CN107860701A - Droplet type sample introduction fluoroscopic examination control system - Google Patents
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- CN107860701A CN107860701A CN201711407907.XA CN201711407907A CN107860701A CN 107860701 A CN107860701 A CN 107860701A CN 201711407907 A CN201711407907 A CN 201711407907A CN 107860701 A CN107860701 A CN 107860701A
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- 238000001514 detection method Methods 0.000 claims abstract description 118
- 238000012360 testing method Methods 0.000 claims abstract description 20
- 238000001917 fluorescence detection Methods 0.000 claims abstract description 9
- 239000000523 sample Substances 0.000 claims description 67
- 230000003287 optical effect Effects 0.000 claims description 22
- 239000007788 liquid Substances 0.000 claims description 11
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- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 claims description 3
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 claims description 3
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims description 3
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N2015/1024—Counting particles by non-optical means
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- General Physics & Mathematics (AREA)
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- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The present invention relates to a kind of droplet type sample introduction fluoroscopic examination control system, including sample introduction module, droplet detection module and master control system, the sample introduction module is used to extract droplet sample and push the droplet to droplet detection module；The droplet detection module is used to carry out signal detection to the droplet to obtain signal detection data；The master control system is electrically connected to the sample introduction module and the droplet detection module, for controlling sample introduction module to extract droplet sample and pushing the droplet to droplet detection module, control droplet detection module to carry out signal detection, obtain testing result and the testing result data are sent to computer system and handled and analyzed.The present invention uses droplet type fluorescence detection method, compared with using more orifice-plate type or chip type detection method on domestic market, has the advantages that single reactor volume is smaller, and flux is big, and cost is low, and precision is high.
Description
Technical field
The present invention relates to droplet type technical field of fluorescence detection, is controlled more particularly to a kind of droplet type sample introduction fluoroscopic examination
System.
Background technology
Digital pcr (Polymerase Chain Reaction PCRs) is although technology proposes so far only
More than ten years, but due to its unique technical advantage and application prospect so that its industrialized development is quite rapid.It is so far
Only, including Fluidigm, Life Tech, Bio-Rad and Raindance companies (being purchased by Bio-Rad at the beginning of 2017)
Deng Ji companies have released one after another digital pcr product, and are applied to single cell analysis, early diagnosis of cancer and pre-natal diagnosis
Etc. research field.Numbers above PCR, Inc. is offshore company, and Fluidigm and Life Tech companies mainly use chip
Formula is detected, and it detects flux finite, and testing cost is higher；Bio-Rad and Raindance companies are examined using droplet
Survey, its flux is high, and detection sensitivity is high, and cost is low, and market acceptance is higher.
And domestic market is mainly orifice-plate type quantitative fluorescent PCR, detection all fluorescence intensities of amplification procedure are, it is necessary to and standard
Curve is to mark, it is impossible to which absolute quantitation, precision are low.
The content of the invention
Based on this, it is necessary to provide a kind of droplet type sample introduction fluoroscopic examination control system, it is intended to make the quantity of droplet sample
Can be by accurate statistics and analysis.
A kind of droplet type sample introduction fluoroscopic examination control system, including sample introduction module, droplet detection module and main control system
System, the sample introduction module are used to extract droplet sample and push the droplet to droplet detection module；The droplet detection module
For carrying out signal detection to the droplet to obtain signal detection data；The master control system enters original mold described in being electrically connected to
Block and the droplet detection module, mould is detected for controlling sample introduction module to extract droplet sample and pushing the droplet to droplet
Block, control droplet detection module carry out signal detection, obtain testing result and the testing result data are sent into computer
System is handled and analyzed.
In one of the embodiments, the sample introduction module includes fluid path control system, and the fluid path control system includes
Peristaltic pump, syringe pump, valve and motor, the fluid path control system are controlled by the master control system, in peristaltic pump, note
Penetrating under the mutual cooperation of pump, valve and motor has detergent line, extracts droplet, the work(of push droplet to droplet detection module
Energy.
In one of the embodiments, the motor is stepper motor or DC brushless motor.
In one of the embodiments, the droplet detection module includes fluorescence detection optical path system, the fluoroscopic examination
The fluorescence signal that light path system is used to detect droplet detects data to obtain fluorescence signal.
In one of the embodiments, the droplet detection module includes scattering light light path system, the scattering light
The scattered light signal that light path system is used to detect droplet detects data to obtain scattered light signal.
In one of the embodiments, the droplet detection module includes detection chip, light source, optical filter, detection
Device, signal detection plate and light path system structure, the detection chip offer sense channel, had in the sense channel to micro-
Drip the test point detected；The light irradiation that the light source is sent makes biology in droplet on test point, for irradiating droplet
Sample is inspired fluorescence signal and/or droplet is inspired scattering light under spheric reflection effect；The optical filter
For being filtered to the fluorescence signal of different wave length, to obtain the fluorescence signal of specific wavelength；The detector is used for glimmering
Optical signal and/or scattered light signal are detected, and obtain fluorescence signal detection data and/or scattered light signal detection data；Institute
State signal detection plate and be used for fluorescence signal detection data and/or scattered light signal detection data that acquisition testing obtains；The light
Road system architecture, for fixing and/or adjusting the optical element in the droplet detection module, so that the droplet detection module
Form complete light path system.
In one of the embodiments, the detection chip and the quantity of detector are one or more；The detection core
The species of piece and detector is one or more.
In one of the embodiments, the detection chip is PDMS chips, COC chips, COP chips, PMMA chips, PC
Chip, glass-chip, capillary or flow cell；The light source is solid state laser light source, gas laser light source, semiconductor
Laser light source or LED light source；The detector is detected using PMT or PD sensors are detected.
In one of the embodiments, in addition to state detection module, sample introduction needle detection module, radiating module and power supply mould
Block, the state detection module are used to detect whole system equipment including detecting 96 orifice plate placement locations, waste liquid liquid level, oil
Liquid level, reagent door close situation, instrument door closes situation；The sample introduction needle detection module is used for the exception for detecting sample introduction needle
Situation；The radiating module is used to be radiated for whole system, with each instrument well-ventilated in guarantee system, stable；
The power module is used to provide D.C. regulated power supply for whole system.
In one of the embodiments, the radiating module is radiated using blower fan exhausting heat radiation or cooling piece；The electricity
Source module input is civil power, is exported as 24V DC voltages.
Above-mentioned droplet type sample introduction fluoroscopic examination control system, negative and positive can be accurately identified using droplet type fluorescence detection method
The number and droplet total number of droplet, with using more orifice-plate type and chip type detection method phase on domestic market
Than the present invention has the advantages that single reactor volume is smaller, and flux is big, and cost is low, and precision is high.
Brief description of the drawings
Fig. 1 is droplet type sample introduction fluoroscopic examination control system module map provided in an embodiment of the present invention.
Embodiment
Referring to Fig. 1, in one embodiment, there is provided a kind of droplet type sample introduction fluoroscopic examination control system, including enter
Egf block 10, state detection module 20, sample introduction needle detection module 30, droplet detection module 40, radiating module 50, power module
60th, master control system 70 and computer system 80.
The sample introduction module 10 is used to extract droplet sample and push the droplet to droplet detection module 40.In this embodiment party
In formula, the sample introduction module 10 includes peristaltic pump 11, syringe pump 12, valve 13 and motor 14.The syringe pump 12 is used to take out
Droplet sample is taken, and pushes the droplet sample to droplet detection module 40；The peristaltic pump 11 is used for waste liquid (after detection
Droplet sample) waste liquid tank is delivered to, and runner is rinsed；The valve 13 is used to control peristaltic pump 11 and syringe pump 12
System, to adjust the work schedule of peristaltic pump 11 and syringe pump 12, droplet push is carried out in the predetermined time or runner rinses；Institute
Stating motor 14 drives the syringe pump 12 and the valve 13 to act, to complete to extract the function of droplet and droplet sample introduction.
The sample introduction module 10 also includes drive control circuit 15, and the drive control circuit 15 can control motor 14 to drive institute
State syringe pump 12 and corresponding action is completed in the action of the valve 13.In the present embodiment, the motor 14 is stepping electricity
Machine or DC brushless motor.
The motor 14, syringe pump 12, valve 13 and peristaltic pump 11 form the fluid path control system of the sample introduction module 10
System, being cooperated between the syringe pump 12, valve 13 and peristaltic pump 11 can realize under the control of the master control system 70
Detergent line, extract droplet sample, push droplet to the functions such as droplet detection module 40.
Specifically, under the driving of motor 14, it is corresponding that the syringe pump 12 is moved to orifice plate (such as 96 orifice plates) place
Hole position extract droplet sample, be moved to droplet detection module 40 afterwards, it is then micro- by what is be drawn under the control of valve 13
Drop sample pushes to droplet detection module 40, after droplet detection module 40 is to the droplet pattern detection, the wriggling
Waste liquid after detection is delivered to waste liquid tank under the control of valve 13 and runner is rinsed by pump 11.
The state detection module 20 whether there is abnormal feelings for detecting the droplet type sample introduction fluoroscopic examination control system
Condition, if detecting exception, alarm is sent, in the present embodiment, the abnormal conditions are not placed on including orifice plate
Precalculated position, waste liquid liquid level is too high, oil level is too low, reagent door does not close, instrument door does not close.Specifically, the state
Detection module 20 can use optical detecting module (such as optocoupler), electrical detection module (such as voltage, resistance), mechanical inspection
Module (such as limited post, limited block) etc. is surveyed, the present embodiment does not limit.
The sample introduction needle detection module 30 whether there is abnormal conditions for detecting sample introduction needle, if detecting that sample introduction needle is different
Often, then this detection of the mark of master control system 70 is invalid, that is, the data detected are invalid data, in the present embodiment, are entered
Sample pin abnormal conditions are withstood including sample introduction needle by hard thing, sample introduction needle does not pierce through 96 orifice plate sealers, sample introduction needle is blocked etc..Specifically
Ground, the sample introduction needle detection module 30 can also use the sensor of any type, no longer repeat one by one.
The droplet detection module 40 is used to carry out signal detection to the droplet to obtain signal detection data, the droplet
Detection module 40 includes detection chip, light source 41, optical filter, detector 42, signal acquiring board 43 and light path system structure,
Sense channel is offered in the detection chip, the test point detected to droplet is provided with the sense channel, it is described micro-
Drop sample, which is pushed at the test point to sense channel, to be detected, and in the present embodiment, the detection chip can be
PDMS chips, COC chips, COP chips, PMMA chips, PC chips, glass-chip, capillary or flow cell；The light source 41
The biological sample in droplet is inspired the fluorescence signal of different wave length for irradiating droplet and/or make droplet in spheric reflection
Scattering light is sent under effect, in the present embodiment, the light source 41 can be solid state laser, gas laser, semiconductor
Laser or LED light source etc.；The optical filter is used to filter fluorescence signal, to select the fluorescence of specific wavelength to believe
Number pass through；The detector 42 is used to detect the fluorescence signal and/or scattered light signal of specific wavelength, in this embodiment party
In formula, the detector 42 is detected using PMT or the detection of PD sensors, and a variety of detectors 42 can be set to detect a variety of spies
The fluorescence signal of standing wave length；The signal acquiring board 43 is used to gather the detection data after detecting droplet, including fluorescence
Signal detection data and/or scattered light signal detection data, in the present embodiment, the main control chip of the signal acquiring board 43
Can be FPGA, DSP or ARM；The light path system structure includes fixed structure and adjustment structure, and the fixed structure is used for solid
The position of optical element in the fixed droplet detection module 40, to complete building for whole light path system；The adjustment structure is used
In the position for adjusting optical element in light path system, include the position of optical filter, detection chip.In the present embodiment,
The transmission means of optical signal can be transmitted by air in the light path system structure, can also be transmitted using optical fiber.
The radiating module 50 is used to be radiated for whole system, steady with each instrument well-ventilated, operation in guarantee system
Fixed, in the present embodiment, the radiating module 50 is radiated using blower fan exhausting heat radiation or cooling piece.
The power module 60 is used to provide D.C. regulated power supply for whole system, in the present embodiment, the power supply mould
The input of block 60 is civil power, is exported as 24V DC voltages.
Operation of the master control system 70 for the whole system including controlling sample introduction module 10 and droplet detection module 40,
And the testing result data are sent to computer system 80 and is handled and is analyzed, in the present embodiment, the master control
System 70 processed is master control borad, and its main control chip uses FPGA, DSP or ARM.
The computer system 80 is connected with master control system 70 by USB or serial ports, for controlling the behaviour of whole system
Make, read fluorescence signal detection data and scattered light signal detection data that signal acquiring board 43 is gathered, pass through Algorithm Analysis
The number of negative and positive droplet, and handle correlated results.
Further, droplet type sample introduction fluoroscopic examination control system provided by the present invention includes fluorescence detection optical path system
With scattering light light path system, the fluorescence detection optical path system is used to detect the fluorescence signal of droplet to obtain fluoroscopic examination
Data, the fluoroscopic examination data are used for the number for counting negative and positive droplet；The scattering light light path system is micro- for detecting
To obtain scattering light detection data, the scattering light detection data is used to count droplet total number the scattered light signal of drop.
In fluorescence detection optical path system, droplet is pushed the sense channel to detection chip by sample introduction module 10
It is interior, when droplet is by test point in sense channel, after the laser irradiation that the droplet is sent through laser, the biology in droplet
Sample is inspired the fluorescence signal of different wave length, and the fluorescence signal selects the glimmering of specific wavelength after optical filter filters
Optical signal, the fluorescence signal of this specific wavelength is detected by PMT or PD sensors are detected, and the signal acquiring board 43 gathers
The fluorescence signal detection data detected, and fluorescence signal detection data are sent to computer system and is handled and is analyzed.
Further, in the present embodiment, the detection frequency of droplet can be controlled in the range of 100-10000Hz；Sun
Property droplet fluorescence intensity is stronger, is an oscillogram for having peak value quickly through signal during test point；Negative droplet fluorescence model
It is very faint, droplet signal amplitude very little.Positive droplet can accurately be differentiated by algorithm by delimiting amplitude and width.
In light light path system is scattered, when droplet passes through detection chip, laser sends laser and is irradiated to droplet,
Under droplet spheric reflection effect, the light on droplet is irradiated to centered on droplet shadow surface to space scattering, scattering light is formed, takes
Specific light path system structure is built to collect the scattering light in some region, this scattering light is detected by PMT or PD sensors enter
Row detection, the scattered light signal detection data that the acquisition testing of signal acquiring board 43 arrives, and the scattered light signal is detected into data
Transmission is handled and analyzed to computer system, accurately to calculate total droplet number, differentiates the size of droplet, distinguish droplet and
Gas column, and the accuracy of testing result can be provided by the real time contrast of algorithm processing.
It is appreciated that droplet type sample introduction fluoroscopic examination control system of the present invention can be multiple according to real needs setting
Sense channel, such as the heavy sense channels of 2-10, its detection mode and original can be set according to the species of biological reagent mark
Manage same as above.
The droplet type sample introduction fluoroscopic examination control system that the embodiment of the present invention is provided has the advantages that：
1st, the system can accurately identify negative and positive droplet number and droplet total number；
2nd, the sampling rate is high, and single droplet sampling number is more, and accuracy rate is high；
3rd, the system uses droplet type detection mode, and compared with chip type detection mode, single reactor volume is smaller, leads to
Amount is big, and cost is low, and precision is high；
4th, the system controls pipeline capable of washing to avoid polluting by fluid path, and detection chip may be reused, and reduces inspection
Survey cost.
5th, scattering light light path system can aid in detecting quantity, the size of droplet of droplet, distinguish droplet and gas column,
The accuracy of testing result can be provided by real time contrast's processing of algorithm.
Each technical characteristic of embodiment described above can be combined arbitrarily, to make description succinct, not to above-mentioned reality
Apply all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, the scope that this specification is recorded all is considered to be.
Embodiment described above only expresses the several embodiments of the present invention, and its description is more specific and detailed, but simultaneously
Can not therefore it be construed as limiting the scope of the patent.It should be pointed out that come for one of ordinary skill in the art
Say, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the protection of the present invention
Scope.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (10)
- A kind of 1. droplet type sample introduction fluoroscopic examination control system, it is characterised in that including sample introduction module, droplet detection module and Master control system,The sample introduction module is used to extract droplet sample and push the droplet to droplet detection module；The droplet detection module is used to carry out signal detection to the droplet to obtain signal detection data；The master control system is electrically connected to the sample introduction module and the droplet detection module, for controlling sample introduction module to take out Take droplet sample and push the droplet to droplet detection module, control droplet detection module and carry out signal detection, detected As a result and the testing result data are sent into computer system to be handled and analyzed.
- 2. droplet type sample introduction fluoroscopic examination control system according to claim 1, it is characterised in that the sample introduction module bag Fluid path control system is included, the fluid path control system includes peristaltic pump, syringe pump, valve and motor, the fluid path control system System is controlled by the master control system, has cleaning pipe under the mutual cooperation of peristaltic pump, syringe pump, valve and motor Road, extract droplet, the function of push droplet to droplet detection module.
- 3. droplet type sample introduction fluoroscopic examination control system according to claim 2, it is characterised in that the motor is Stepper motor or DC brushless motor.
- 4. droplet type sample introduction fluoroscopic examination control system according to claim 1, it is characterised in that the droplet detects mould Block includes fluorescence detection optical path system, and the fluorescence signal that the fluorescence detection optical path system is used to detect droplet is believed with obtaining fluorescence Number detection data.
- 5. droplet type sample introduction fluoroscopic examination control system according to claim 1, it is characterised in that the droplet detects mould Block includes scattering light light path system, described to scatter light light path system for detecting the scattered light signal of droplet to obtain Scattered light signal detects data.
- 6. droplet type sample introduction fluoroscopic examination control system according to claim 1, it is characterised in that the droplet detects mould Block includes detection chip, light source, optical filter, detector, signal detection plate and light path system structure,The detection chip offers sense channel, has the test point detected to droplet in the sense channel；The light irradiation that the light source is sent makes the biological sample in droplet be inspired fluorescence on test point, for irradiating droplet Signal and/or make droplet spheric reflection effect under be inspired scattering light；The optical filter is used to filter the fluorescence signal of different wave length, to obtain the fluorescence signal of specific wavelength；The detector be used for fluorescence signal and/or scattered light signal are detected, obtain fluorescence signal detection data and/or Scattered light signal detects data；The signal detection plate is used for the fluorescence signal detection data and/or scattered light signal detection data that acquisition testing obtains；The light path system structure, for fixing and/or adjusting the optical element in the droplet detection module, so that described micro- Drop detection module forms complete light path system.
- 7. droplet type sample introduction fluoroscopic examination control system according to claim 6, it is characterised in that the detection chip and The quantity of detector is one or more；The detection chip and the species of detector are one or more.
- 8. droplet type sample introduction fluoroscopic examination control system according to claim 6, it is characterised in thatThe detection chip is PDMS chips, COC chips, COP chips, PMMA chips, PC chips, glass-chip, capillary Or flow cell；The light source is solid state laser light source, gas laser light source, semiconductor laser light source or LED light source；The detector is detected using PMT or PD sensors are detected.
- 9. droplet type sample introduction fluoroscopic examination control system according to claim 1, it is characterised in that also including state-detection Module, sample introduction needle detection module, radiating module and power module,The state detection module be used for detect whole system equipment include detection 96 orifice plate placement locations, waste liquid liquid level, Oil level height, reagent door closes situation, instrument door closes situation；The sample introduction needle detection module is used for the abnormal conditions for detecting sample introduction needle；The radiating module is used to be radiated for whole system, with each instrument well-ventilated in guarantee system, stable；The power module is used to provide D.C. regulated power supply for whole system.
- 10. droplet type sample introduction fluoroscopic examination control system according to claim 1, it is characterised in thatThe radiating module is radiated using blower fan exhausting heat radiation or cooling piece；The power module input is civil power, is exported as 24V DC voltages.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114262733A (en) * | 2022-01-10 | 2022-04-01 | 深圳麦科田生物医疗技术股份有限公司 | Micro-drop digital PCR (polymerase chain reaction) fluorescent signal processing method |
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