on golden filmGrowth, and according to through the incident light of prism and catoptrical variation (and corresponding SPR angleChange) infer and thus chemotherapy effect is given the upgrowth situation of cancer cell to assess accurately.
Preferably, the internal structure that culture chamber 203 can have layering is with further isolation environment temperatureThe impact of cell growth. As follows in connection with Fig. 5 and Fig. 6 to thering is the culture chamber of preferred internal structureDescribed.
Fig. 5 be have according to an embodiment of the invention " two Room " structure culture chamber 503 cutFace figure. As shown in the figure, " two Room " structure is by the nutrient solution perfusate chamber 511 on top and the cell of bottomCulturing room 513, and be clipped in interlayer 512 (for example, the porous membrane) composition in the middle of two Room. CultivateChamber 513 is for the required liquid environment of Growth of Cells (therefore also can be described as cell growth chamber) is provided,Perfusate chamber 511, for the heat conduction of isolated cell culturing room top and external environment, is trained with cell simultaneouslyMass exchange is carried out in foster chamber. Total is nested in the middle hollow parts of heating ring, four sidewalls withHeating ring close contact, for the heat conduction of isolated cell culturing room side and external environment.
Perfusate chamber 511 has nutrient solution inflow entrance 515 and nutrient solution flow export 516, for nutrient solutionInflow and outflow. Although in figure, gateway is located to perfusate chamber top, those skilled in the art canTo understand, also can gateway 515 and 516 be located to top perfusate chamber 511 based on practical applicationAny position, as long as its can realize nutrient solution inflow and flow out. Gateway 515 and 516Preferably respectively with nutrient solution flow channel and flow pass (for example, 205 of Fig. 2 and 206) phaseConnect, to realize inflow and the discharge of nutrient solution. In other embodiment, also can omit flow channelWith one of flow pass or both, with by direct gateway opening on shell, but the temperature of this structureControl effect is not as having the structure of fluid passage.
Perforated membrane 512, between perfusate chamber 511 and culturing room 513, is being avoided direct-connected same in two RoomTime guarantee two Room fluid communication with each other. Like this, nutrient solution is entering after the perfusate chamber 511 on top, and onePart sees through perforated membrane 512 and is diffused in the culturing room 513 of bottom, for supplementing the battalion of cell consumptionSupport and exchange and discharge the refuse that cellular metabolism produces, all the other are directly from exporting 516 discharges. Can pass throughAdjust number, size and the distribution in hole on perforated membrane, the nutrition exchange velocity in can analogue body, fromAnd contribute to build the Growth of Cells environment being similar in body, testing result more is accurately provided thus.In other embodiments, interlayer 512 can have other structures, for example, can be film with seam, not haveThe mass exchange film of physically opening, or other spacer structure, as long as it can realize culturing room 513The exchange of the suitable speed of middle nutrient solution.
Culturing room 513 closely engages with prism 504. Although for clear signal, there is shown withA part for the prism 504 of culture chamber downside sealing-in, should be appreciated that, although prism 504 is SPRThe necessary part of sensing chip, but be not a part for culture chamber 503.
Preferably, culturing room 513 can have cell inlet 517 and cell outlet 518. OnState gateway 517 and 518 injections for cell and discharge (before, each cell is cultivated and is started andAfter finishing), and seal in the time of cultured cell, that is, and cell inlet 517 and cell outlet 518In the time that flowing through perfusate chamber, seals nutrient solution. Gateway 517 and 518 is positioned on culturing room's 513 sidewalls,These two mouths of the expression being shown in dotted line in Fig. 5 are arranged in the direction vertical with paper, that is, and and gateway517 is vertical with 515 line with gateway 515 with 518 line. Although avoid perfusate chamber 511In same direction, be conducive to the processing of sensing chip with the aperture arrangement of culturing room 513, but abilityField technique personnel should be appreciated that, can lay out arbitrarily the position of entrance 517 and 518, as long as its energyEnough facilitate the injection of cell and discharge. In another embodiment, cell note also can be setEntrance 517 and cell outlet 518, but using nutrient solution inflow entrance 515 and nutrient solution to flow outBefore mouth 516 perfusion cultures liquid, realize the injection of cell by the opening 515 or 516 of perfusion compartment,And cell cultivate finish after use opening 515 or 516 realize cell discharge (for example, lead toCross the conduit stretching into).
Perfusate chamber 511, interlayer 512 and culturing room 513 can be manufactured to any suitable size.Although showing cross-sectional width, Fig. 5 is greater than the perfusate chamber 511 of culturing room 513, this area skillArt personnel are understandable that, can make perfusate chamber and culturing room 513 have same cross-sectional width,For example, the cylindrical upper and lower being separated by interlayer.
Can choose reasonable perfusate chamber and the shape of culturing room, further to optimize two Room to cells in vivoThe simulation of growing environment. Fig. 6 is the training according to an embodiment of the invention with " two Room " structureSupport the top view in chamber 603. In one embodiment, Fig. 5 can regard as along the line segment in Fig. 6AA ' cuts the sectional view obtaining open, that is, culture chamber 603 can represent identical structure by culture chamber 503.But be understandable that, culture chamber 603 and 602 also can be different, for example, can be by the structure of Fig. 6Make a preferred embodiment of regarding as based on culture chamber 503.
As shown in the figure, the outside of perfusate chamber 611 can be a cuboid, and its internal cavities canIt is a contour cavity. The horizontal cross-section of these high cavitys is by a triangle, square andIndividual semicircle is spliced, and nutrient solution inflow entrance 615 is positioned at described leg-of-mutton drift angle place, nutrient solutionFlow export 616 is positioned at described semicircular summit place. Preferably, described triangle can be that drift angle isThe isosceles triangle of 135 degree.
Use this particular configuration, can make nutrient solution in inflow entrance 615 flows into perfusate chamber 611After, first flow through triangular cavity, form even laminar flow; Flow through again rectangular enclosure, a part see through everyLayer 612 (for example, porous membrane) are diffused in the culturing room 613 of bottom, disappear for supplementing cellThe refuse that cellular metabolism produces is discharged in the nutrition of consumption and exchange; All the other nutrient solutions flow to semicircular cavities (circleThe flow field disturbance that arc structure can avoid any angle to bring), finally discharge from flow export 616.
Culturing room 613 is positioned at described foursquare bottom, and for example, the inner chamber of culturing room 613 can beThe cylinder that diameter equates with the above-mentioned square length of side. Interlayer 612 is positioned at nutrient solution perfusate chamber 611 HesIn the middle of cell culture chamber 613. Interlayer 612 can be perforated membrane, comprises the circle that n diameter is dMicropore, for the mass exchange of nutrient solution perfusate chamber 611 and cell culture chamber 613.
Thus, by culture chamber being divided into " two Room ", can further isolate external environment to cellThe impact of cultivating. In addition, by the shape and size of reasonable setting " two Room " and interlayer, can be more smartTrue analog cell growing environment in vivo, thus the accuracy of testing result improved further.
Below provided the specific descriptions of invention and preferred embodiments thereof with reference to figure 2-6, as follows willProvide the concrete Production Example of surface plasmon resonance sensing chip in accordance with the principles of the present invention.
Thermally insulating housing (for example, Fig. 2 201) can be made up of the transparent plastic such as PMMA or PS,In it, comprise a nutrient solution perfusion access road and an exit passageway. The length and width of thermally insulating housing,Height can be respectively 56mm, 60mm, 12mm; And the cross section of nutrient solution exit and entrance can beSquare for 1mmX1mm, or the diameter circle that is 1mm.
Heater (for example, 202 of Fig. 2, Fig. 3 and 4 heating " back-shaped ring " 302) is by twoIndividual identical " back-shaped ring " copper sheet 311 and 313, and the PVDF film 312 being clipped in the middleComposition. The appearance and size of copper sheet and pvdf membrane is 27mmX27mmX1mm (the wide X of long XHigh), the rectangle of middle hollow parts 12mmX9mm. This heating ring is nested in thermally insulating housing,Heating ring upper surface and nutrient solution stream are closely adjacent, for the cultivation to by flowing into cell culture chamberLiquid carries out preheating.
Cell culture chamber (for example, 203 of Fig. 2,503 and Fig. 6 of Fig. 5 603) by PDMSPolymer is by injection moulding one-shot forming. This cell culture chamber is by a nutrient solution perfusate chamber and a cellGrowth room, and be clipped in the perforated membrane composition in the middle of two Room. The external dimensions of nutrient solution perfusate chamber is 12MmX9mmX2mm (the wide X of long X is high), internal cavities is the cylinder of diameter 4mm,Preferably, internal cavities can also be as shown in Figure 6 by a triangle, square andIndividual semicircle splicing composition: triangle drift angle is 135 °, opposite side length of side 4mm; The square length of side is4mm; Semicircle diameter is 4mm. The external dimensions of culturing room's (, cell growth chamber) is 12mmX9mmX1mm (the wide X of long X is high), internal cavities is the cylinder of diameter 4mm. CellThe height of growth room and diameter ratio (aspect ratio) are 1:4, and in other examples, aspect ratio can beBetween 3:1 to 5:1. Porous membrane thickness is 0.1mm, contains the circle that 9 diameters are 0.2mmThrough hole.
Prism can be right-angle prism or Dove prism; Material is BK7 or ZF5 glass; Upper surface platingThere is the golden film of 20-50nm. The tight sealing-in of prism upper surface and cell growth chamber forms airtight chamber, thinBorn of the same parents are attached on golden film and grow.
SPR sensing chip of the present invention described above and preferred embodiment thereof. According to biography of the present inventionSense chip can be for a long time, stable, maintain the required temperature of Growth of Cells equably, realize long-timeSPR detects the response that cell stimulates to external world.
Use sensing chip of the present invention, can obtain one biological detection system more accurately and effectively,For example, cellular response detection system. This system can comprise above-mentioned sensing chip of the present invention and excellentSelect embodiment, can also comprise optical module and data acquisition and processing (DAP) assembly. Optical module canLaunch and modulate the incident light for injecting described prism, data acquisition and processing (DAP) assembly can connectReceive from the incident light data of optical module and from the reflected light data of prism and to the data that gatherProcess. By in addition analyzing and processing of the data of obtaining, can obtain biological inspection more accuratelySurvey result, for example, the testing result of cellular response.
Similarly, can use above-mentioned surface plasmon resonance sensing chip of the present invention to realize a kind ofNovel cell detection method. In the time carrying out cell detection, can first cell be injected to sensing chipCulturing room (for example, can use the cell inlet 617 of Fig. 6 and cell outlet 618 to carry outThe injection of cell and discharge), cell is seeded in culturing room and at the noble metal film of prism upper surfaceUpper growth. In the incubation of cell, make nutrient solution Continuous-flow cross the perfusate chamber of sensing chip, andThe culturing room that is positioned at lower floor carries out nutrient solution exchange by interlayer. In the time that needs detect cellular response,Can make light beam entrance prism, and lead according to the variation of the cell attachment state due on noble metal filmThe variation (variation at SPR angle) of the incident light causing and reverberation relation, obtains the phase of cellular responseAnswer testing result. In addition, in the time cultivating end, can discharge intracavity liquid and be cleaned, forUsed next time.
Use sensing chip of the present invention and cell detection method just can be for a long time, stable, equablyMaintain the required temperature of Growth of Cells, and then realize long-time SPR and detect the sound that cell stimulates to external worldShould, the particularly response of cancer cell to different chemotherapy regimens, the medicine corresponding information of acquisition cell. RawThing medical science and pharmaceuticals researcher can carry out medical diagnosis on disease, drug discovery and exploitation accordingly, cancer is controlledTreat scheme optimization etc.
Below described various embodiments of the present invention, above-mentioned explanation is exemplary, not limitProperty, and be also not limited to disclosed each embodiment. At the model that does not depart from illustrated each embodimentEnclose in the situation with spirit many modifications and changes for those skilled in the artAll apparent. The selection of term used herein, is intended to explain best that each embodiment's is formerReason, practical application or the improvement to the technology in market, or make other common skill of the artArt personnel can understand the each embodiment disclosing herein.
Claims (10)
1. a surface plasmon resonance sensing chip, comprising:
Culture chamber, described culture chamber has nutrient solution inflow entrance and nutrient solution flow export;
Heater, described heater comprises the high resistance that has two-layer heat-conducting metal and be clipped in the middleThe sandwich of rate film and have communicating structure, wherein said culture chamber is embedded in described communicating structureIn;
Nutrient solution flow channel, described nutrient solution flow channel makes its interior nutrient solution described in enteringFlow through before the described nutrient solution inflow entrance of a culture chamber surface of described heater; And
Be positioned at described culture chamber bottom the prism with the tight sealing-in of described culture chamber, described prism and instituteState culture chamber engage surface on be coated with noble metal film.
2. sensing chip as claimed in claim 1, wherein said sensing chip also comprises transparent or halfTransparent thermally insulating housing.
3. sensing chip as claimed in claim 2, described in wherein said nutrient solution flow channel is positioned atIn thermally insulating housing, described heater is embedded in described thermally insulating housing, and the institute of described heaterState surface closely adjacent with described nutrient solution flow channel.
4. sensing chip as claimed in claim 3, also comprises:
Be positioned at the nutrient solution flow pass of described thermally insulating housing, this nutrient solution flow pass and described trainingThe nutrient solution flow export of supporting chamber is communicated with.
5. the sensing chip as described in any one in claim 1 to 4, wherein said culture chamber comprisesThe perfusate chamber on top and the culturing room of bottom, and be clipped in the perforated membrane in the middle of two Room, wherein said trainingNutrient solution inflow entrance and described nutrient solution flow export are positioned at described perfusate chamber place.
6. sensing chip as claimed in claim 5, the inner chamber of wherein said perfusate chamber is contour cavity,The horizontal cross-section of described contour cavity by a triangle, square and semicircle splicing andBecome, described nutrient solution inflow entrance is positioned at described leg-of-mutton drift angle place, and described nutrient solution flow export is positioned atDescribed semicircular summit place, described culturing room is cylinder and is positioned at described foursquare bottom.
7. sensing chip as claimed in claim 5, wherein said culturing room have cell inlet andCell outlet, described cell inlet and described cell outlet are at the nutrient solution described perfusate chamber of flowing throughIn time, seals.
8. sensing chip as claimed in claim 5, wherein said perfusate chamber, described culturing room andDescribed perforated membrane is by injection moulding one-shot forming.
9. a cellular response detection system, comprising:
Surface plasmon resonance sensing chip as described in any one in claim 1-8;
Optical module, described optical module is launched and is modulated and is used for injecting described surface plasma body resonant vibrationThe incident light of the described prism in sensing chip; And
Data acquisition and processing (DAP) assembly, described data acquisition and processing (DAP) assembly receives from described optics groupThe data of the incident light of part and from the catoptrical data of described prism and to gather data carry outProcess to obtain the testing result of cellular response.
10. a cellular response detection method, comprising:
Cell is injected to the surface plasma resonance sensing core as described in claim 1-8 any oneThe culture chamber of sheet, wherein said cell is grown on the described noble metal film of described prism upper surface;
Make nutrient solution Continuous-flow cross the perfusate chamber of described sensing chip, described in wherein said culturing room passes throughInterlayer carries out nutrient solution exchange;
Make incident light be incident to the upper surface of described prism by described prism; And
Gather incident and reflected light data and also processed to obtain the testing result of cellular response.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109297935A (en) * | 2018-11-30 | 2019-02-01 | 深圳大学 | A kind of surface plasma resonance image-forming device and method |
| CN109985677A (en) * | 2019-04-24 | 2019-07-09 | 浙江警察学院 | Modularization sample single channel detection chip component suitable for hand-held SPR detector |
| CN113764257A (en) * | 2021-08-25 | 2021-12-07 | 清华大学深圳国际研究生院 | Desorption electric spray ionization device based on 3D printing |
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| CN109297935B (en) * | 2018-11-30 | 2021-04-20 | 深圳大学 | Surface plasma resonance imaging device and method |
| CN109985677A (en) * | 2019-04-24 | 2019-07-09 | 浙江警察学院 | Modularization sample single channel detection chip component suitable for hand-held SPR detector |
| CN113764257A (en) * | 2021-08-25 | 2021-12-07 | 清华大学深圳国际研究生院 | Desorption electric spray ionization device based on 3D printing |
| CN113764257B (en) * | 2021-08-25 | 2024-01-12 | 清华大学深圳国际研究生院 | Desorption electrospray ionization device based on 3D printing |
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