CN101403742B - Method for dielectric characterization of micro-nano biological particle by optoelectronic forceps - Google Patents
Method for dielectric characterization of micro-nano biological particle by optoelectronic forceps Download PDFInfo
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- CN101403742B CN101403742B CN2008101950862A CN200810195086A CN101403742B CN 101403742 B CN101403742 B CN 101403742B CN 2008101950862 A CN2008101950862 A CN 2008101950862A CN 200810195086 A CN200810195086 A CN 200810195086A CN 101403742 B CN101403742 B CN 101403742B
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Abstract
The invention provides a method used for carrying out dielectric characterization to micro-nanometer biological particles by utilizing photoelectric forceps, comprising the steps as follows: the movement speed of an optical pattern is gradually changed so as to lead the particle to reach the maximum synchronous speed before out-of-step; and by being combined with the frequency adjustment of an excitation signal, the maximum synchronous speed curve of the biological particle in a certain frequency range of the excitation signal is measured, thus completing the dielectric characterization of the particle. The dielectric characterization method provided by the invention sufficiently utilizes the flexibility advantage of the photoelectric forceps and avoids preparing a complex physical entity electrode array on the chip simultaneously, is better than the existing dielectric characterization method of the micro-nanometer particle on the aspects of cost, function and performance, and provides an extremely important means for the leaping development of the biomedical detection field.
Description
Technical field
The present invention be a kind of photoelectricity micro-fluidic device that uses to the method that micro-nano biomone characterizes and differentiates, relate to micro-fluidic field, particularly micro-fluidic biological medical science chip field.
Background technology
The dielectric property of micro-nano biomone and its structure and chemical composition are closely related, can be used as " fingerprint " of demarcating micro-nano biomone particular type.The process of obtaining the particle dielectric property is called dielectric characterization.Electricity rotation dielectrophoresis (electrorotation dielectrophoresis; ROT-DEP) be exactly technology that is used for the biomone dielectric characterization wherein; It utilizes the alternating voltage signal of several outs of phase on electrorotation chip, to form rotating electric field, and fine particle just can rotate under the effect of rotating electric field.Biomone is because of the difference of its dielectric properties, and the rotation that is produced response is also different.Therefore, utilize these characteristics just can realize dielectric characterization to biomone, this feasible micro-sick cell is identified from a large amount of normal cells becomes possibility.Obtain the dielectric property method of biological particle based on electricity rotation dielectrophoresis technology; Owing to have non-destructive, implement simply, satisfy the Touchless manipulation demand; And characterize chip and have the little integrated level advantages of higher of volume; Become the important technology that enables that present realization biological particle characterizes diagnosis, just become a kind of important method of major disease diagnosis.In addition; Conventional dielectrophoresis (conventional dielectrophoresis; CDEP) and row ripple dielectrophoresis (travelling-wave dielectrophoresis; TwDEP) the dielectrophoresis phenomenon of these two kinds of biological particles also is used to judge qualitatively the dielectric property of biomone once in a while, but needs to make complicated microelectrode, and the extensive degree of application is not as good as electricity rotation dielectrophoresis.On the whole; There is following point in research field at the dielectric characterization chip at present: one, aspect the economy of making the dielectric characterization chip: present electricity rotation dielectrophoresis test chip, and major part has only the simple electrode structure that produces electricity rotation dielectrophoresis, has ignored the sample pre-treatments function; And some integrated chips of pretreatment function such as sample feeding, separation; Owing to need to realize complex manipulation, thereby need design and make corresponding complicated electrode structure that manufacturing process is complicated to biomone; Cost of manufacture is very high, is not suitable for making the disposable detection chip of a large amount of jettisonables that are used for medical diagnosis.Two, aspect the function implementation of the dielectric characterization of biomone: present chip realizes that dielectric characterization depends on the microelectrode array with given shape more; And these electrod-arrays lack flexibility; Cause under condition of different, needing to make different microelectrode arrays; Increased the difficulty and the loaded down with trivial details Cheng Du that make and operate greatly, be difficult to promote the use of.
In recent years, utilization photoelectricity micro-fluidic device is handled micro-nano biomone and is become a kind of new microscopic particle manipulation technology.This principle based on photoelectric flexible operated tool is following: this photoelectricity micro-fluidic device is three layers of hamburger structure; The liquid that contains sample particles is positioned between the glass (upper strata) and photoconductive layer (lower floor) that is coated with ITO (indium-tin oxide) film, and wherein photoconductive layer is deposited on the ito glass of lower floor.Conducting material has high electrical resistance unglazed according under the situation; And when accepting illumination; The area pellucida photoproduction carrier concentration improves rapidly and makes its local conductivity improve several magnitude rapidly; Cause the difference of light and shade district fluid layer dividing potential drop, form inhomogeneous field in the space and produced " light-induction dielectrophoresis " phenomenon.So if with an aperture particle circle is lived, particle is trapped within the aperture by the effect of light-induction dielectrophoresis power so, particle will be with the moving of aperture, and this method of light pattern manipulation of particles of utilizing is called photoelectricity tweezer method.But this method can only be used for handling, and can't different particles be characterized, and moves with identical speed because different types of particle can both be followed the aperture of same speed, on the motion phenomenon, has no difference.This just can't characterize different particles.
Therefore; If a kind of new method can be provided; Make it possible to utilize this manipulate tools of photoelectricity tweezer to realize the sign of biomone, so just can solve the high and flexible poor problem of general dielectric characterization chip cost, also can excavate the new using value of photoelectricity tweezer simultaneously.
Given this; The present invention proposes a kind of method of utilizing the photoelectricity tweezer that biological particle is characterized: utilize the photoelectricity tweezer to the step-out phenomenon in the biomone driving process (being that the too fast particle that causes of light pattern translational speed can't be caught up with); Measure the maximum synchronous speed of biomone in certain exciting signal frequency scope, can realize the dielectric characterization of particle.
Summary of the invention
Technical matters: the purpose of this invention is to provide a kind of method of utilizing the photoelectricity tweezer micro-nano biomone to be carried out dielectric characterization; General biomone dielectric characterization chip manufacturing cost is high to solve, the baroque defective of microelectrode, also can expand the application of photoelectricity tweezer simultaneously greatly.
Technical scheme: the present invention proposes to use particle (promptly being driven in the process of particle movement at light pattern by the step-out phenomenon in the photoelectricity tweezer driving process; The too fast particle that causes of light pattern translational speed can't be caught up with); Measure biomone in certain exciting signal frequency scope maximum synchronous speed, can realize the dielectric characterization of particle.
At present mostly the dielectric characterization of biomone is to compose through imaginary part that the Clausius-Mosso of measure moving particle is put forward (Clausius-Mossotti) complex factor and realizes that this mainly is in order to utilize electrorotation chip.And in fact, under considerable situation, measure Clausius-Mosso to put forward the real part spectrum of complex factor and also can reach particle is characterized, and then reach the purpose of differentiating and distinguishing.The present invention just is being based on this, utilizes the correlativity of the real part that driving force that the photoelectricity tweezer produced and Clausius-Mosso put forward complex factor, carries the real part of complex factor through the Clausius-Mosso of measure moving particle and composes the dielectric characterization of realizing biomone.
The step-out phenomenon of micro-nano biomone under photoelectricity tweezer manipulation effect will definitely be avoided in little manipulation process, but exactly the feasible dielectric characterization to particle of this phenomenon becomes possibility.Specifically, when particle to be measured during by photoelectricity tweezer driven, particle will be followed by the track of light pattern; But after the translational speed of light pattern increases to a certain degree, particle will not catch up with the motion of light pattern.In other words, there is a maximum synchronous speed in particle when following the light pattern motion.The suffered dielectrophoretic force of particle that is in maximum synchronous speed is maximum; Again because dielectrophoretic force is proportional to the gradient of electric field intensity square; So it is maximum to be in the gradient of electric field intensity square of the residing position of particle of maximum synchronous speed; In other words, particle will reach maximum synchronous speed in the maximum position of the gradient of electric field intensity square.In addition; The maximum synchronous speed of particle and the Clausius-Mossotti factor of particle or effective polarizability (dielectric property that has reflected particle) are closely related, and the frequency dependence of the Clausius-Mossotti factor of particle has determined the maximum synchronous speed of particle to have frequencydependence characteristic.Therefore; The frequency characteristic of the maximum synchronous speed through measure moving particle; And combine the quantitative relationship between the Clausius-Mossotti factor three of maximal value and particle of gradient of maximum synchronous speed, electric field intensity square of particle; Can draw the frequency characteristic of the Clausius-Mossotti factor of particle, and then can parse the dielectric parameter of particle, realize the dielectric characterization of biomone.This new method does not relate to the making of any physical entity microelectrode, and has made full use of the flexibility advantage of photoelectricity tweezer, and therefore this method not only greatly reduces the cost of biomone dielectric characterization, and performance is very superior.
Specifically utilize the photoelectricity tweezer to be: when utilizing light pattern to drive micro-nano biomone to the method that micro-nano biomone carries out dielectric characterization; Make particle reach the maximal rate synchronous through the translational speed that progressively changes light pattern with light pattern; Be that particle is about to not catch up with the critical velocity that light pattern moves; And the frequency adjustment of combination pumping signal, measure the maximum synchronous speed curve of biomone in the certain frequency scope of pumping signal, and then accomplish the dielectric characterization of particle.
The concrete steps of this dielectric characterization method are:
Step 1: the sample solution that will contain intended particle is expelled in each miniflow body cavity of dielectric characterization chip through each injection port;
Step 2: projection dummy electrodes pattern in each the miniflow body cavity on chip;
Step 3: the voltage of sinusoidal excitation signal is added between the upper strata conducting film and lower floor's conductive layer of the chip that uses, makes electric field pass the miniflow body cavity, and set original frequency;
Step 4: in each the miniflow body cavity on chip; Constantly change the translational speed of dummy electrodes pattern; Particle is being pushed away by the dummy electrodes pattern or, reach maximum synchronous speed, and note the maximum synchronous speed in the particle movement process up to particle in tow along straightaway;
Step 5: change the frequency of pumping signal, make frequency hopping to a new Frequency point of pumping signal, repeating step 4 then;
Step 6: draw out the variation spectral line of the maximum synchronous speed of intended particle in the survey frequency scope with exciting signal frequency.
Shifting gears of the translational speed of described dummy electrodes pattern is to start from scratch progressively to increase, or dichotomy changes speed; Even particle is out-of-step free under initial velocity; The mean value of then choosing initial velocity value and most probable velocity value is as next test speed value; Otherwise if particle step-out under initial velocity, then choose mean value between initial velocity value and zero as next test speed value, and the like; Constantly increase or reduce the translational speed of light pattern, until the maximum synchronous speed that finds particle to move with light pattern with dichotomy.
Beneficial effect: the method for utilizing the photoelectricity tweezer micro-nano biomone to be carried out dielectric characterization provided by the invention makes the maximum synchronous speed before particle reaches step-out through the translational speed that progressively increases light pattern; And combine the frequency body of pumping signal to regulate; Measure the maximum synchronous speed of biomone in the certain frequency scope of pumping signal, and then accomplish the dielectric characterization of particle.This method has made full use of the flexibility advantage of photoelectricity tweezer; Avoided on chip, making complicated physical entity electrod-array simultaneously; And particle handled and test is integrated in one, and only need one road sinusoidal excitation signal (present electric rotary test chip needs the sinusoidal signal more than 3 tunnel).Therefore; Dielectric characterization method provided by the invention all is superior to the dielectric characterization method of current micro-and nano-particles at cost, function, aspect of performance; For the great-leap-forward development of biomedical detection range provides very necessary means, will be widely used in fields such as medical diagnosis on disease and treatment, molecular biology, public health quarantine, judicial expertise, Food Hygiene Surveillances.
Description of drawings
Fig. 1 is the employed dielectric characterization chip structure of an embodiment of the invention synoptic diagram;
Fig. 2 is the light pattern that utilizes in the embodiment of the invention photoelectricity tweezer that biomone the is carried out dielectric characterization synoptic diagram (chip is thrown off the vertical view behind the upper substrate) of arranging.
Have among the above figure:
Embodiment
The embodiment that utilizes the photoelectricity tweezer carries out dielectric characterization to biomone method provided by the invention is referring to Fig. 1 and Fig. 2.This method the pattern of chip structure, material and the light pattern that can use be not limited to present embodiment.
Employed dielectric characterization chip comprises first injection port 1101, second injection port 1102, the 3rd injection port 1103 in the present embodiment; Transparent insulation cover plate 120; The transparent indium and tin oxide film in upper strata 130, the first miniflow body cavitys 1401, the second miniflow body cavity 1402, the 3rd miniflow body cavity 1403, middle ware interlayer 150; Transparent insulation substrate 160, the dummy electrodes cambium layer of forming by silicon nitride layer 171, photoconductive layer 172 and transparency conducting layer 173 170.Silicon nitride layer 171 can prevent hydrolysis; Photoconductive layer 172 has photoconductive characteristic; Promptly when by illumination when bright its inner charge carrier quantity increase severely, and not by illumination when bright its inner charge carrier quantity seldom, the ratio that its plain telegram is led with dark conductance can reach more than 10000 at least; The material of photoconductive layer 172 can be selected amorphous silicon hydride or the cadmium sulfide (CdS) that mixes or the cadmium selenide (CdSe) that mixes or the combination of cadmium sulfide and cadmium selenide.The purpose of three miniflow body cavitys is arranged is simultaneously three kinds of different samples particles to be characterized to employed chip in the present embodiment, and three miniflow body cavitys separate three kinds of samples to prevent mutual pollution.In addition, can respectively in three miniflow body cavitys the form characteristics according to corresponding particle throw difform light pattern, be beneficial to the dielectric characterization of various different particles.
In the present embodiment, it is following to utilize the photoelectricity tweezer biomone to be carried out the concrete steps of method of dielectric characterization:
Step 1: be expelled to the sample solution of three kinds of intended particles in the first miniflow body cavity 1401, the second miniflow body cavity 1402, the 3rd miniflow body cavity 1403 of dielectric characterization chip respectively through three injection ports;
Step 2: as shown in Figure 2, projection rectangle dummy electrodes 1801 and circular dummy electrodes 1802 in miniflow body cavity 1401; Projection aperture dummy electrodes 190 in miniflow body cavity 1402; Projection rectangle frame dummy electrodes 200 in miniflow body cavity 1403;
Step 3: the voltage of sinusoidal excitation signal is added between upper strata indium and tin oxide film 130 and the transparency conducting layer 173, and sets original frequency;
Step 4: in miniflow body cavity 1401; Make rectangle dummy electrodes 1801 fixed; Move circular dummy electrodes 1802 according to the direction of arrow among Fig. 2; And start from scratch and increase translational speed gradually, the particle between two rectangle dummy electrodes 1801 is being pushed away by circular dummy electrodes 1802 or in tow along straightaway, is noting the maximum synchronous speed in the particle movement process; In miniflow body cavity 1402; Move aperture dummy electrodes 190 according to the direction of arrow among Fig. 2; And start from scratch and increase translational speed gradually, the particle that is positioned at aperture is by 190 tractions of aperture dummy electrodes and move, and notes the maximum synchronous speed in the particle movement process; In miniflow body cavity 1403; Move rectangle frame dummy electrodes 200 according to the direction of arrow among Fig. 2; And start from scratch and increase translational speed gradually, the particle that is positioned at rectangle frame is by 200 tractions of rectangle frame dummy electrodes and move, and notes the maximum synchronous speed in the particle movement process;
Step 5: change the frequency of pumping signal, make frequency hopping to a new Frequency point of pumping signal, repeating step 4 then;
Step 6: draw out the variation spectral line of the maximum synchronous speed of target organism particle in the survey frequency scope with exciting signal frequency.
Claims (3)
1. method of utilizing the photoelectricity tweezer micro-nano biomone to be carried out dielectric characterization; It is characterized in that: when utilizing light pattern to drive micro-nano biomone; Make particle reach the maximal rate synchronous through the translational speed that progressively changes light pattern with light pattern; Said maximal rate particle is about to not catch up with the critical velocity that light pattern moves; And the frequency adjustment of combination pumping signal, measure the maximum synchronous speed curve of biomone in the certain frequency scope of pumping signal, and then accomplish the dielectric characterization of particle.
2. the photoelectricity tweezer that utilizes as claimed in claim 1 carries out the method for dielectric characterization to micro-nano biomone, it is characterized in that the concrete steps of this dielectric characterization method are:
Step 1: the sample solution that will contain intended particle is expelled in each miniflow body cavity (140) of dielectric characterization chip through each injection port (110);
Step 2: projection dummy electrodes pattern (180) in each the miniflow body cavity (140) on chip;
Step 3: the voltage of sinusoidal excitation signal is added between the upper strata conducting film (130) and lower floor's conductive layer (173) of the chip that uses, makes electric field pass miniflow body cavity (140), and set original frequency;
Step 4: in each the miniflow body cavity (140) on chip; Constantly change the translational speed of dummy electrodes pattern (180); Particle is being pushed away or in tow along straightaway by dummy electrodes pattern (180); Reach maximum synchronous speed up to particle, and note the maximum synchronous speed in the particle movement process;
Step 5: change the frequency of pumping signal, make frequency hopping to a new Frequency point of pumping signal, repeating step 4 then;
Step 6: draw out the variation spectral line of the maximum synchronous speed of intended particle in the survey frequency scope with exciting signal frequency.
3. the photoelectricity tweezer that utilizes as claimed in claim 2 carries out the method for dielectric characterization to micro-nano biomone, it is characterized in that, shifting gears of the translational speed of described dummy electrodes pattern (180) is to start from scratch progressively to increase, or dichotomy changes speed; Said dichotomy refers to if particle is out-of-step free under initial velocity; The mean value of then choosing initial velocity value and most probable velocity value is as next test speed value; Otherwise if particle step-out under initial velocity, then choose mean value between initial velocity value and zero as next test speed value, and the like; Constantly increase or reduce the translational speed of light pattern, until the maximum synchronous speed that finds particle to move with light pattern with dichotomy.
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| CN102384980B (en) * | 2010-08-30 | 2014-03-26 | 明达医学科技股份有限公司 | Micro-fluid control device and operation method thereof |
| DE102013205540A1 (en) * | 2013-03-28 | 2014-10-02 | Robert Bosch Gmbh | Sensor element and method for detecting a gas |
| CN105044192B (en) * | 2015-08-14 | 2018-11-02 | 深圳大学 | A kind of cell sorting method based on light-induction dielectrophoresis technology |
| US10495580B2 (en) * | 2015-10-07 | 2019-12-03 | Afi Corporation | Inspection device, inspection system, and inspection method |
| CN105602836B (en) * | 2016-02-03 | 2017-10-27 | 河海大学常州校区 | Photo-dielectric swimming assembling chip and the method for building structure gradient Gel Compositions body |
| CN109679845B (en) * | 2019-01-16 | 2020-09-22 | 江南大学 | Microbial cell factory constructed based on light-induced dielectrophoresis technology and application thereof |
| CN113312829B (en) * | 2021-05-26 | 2022-07-26 | 江南大学 | Micro-nano particle movement control method based on data-driven ODEP kinematic model |
| CN116899644B (en) * | 2023-09-12 | 2023-11-28 | 微纳动力(北京)科技有限责任公司 | Photoelectric micro-fluidic device and system |
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