CN209508217U - A kind of digital pcr fluorescence detection device - Google Patents
A kind of digital pcr fluorescence detection device Download PDFInfo
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- CN209508217U CN209508217U CN201822122338.0U CN201822122338U CN209508217U CN 209508217 U CN209508217 U CN 209508217U CN 201822122338 U CN201822122338 U CN 201822122338U CN 209508217 U CN209508217 U CN 209508217U
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- digital pcr
- optical filter
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Abstract
The utility model provides a kind of digital pcr fluorescence detection device, comprising: support base；Bracket；Objective table and optical module, the objective table include electronic slide unit, the fixed reaction chamber of the sample being fixed on electronic slide unit and the temperature-controlling module for regulating and controlling the fixed reaction chamber temperature of sample, and the fixed reaction chamber of sample is for accommodating PCR sample to be measured；Electronic slide unit can be along X, the moving in two dimensional directions of Y-axis；Optical module is arranged on bracket, including light source portion, optical filter wheel, object lens and ccd image sensor；The light beam that light source portion issues is irradiated in PCR sample to be measured after successively passing through optical filter wheel and object lens, after object lens and optical filter wheel, the fluorogenic quantitative detection to PCR sample to be measured is successively realized into ccd image sensor for the transmitting light that PCR sample to be measured is issued through irradiation.The device further includes digital pcr chip；The device can be realized the augmentation detection in situ and multiplex PCR detection of PCR.
Description
Technical field
The utility model relates to Optical devices technical fields, and in particular to a kind of digital pcr fluorescence detection device.
Background technique
Digital pcr technology is a kind of nucleic acid molecules absolute quantitation technology as third generation round pcr.Digital pcr passes through
A kind of PCR reaction is divided into many smaller PCR reactions (each small reaction averagely includes being no more than a target nucleic acid molecule)
To improve traditional PCR.Each small reaction comprises about 1 or 0 target nucleic acid molecule, and gives at the end of PCR amplification
Positive or negative binary system is read out, and according to the number and ratio of Poisson distribution principle and positive droplet, you can get it that target nucleic acid divides
The starting copy number or concentration of son.Compared to conventional fluorescent quantitative PCR, digital pcr has outstanding sensitivity, specificity
And accuracy, the fields such as lesion detection, pre-natal diagnosis and unicellular research are had been widely used at present.
At present in industry the digital pcr detecting instrument based on microlayer model technology mainly use drop formation, amplified reaction and
The three-step approach of fluorescence detection realizes the detection of sample, is needed in the actual operation process through a variety of instrument and equipments and is related to drop
Transfer process, risk complicated for operation and there are cross contamination and droplet breakage, so as to cause the accurate of testing result is influenced
Property.
It can easier, efficient digital pcr fluorescence detection device therefore, it is necessary to develop one kind.
Utility model content
In view of this, the present invention provides a kind of digital pcr fluorescence detection device, which can be by microlayer model
Two functions of pcr amplification reaction and fluorescence detection it is highly integrated, realize the augmentation detection in situ of PCR；Whole device can save
It goes droplet transfer present in detection process etc. complicated for operation, shortens sample reaction and detection time, avoid existing and intersect dirt
The risk of dye and droplet breakage, improves the accuracy of testing result.
The utility model provides a kind of digital pcr fluorescence detection device, comprising:
Support base；
Bracket is fixed in the support base；
Objective table, is set to the support base surface, and the objective table includes electronic slide unit, is fixed on the electronic slide unit
On the fixed reaction chamber of sample and temperature-controlling module for regulating and controlling the fixed reaction chamber temperature of the sample, the sample fix
Reaction chamber is for accommodating PCR sample to be measured；The electronic slide unit can be along X, the moving in two dimensional directions of Y-axis；
Optical module, on the bracket, the optical module includes light source portion, optical filter wheel, object lens and CCD figure for setting
As sensor；Wherein, the light beam that the light source portion issues successively is irradiated to institute after the optical filter wheel and the object lens
It states in PCR sample to be measured, the transmitting light that the PCR sample to be measured is issued through irradiation is successively through the object lens and the optical filter wheel
Afterwards, the fluorogenic quantitative detection to the PCR sample to be measured is realized into the ccd image sensor.
Optionally, the optical filter wheel includes multiple filter unit groups, and each filter unit group includes exciting light filter
Mating plate, transmitting light optical filter and dichroscope, the light beam that the light source portion issues successively pass through the exciting light optical filter, described
It is irradiated in the PCR sample to be measured after dichroscope and the object lens；The hair that the PCR sample to be measured is issued through irradiation
Light is penetrated successively after the object lens and the dichroscope and transmitting light optical filter, into the ccd image sensor.
Optionally, the light source portion includes at least two light sources, and the digital pcr fluorescence detection device further includes splicer
Component, the light beam issued by least two light source enter the optical filter wheel after the light combination device assembly.
Optionally, the digital pcr fluorescence detection device further includes stepper motor, the fixed bracket of the stepper motor
On, it is equipped with synchronous belt between the stepper motor and the optical filter wheel, is driven by the rotation of the stepper motor described
Optical filter wheel rotation, to realize the switching between multiple filter unit groups in the optical filter wheel.
Optionally, the fixed reaction chamber of the sample include cavity, be set to the intracorporal temperature-uniforming plate of the chamber and with the cavity
Cover the cover board closed；The fixed reaction chamber of the sample is placed on the electronic slide unit, the temperature-controlling module and the temperature-uniforming plate
Electrical connection is to carry out heat transfer to the temperature-uniforming plate；The cover board includes a transmission region.
Optionally, the ventilating joint of the connection cavity is additionally provided on the cover board, the ventilating joint is for connecting
The air pressure component of peripheral hardware is to regulate and control the intracorporal air pressure of the chamber.
Optionally, the temperature-controlling module includes cooling piece, heat dissipation wind channel, radiator fan, radiator and temperature sensing
Device；The cooling piece is bonded with the temperature-uniforming plate to realize heat transfer；The heat dissipation wind channel is used to provide the air duct of air flowing,
The radiator fan is placed in heat dissipation wind channel to accelerate air to flow；The radiator passes through the heat dissipation wind channel and the heat dissipation
Fan radiates to the cooling piece, and the temperature sensor is used to monitor the temperature of the cooling piece.
Optionally, the electronic slide unit include the bottom plate being cascading in the support base, the first slide plate and
Second slide plate is equipped with the first sliding rail and the first driving motor, to realize first slide plate between the bottom plate and the first slide plate
It is moved along first direction；The second sliding rail and the second driving motor are equipped between first slide plate and second slide plate, with reality
Existing second slide plate moves in a second direction, and the first direction and the second direction are mutually perpendicular to.
Optionally, the ccd image sensor and the optical filter wheel are arranged in parallel on the bracket, the CCD figure
As sensor it is parallel with the optical filter wheel between a right angle gyrator is set, the right-angled corner device is for making by the filter
Right angle turns occur for the transmitting light after mating plate wheel to inject the ccd image sensor.
Optionally, the digital pcr fluorescence detection device is further equipped with digital pcr chip, and the digital pcr chip is flat
It spreads and is fixed in the fixed reaction chamber of the sample, the PCR sample to be measured is placed in the digital pcr chip to be counted
Word PCR reaction.
Optionally, the digital pcr chip includes chip substrate, and the chip substrate is equipped with liquid storage mouth and the storage
Liquid mouth connection first runner, oil storage mouth, second flow channel be connected to the oil storage mouth, drop tile chamber and with the drop put down
Spread the overflow port of chamber connection；For the liquid storage mouth for injecting PCR reaction solution, the oil storage mouth is described for injecting oily phase reagent
Drop tiling chamber is communicated with a mixing runner, and the mixing runner is equipped with mixing entrance far from one end of drop tiling chamber；
The first runner and the second flow channel are intersected in the mixing entrance, and the PCR reaction solution and the oily phase reagent are in institute
It states and is mixed into oral sex remittance generation drop；It tiles the intracavitary pcr amplification reaction for carrying out the drop and detection in the drop.
Optionally, the drop of the digital pcr chip, which tiles, intracavitary is additionally provided with several support columns being intervally arranged.
The support column can be effectively prevented drop tiling chamber and collapse under ambient pressure, improve the entire digital pcr chip
Fastness and impact resistance.
Optionally, a positioning gap is additionally provided on the chip substrate of the digital pcr chip.The locating notch one
Aspect can further prevent the movement of digital pcr chip, on the other hand can effectively prompt the side of putting of digital pcr chip
Position.
Optionally, the light source includes LED, tungsten halogen lamp or laser.
The utility model has the beneficial effects that
(1) digital pcr fluorescence detection device described in the utility model is capable of providing the temperature that PCR sample carries out PCR amplification
Control assembly is realized the augmentation detection in situ of PCR by optical module, can save droplet transfer present in detection process etc.
It is complicated for operation, sample reaction and detection time are shortened, the risk there are cross contamination and droplet breakage is avoided, improves detection knot
The accuracy of fruit.
(2) digital pcr fluorescence detection device described in the utility model is equipped with various light sources, has multi-fluorescence channel, leads to
The automatic switchover for crossing different fluorescence channels realizes multiplex PCR detection, reduces sample requirements；In conjunction with ccd image sensor and
Electronic slide unit can carry out multistation imaging and image procossing to the different fluorescence channels of big detection zone, improve the inspection of product
Degree of testing the speed, precision and automatization level.
(3) digital pcr fluorescence detection device described in the utility model is further equipped with digital pcr chip, can be further real
Now the integrated analysis of the generation of microlayer model, pcr amplification reaction to optical detection is detected.
The advantages of the utility model, will partially illustrate in the following description, and a part is aobvious and easy according to specification
See, or can be known by the implementation of the utility model embodiment.
Detailed description of the invention
Fig. 1 is the structural schematic diagram for the digital pcr fluorescence detection device 100 that an embodiment of the present invention provides；
Fig. 2 is the structural schematic diagram for the electronic slide unit that an embodiment of the present invention provides；
Fig. 3 is the structural representation of sample fixed reaction chamber and temperature-controlling module that an embodiment of the present invention provides
Figure；
Fig. 4 is the structural representation of sample fixed reaction chamber and temperature-controlling module that another embodiment of the utility model provides
Figure；
Fig. 5 is the structural schematic diagram for the optical module that an embodiment of the present invention provides；
Fig. 6 is the light path schematic diagram for the digital pcr fluorescence detection device that an embodiment of the present invention provides；
Fig. 7 is the structural schematic diagram for the digital pcr chip that an embodiment of the present invention provides；
Fig. 8 is the cross section structure schematic diagram for the digital pcr chip that an embodiment of the present invention provides；
Fig. 9 is the structural schematic diagram for the digital pcr chip that another embodiment of the utility model provides.
Specific embodiment
As described below is the preferred embodiment of the utility model embodiment, it is noted that for the general of the art
For logical technical staff, under the premise of not departing from the utility model embodiment principle, several improvements and modifications can also be made,
These improvements and modifications are also considered as the protection scope of the utility model embodiment.
Divide multiple embodiments that the utility model embodiment is further detailed below.Wherein, the utility model is real
It applies example and is not limited to specific embodiment below.Within the scope of the unchanged main rights, implementation can appropriate be changed.
As shown in Figure 1, a digital pcr fluorescence detection device 100 is present embodiments provided, including
Support base 10, bracket 20, objective table 30, optical module 40.
Wherein, the support base 10 can provide a horizontal plane, and the bracket 20, objective table 30 is detachably
Ground is fixed on the horizontal plane；To make the digital pcr device 100 of the entire channel fluorescence detection put in level
Set state.Optionally, the support base 10 is furnished with several foots.The foot is gone back for providing except support
Adjustable height is to realize that support base 10 is horizontal positioned.In the utility model, the support base cross sectional shape can with but
It is not limited to rectangular shape.The cross sectional shape of the support base can also be round, trapezoidal or polygon.
Wherein, the objective table is set to 10 surface of support base, and the objective table 30 includes electronic slide unit 31, fixes
The fixed reaction chamber 32 of sample on the electronic slide unit 31 and the temperature for regulating and controlling fixed 32 temperature of reaction chamber of the sample
Control assembly 33, the fixed reaction chamber 32 of the sample is for accommodating PCR sample to be measured；The electronic slide unit 31 can be along X, Y-axis two
It is mobile to tie up direction；
Wherein, optical module 40 are arranged on the bracket 20, and the optical module 40 includes light source portion 41, optical filter
Wheel 42, object lens 43 and ccd image sensor 44；Wherein, the light beam that the light source portion 41 issues successively passes through the optical filter wheel
42 and the object lens 43 after be irradiated in the PCR sample to be measured, the PCR sample to be measured through irradiation issue transmitting light
Successively after the object lens 43 and the optical filter wheel 42, into the ccd image sensor 44 to realize to described to be measured
The fluorogenic quantitative detection of PCR sample.
Optionally, as shown in Fig. 2, it includes stacking gradually to set that embodiment described in the utility model, which provides the electronic slide unit 31,
Set bottom plate 311, the first slide plate 312 and the second slide plate 313 in the support base 10, the bottom plate 311 and the first slide plate
The first sliding rail 314 and the first driving motor 315 are equipped between 312, to realize that first slide plate 312 is moved along first direction；
The second sliding rail 316 and the second driving motor 317 are equipped between first slide plate 312 and second slide plate 313, to realize
It states the second slide plate 313 to move in a second direction, the first direction and the second direction are mutually perpendicular to.The first direction and
The second direction is any one of X-axis and Y direction.When first direction is X-direction, second direction is Y direction；
When first direction is Y direction, second direction is X-direction.
Wherein, between bottom plate 311 and the first slide plate 312, two close end lateral side regions is leaned on, are equipped with a pair in parallel along first direction
First sliding rail 314, the first slide plate 312 can be moved relative to the bottom plate along first direction by the first sliding rail.By adding
One driving motor 315 may be implemented the first slide plate 312 described in electrical adjustable and move along first direction.Similarly, pass through electric adjustable
Control may be implemented second slide plate 313 and move in a second direction relative to the first slide plate 312.Optionally, first driving
Motor or the second driving motor can be existing servo motor, screw rod stepper motor etc..Pass through the equipment such as host computer or processor
Entirely electronic slide unit moves the instruction electrical adjustable of offer along X-axis and Y direction, to drive the fixed reaction chamber of the sample
32 synchronizing movings.
It originally is in utility model, referring to Fig. 3, the fixed reaction chamber 32 of the sample includes cavity 321, is set to the cavity
Temperature-uniforming plate 322 in 321 and the cover board 323 with the cavity 321 lid conjunction；The fixed reaction chamber 32 of the sample is placed in the electricity
On dynamic slide unit 31, the temperature-controlling module 33 is electrically connected to carry out hot biography to the temperature-uniforming plate 322 with the temperature-uniforming plate 322
It leads；The cover board 323 includes a transmission region 324.
In the present embodiment, can tile PCR sample 325 to be measured on the temperature-uniforming plate 322.Optionally, the temperature-uniforming plate 322
Uniformity of temperature profile with good heat transfer property, on the entire temperature-uniforming plate 322.By being electrically connected with temperature-controlling module 33
It connects, may be used to provide temperature gradient required for pcr amplification reaction.Optionally, the surface of the temperature-uniforming plate 322 is equipped with gold plating
Layer or silvering.The thermal conductivity of temperature-uniforming plate 322 can be improved in the gold plate or silvering.Optionally, the gold plating
Layer or silvering with a thickness of 1-5mm.
Optionally, the ventilating joint 326 of the connection cavity is additionally provided on the cover board 323, as shown in figure 3, described
Ventilating joint is used to connect the air pressure component of peripheral hardware to regulate and control the intracorporal air pressure of the chamber.
In the present embodiment, the lid of the cover board closes the space that the cavity can be made to form a sealing.Optionally, the lid
Sealing ring 327 is additionally provided between plate 323 and the cavity 321.The sealing ring can be fixed in side edge thereof or be fixed on
On cavity edge.The sealing ring is conducive to promote the sealing effect after cover board and cavity cover conjunction.
In the present embodiment, the cavity 321 of the fixed reaction chamber 32 of the sample can be, but not limited to offer for one recessed
The support plate of slot.The bottom of the cavity 321 is the temperature-uniforming plate 322.
As shown in figure 4, the cover board 323 can be connect by rotary shaft 328 with the cavity 321, to realize the lid
Plate is overturn by axis on cavity and realizes folding.When the cover plate lid closes the cavity, between the cover board and the cavity
It is additionally provided with lock.Optionally, the cover board can also be connect with the cavity 321 by other means.
In the present embodiment, the temperature-controlling module 33 can be, but not limited to setting in fixed 32 He of reaction chamber of the sample
Between electronic slide unit 31.Optionally, between the fixed reaction chamber 32 of the sample and the electronic slide unit 31 by setting column into
Row is fixed.It can select, as shown in figure 4, the fixed reaction chamber 32 of the sample has a fixed plate 329, the fixed plate 329 is solid
It is scheduled on the second slide plate of electronic slide unit；It is equipped with column 330 between the fixed reaction chamber 32 of the sample and fixed plate 329, with shape
At an accommodation space 320, temperature-controlling module 33 is set in the accommodation space 320.
Such as Fig. 4, the temperature control group 33 includes cooling piece 331, heat dissipation wind channel 332, radiator fan 333, radiator 334
And temperature sensor；The cooling piece 331 is bonded with the temperature-uniforming plate 322 to realize heat transfer；The heat dissipation wind channel 332 is used for
The air duct of air flowing is provided, the radiator fan 333 is placed in heat dissipation wind channel 332 to accelerate air to flow；The radiator
334 radiate to the cooling piece 331 by the heat dissipation wind channel 332 and the radiator fan 333, the temperature sensor
For monitoring the temperature of the cooling piece.
Optionally, the cooling piece 331 can be, but not limited to include semiconductor chilling plate, and the cooling piece 331 both can be with
Refrigeration can also heat.The temperature sensor can be set in the cooling piece 331 with the temperature of cooling piece described in real-time monitoring.
Optionally, the temperature control group 33 can also realize the PID control to temperature by cooperation host computer.The radiator 334
Effectively cooling piece can be implemented to radiate；The radiator 334 can be more heat-dissipating pipe compositions.
In the present embodiment, the cooling piece 331 is in the one side refrigeration being electrically connected with the temperature-uniforming plate 322, the refrigeration
The heat that the hot end face of piece 331 generates can carry out heat transfer by radiator 334, and pass through heat dissipation wind channel 332 and heat dissipation
333 heat of fan transmits outward, reduces hot end face thermal accumlation, improves cooling rate.The fan speed shelves of the radiator fan
Position can be adjusted in real time, so that entire temperature control group 33 operates normally, and provide the ladder of temperature required for pcr amplification reaction
Degree.
In the present embodiment, see that Fig. 1, the optical module are fixed on the bracket；Wherein, the 44 face institute of object lens
The transmission region of fixed 32 cover board of reaction chamber of sample is stated, the object lens 43 are fixed on the surface of the fixed reaction chamber 32 of sample；?
The object lens 43 are equipped with optical filter wheel 42 far from the side of the fixed reaction chamber 32 of the sample.Optionally, as shown in figure 5, it is described
Optical filter wheel 42 includes multiple filter unit groups 421, and the optical filter wheel 42 is circular pan-like structure, and multiple optical filterings are single
Tuple 421 is distributed in the circular pan-like structure, and using the circular pan-like structure center as center of circle regular array.Often
A filter unit group 421 is a fluorescence channel.For example, filter unit group 421 may include feux rouges light-filtering channels, green light filter
Optical channel, blue light light-filtering channels or near infrared filter channel.
Optionally, the object lens 43 include plane achromatic objective, and the object lens have automatic focusing function.The object lens
With Power focus structure, optical automatic focusing may be implemented；It may be implemented at optical automatic focusing and image sharpening
Reason saves manual focus operation, improves image quality.Optionally, the amplification factor of the object lens is including being 4-20 times.It is preferred that
Ground, the amplification factor of the object lens is including being 4-8 times.
Optionally, the ccd image sensor 44 is arranged in parallel on the bracket 20 with the optical filter wheel 42, institute
It states and is provided with a right angle gyrator 45 between ccd image sensor 44 and the optical filter wheel 42, the right-angled corner device 45 is used
In make the transmitting light right angle turns issued through optical filter wheel 42 to inject the ccd image sensor.Pass through the right-angled corner
Device 45 can carry out 90 ° of corners with optical path, and ccd image sensor is avoided to be fixed on the optical filter wheel vertically, cause to scheme because of CCD
As sensor vertical display causes whole device excessively high and occupied space, therefore, the right-angled corner device 45 is conducive to entirely fill
That sets is integrated, reduces the whole height of digital pcr fluorescence detection device, reduces space hold.
Optionally, the ccd image sensor 44 includes CCD camera；The CCD camera is used to carry out fluorescence channel high
It clearly takes pictures imaging, and imaging results is uploaded to the host computers such as computer and carry out image procossing and analytical calculation.
Digital pcr fluorescence detection device described in the utility model is capable of providing the temperature control that PCR sample carries out PCR amplification
Component processed realizes the augmentation detection in situ of PCR by optical module, can save the behaviour such as droplet transfer present in detection process
Make complexity, shortens sample reaction and detection time, avoid the risk there are cross contamination and droplet breakage, improve testing result
Accuracy.
Optionally, the digital pcr fluorescence detection device 100 further includes stepper motor 46, and the stepper motor 46 is fixed
On the bracket 20, it is equipped with synchronous belt 47 between the stepper motor 46 and the optical filter wheel 42, passes through the stepper motor
46 rotation is to drive the optical filter wheel 42 to rotate, to realize multiple filter unit groups 421 in the optical filter wheel 42
Between switching.
Optionally, the light source portion includes at least two light sources.As shown in figure 5, light source portion 41 includes 411 He of first light source
Second light source 412.
Shown referring to figs. 5 and 6 together, light source portion 41 includes first light source 411 and second light source 412；The digital pcr
Fluorescence detection device further includes light combination device assembly 48, and the light beam issued by least two light source is through the light combination device assembly 48
Enter the optical filter wheel afterwards.The optical filter wheel 42 includes multiple filter unit groups 421, each filter unit group 421
Including exciting light optical filter 422, transmitting light optical filter 423 and dichroscope 424；When light source portion 41 issues light by single source
Shu Shi, the light beam that light source portion 41 issues are irradiated to after successively passing through exciting light optical filter 422, dichroscope 424 and object lens 43
325 in PCR sample to be measured；The transmitting light that the PCR sample 525 to be measured is issued through irradiation is successively through object lens 42 and dichroscope
424 and transmitting light optical filter 423 after, into the ccd image sensor 44.
When light source portion 41 issues light beam by more than two light sources, more than two light sources (such as first light source 411 and the
Two light sources 412) after the single light beam that is first formed after light combination device assembly 48, successively pass through exciting light optical filter 422, dichroic
It is irradiated to 325 in PCR sample to be measured after mirror 424 and object lens 43；The transmitting light that the PCR sample 525 to be measured is issued through irradiation
Successively after object lens 42 and dichroscope 424 and transmitting light optical filter 423, into the ccd image sensor 44.
In the present embodiment, light combination device assembly 48 be can be, but not limited to as dichroscope component, and multiple light sources are by discharging
Position can form single light beam by light combination device assembly 48.
For example, when digital pcr fluorescence detection device described in the utility model has two channels FAM and HEX, the filter
Mating plate wheel can be, but not limited to include two filter unit groups；Wherein, a filter unit group is the channel FAM, the channel FAM
Excitation center wavelength of light is 470nm, and transmitting center wavelength of light is 525nm, and dichroscope cutoff wavelength is 505nm；Another filter
Light unit group is the channel HEX, and the excitation center wavelength of light in the channel HEX is 530nm, and transmitting center wavelength of light is 565nm, two
It is 550nm to Look mirror cutoff wavelength.Exciting light optical filter, transmitting light optical filter and dichroscope in the filter unit group are pressed
According to corresponding parameter setting.
Digital pcr fluorescence detection device described in the utility model can be realized the fluorescence detection of multichannel, can be using electricity
Flowing mode realizes the automatic switchover of different fluorescence channels, and combines ccd image sensor and electronic slide unit to big detection zone
Different fluorescence channels carry out multistation imaging and image procossing, finally calculate concentration of specimens.Described device is greatly improved production
Detection speed, analysis precision and the automatization level of product.
Optionally, the digital pcr fluorescence detection device is while optical detection, the movement process and light of electronic slide unit
The synchronous execution of testing process is learned, imaging is carried out to drop in each station, then splices the image of multistation
Image overlapping region is deleted in analysis, realizes big detection zone, the multistation imaging of high-resolution and image procossing.
Digital pcr fluorescence detection device described in the utility model is further equipped with digital pcr chip, the digital pcr chip
It tiles and is fixed in the fixed reaction chamber of the sample, the PCR sample to be measured is placed in the digital pcr chip to be counted
Word PCR reaction.
Together referring to figs. 7 and 8, another embodiment of the utility model provides a kind of digital pcr chip, the number
Pcr chip includes chip substrate 50, the first runner that the chip substrate is equipped with liquid storage mouth 501, is connected to the liquid storage mouth
502, oil storage mouth 503, the second flow channel 504 being connected to the oil storage mouth, drop tiling chamber 505 and with the drop tile chamber
The overflow port 506 of connection；The liquid storage mouth 501 is used to inject oily phase reagent for injecting PCR reaction solution, the oil storage mouth 503,
Drop tiling chamber 505 is communicated with a mixing runner 507, the mixing runner 507 far from drop tiling chamber 505 one
End is equipped with mixing entrance 508；The first runner 502 and the second flow channel 504 are intersected in the mixing entrance 508, described
PCR reaction solution and the oily phase reagent cross in the mixing entrance 508 and generate drop；The drop tiling chamber 505 in into
The pcr amplification reaction of the row drop and detection.
In the present embodiment, the digital pcr fluorescence detection device is mainly the drop tiling chamber 505 to digital pcr chip
The tiled area at place is detected.Correspondingly, the transmission region of the cover board of the fixed reaction chamber of the sample also mainly wherein exists
The tiled area of drop tiling chamber 505.In drop tiling chamber 505, drop is monolayer distribution, in this way can be more quasi-
True carries out PCR detection to drop.Optionally, settable mutual between the overflow port 506 and drop tiling chamber 505
The third flow channel 509 of connection.
Optionally, the section cross-sectional shape of the liquid storage mouth 501, the oil storage mouth 503 or the overflow port can be
Round or rectangle.The liquid storage mouth, the oil storage mouth or the overflow port can be tubbiness or the slot of inverted cone shape.The liquid storage
Mouth, the oil storage mouth or the overflow port can be, but not limited to offer the through-hole of connection runner in bottom.
Optionally, the bore D of the liquid storage mouth 501 is 3-4mm, depth H 5-6mm；The bore of oil storage mouth 503 is 5-
6mm, depth 5-6mm；The bore of the overflow port 506 is 5-6mm, depth 5-6mm.The cross of the drop tiling chamber 505
Cross-sectional length is 25-35mm, cross-sectional width 10-20mm, is highly 0.08-0.12mm.The drop tiling chamber at least may be used
To accommodate nearly 20,000 drops.Optionally, in digital pcr chip, PCR reaction solution and oil mutually examination is added by providing normal pressure
Agent, the oily phase reagent that the PCR reaction solution and the second flow channel 504 for flowing into the first runner 502 flow into is in the mixing
Entrance 508 crosses, effect high throughput and big rule of the PCR reaction solution sample in runner using the Osima jacoti, Osima excavata of oily phase reagent
The oily packet drop of the generation dispersion of mould.Optionally, the positive pressure value is 30-50kPa.
Optionally, the diameter for the drop that the digital pcr chip generates is 0.05-0.1mm.Optionally, described to be mixed into
The pore size of mouth 508 is 0.05-0.1mm.
Optionally, the drop of the digital pcr chip, which tiles, intracavitary is additionally provided with several support columns being intervally arranged.
The support column can be effectively prevented drop tiling chamber and collapse under ambient pressure, improve the entire digital pcr chip
Fastness and impact resistance.
Optionally, it is additionally provided with a positioning gap 510 on the chip substrate of the digital pcr chip, referring to Fig. 9.Institute
On the one hand the movement of digital pcr chip can be further prevented by stating locating notch, play positioning action, on the other hand can be effective
Prompt digital pcr chip put orientation and, it is ensured that the tiled area of chip faces the transmission region of the cover board.
Further, optionally, the end surface of the chip substrate 50 of the digital pcr chip is additionally provided with marked area
Domain 511.The mark region can be used for recording the bar code and version information of sample, to be tracked and to be chased after to test sample
It traces back.
In the present embodiment, the material of the chip substrate is light-transmitting materials.The drop tiling chamber of the digital pcr chip
Top and bottom are plane.The digital pcr chip is laid on the temperature-uniforming plate in the fixed reaction chamber of the sample.The number
Word pcr chip expands temperature ginseng in the fixed reaction chamber of the sample, through the PCR program in the fixed reaction chamber of setting sample
Number, may be implemented PCR amplification, and under the detection of optical module, realize the original position PCR augmentation detection.
Further, by the ventilating joint on the cover board of the fixed reaction chamber of the sample, make in the fixed reaction chamber of sample
Equipped with the cleaning air pressure for being greater than 100kPa, can be effectively prevented in the liquid storage mouth, oil storage mouth or overflow port of the digital pcr chip
Liquid volatilization during heating.
In the present embodiment, after the digital pcr fluorescence detection device is equipped with digital pcr chip, it may be implemented to be measured
The generation of the microlayer model of PCR sample, the integrated analysis of pcr amplification reaction to optical detection detect；It is examined compared to traditional PCR
Survey device, digital pcr fluorescence detection device described in the utility model will be more efficient, more portable and more automate；And due to
Without processes such as other transfers during the generation of microlayer model, three of pcr amplification reaction to optical detection, avoid intersecting
The risk of pollution and droplet breakage, therefore, the detection data of the utility model digital pcr fluorescence detection device is more accurate can
It leans on.
Above-described embodiments merely represent several embodiments of the utility model, the description thereof is more specific and detailed,
But it should not be understood as limiting the scope of the patent of the utility model.It should be pointed out that for the common of this field
For technical staff, without departing from the concept of the premise utility, various modifications and improvements can be made, these all belong to
In the protection scope of the utility model.Therefore, the protection scope of the utility model should be determined by the appended claims.
Claims (10)
1. a kind of digital pcr fluorescence detection device characterized by comprising
Support base；
Bracket is fixed in the support base；
Objective table, is set to the support base surface, and the objective table includes electronic slide unit, is fixed on the electronic slide unit
The fixed reaction chamber of sample and the temperature-controlling module for regulating and controlling the fixed reaction chamber temperature of the sample, the fixed reaction of the sample
Chamber is for accommodating PCR sample to be measured；The electronic slide unit can be along X, the moving in two dimensional directions of Y-axis；
Optical module, on the bracket, the optical module includes that light source portion, optical filter wheel, object lens and ccd image pass for setting
Sensor；Wherein, the light beam that the light source portion issues successively be irradiated to after the optical filter wheel and the object lens it is described to
It surveys in PCR sample, the PCR sample to be measured is irradiated the transmitting light issued successively after the object lens and the optical filter wheel,
The fluorogenic quantitative detection to the PCR sample to be measured is realized into the ccd image sensor.
2. digital pcr fluorescence detection device as described in claim 1, which is characterized in that the optical filter wheel includes multiple filters
Light unit group, each filter unit group include exciting light optical filter, transmitting light optical filter and dichroscope, the light source portion
The light beam of sending is successively irradiated to after the exciting light optical filter, the dichroscope and the object lens described to be measured
In PCR sample；The transmitting light that the PCR sample to be measured is issued through irradiation is successively through the object lens and the dichroscope and transmitting
After light optical filter, into the ccd image sensor.
3. digital pcr fluorescence detection device as described in claim 1, which is characterized in that the light source portion includes at least two
Light source, the digital pcr fluorescence detection device further include light combination device assembly, and the light beam issued by least two light source is through institute
Enter the optical filter wheel after stating light combination device assembly.
4. digital pcr fluorescence detection device as described in claim 1, which is characterized in that the digital pcr fluorescence detection device
Further include stepper motor, on the fixed bracket of the stepper motor, is equipped between the stepper motor and the optical filter wheel
Synchronous belt drives the optical filter wheel to rotate, to realize in the optical filter wheel by the rotation of the stepper motor
Switching between multiple filter unit groups.
5. digital pcr fluorescence detection device as described in claim 1, which is characterized in that the sample fixes reaction chamber and includes
Cavity, the cover board set on the intracorporal temperature-uniforming plate of the chamber and with cavity cover conjunction；The fixed reaction chamber of the sample is placed in described
On electronic slide unit, the temperature-controlling module is electrically connected to carry out heat transfer to the temperature-uniforming plate with the temperature-uniforming plate；The lid
Plate includes a transmission region；The ventilating joint of the connection cavity is additionally provided on the cover board, the ventilating joint is for connecting
The air pressure component of peripheral hardware is connect to regulate and control the intracorporal air pressure of the chamber.
6. digital pcr fluorescence detection device as claimed in claim 5, which is characterized in that the temperature-controlling module includes system
Cold, heat dissipation wind channel, radiator fan, radiator and temperature sensor；The cooling piece is bonded with the temperature-uniforming plate to realize heat
Conduction；The heat dissipation wind channel is used to provide the air duct of air flowing, and the radiator fan is placed in heat dissipation wind channel to accelerate air
Flowing；The radiator radiates to the cooling piece by the heat dissipation wind channel and the radiator fan, and the temperature passes
Sensor is used to monitor the temperature of the cooling piece.
7. digital pcr fluorescence detection device as described in claim 1, which is characterized in that the electronic slide unit includes successively layer
Folded bottom plate, the first slide plate and the second slide plate being arranged in the support base is equipped with the between the bottom plate and the first slide plate
One sliding rail and the first driving motor, to realize that first slide plate is moved along first direction；First slide plate and described second
The second sliding rail and the second driving motor, to realize that second slide plate moves in a second direction, described first are equipped between slide plate
Direction and the second direction are mutually perpendicular to.
8. digital pcr fluorescence detection device as described in claim 1, which is characterized in that the ccd image sensor with it is described
Optical filter wheel is arranged in parallel on the bracket, is arranged always between the ccd image sensor is parallel with the optical filter wheel
Angle gyrator, the right-angled corner device is for making the transmitting light after the optical filter wheel that right angle turns occur to inject
The ccd image sensor.
9. digital pcr fluorescence detection device as described in claim 1, which is characterized in that the digital pcr fluorescence detection device
It is further equipped with digital pcr chip, the digital pcr chip tiles and is fixed in the fixed reaction chamber of the sample, described to be measured
PCR sample is placed in the digital pcr chip to carry out digital pcr reaction.
10. digital pcr fluorescence detection device as claimed in claim 9, which is characterized in that the digital pcr chip includes core
Plate base, the chip substrate are equipped with liquid storage mouth, the first runner being connected to the liquid storage mouth, oil storage mouth and the oil storage
Second flow channel, drop tiling chamber and the overflow port being connected to drop tiling chamber of mouth connection；The liquid storage mouth is for injecting
PCR reaction solution, the oil storage mouth are communicated with a mixing runner, the mixing for injecting oily phase reagent, the drop tiling chamber
Runner is equipped with mixing entrance far from one end of drop tiling chamber；The first runner and the second flow channel are intersected in described
It mixes entrance, the PCR reaction solution and the oily phase reagent and is mixed into oral sex remittance generation drop described；It tiles in the drop
The intracavitary pcr amplification reaction for carrying out the drop and detection.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110699249A (en) * | 2019-11-01 | 2020-01-17 | 宁波胤瑞生物医学仪器有限责任公司 | Nucleic acid amplification instrument |
| CN110975960A (en) * | 2019-12-24 | 2020-04-10 | 中国科学院长春光学精密机械与物理研究所 | Composite PCR system with dPCR and qPCR functions and control method |
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| CN111394219A (en) * | 2020-03-20 | 2020-07-10 | 中国科学院长春光学精密机械与物理研究所 | Integrated digital PCR system |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110699249A (en) * | 2019-11-01 | 2020-01-17 | 宁波胤瑞生物医学仪器有限责任公司 | Nucleic acid amplification instrument |
| CN110699249B (en) * | 2019-11-01 | 2021-08-06 | 宁波胤瑞生物医学仪器有限责任公司 | Nucleic acid amplification instrument |
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| CN111004721A (en) * | 2019-12-24 | 2020-04-14 | 南通大学 | Ultrathin visual temperature control device suitable for microscope online in-situ observation |
| CN110975960B (en) * | 2019-12-24 | 2022-05-24 | 中国科学院长春光学精密机械与物理研究所 | Composite PCR system with dPCR and qPCR functions and control method |
| CN111394219A (en) * | 2020-03-20 | 2020-07-10 | 中国科学院长春光学精密机械与物理研究所 | Integrated digital PCR system |
| CN111748463A (en) * | 2020-08-07 | 2020-10-09 | 天津智善生物科技有限公司 | Amplification device applied to nucleic acid detection and detection method based on amplification device |
| CN111909842A (en) * | 2020-08-25 | 2020-11-10 | 重庆大学 | Integrated digital PCR system and use method thereof |
| CN112595668A (en) * | 2020-11-27 | 2021-04-02 | 南京溯远基因科技有限公司 | Rapid multi-fluorescence real-time quantitative PCR instrument |
| CN113219611A (en) * | 2021-04-26 | 2021-08-06 | 宁波胤瑞生物医学仪器有限责任公司 | Electric conversion device for optical filter |
| CN113970535A (en) * | 2021-10-18 | 2022-01-25 | 广州市微米生物科技有限公司 | Detector and using method thereof |
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