CN104535479B - For the Asia-Pacific hertz nano biological sensor of single or a small amount of cell detection - Google Patents
For the Asia-Pacific hertz nano biological sensor of single or a small amount of cell detection Download PDFInfo
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Abstract
The invention discloses the Asia-Pacific hertz nano biological sensor for single or a small amount of cell detection, belong to sensor technical field, including the Terahertz pattern detection pond that microfluidic elements and slit nano-antenna are constituted, the microfluidic elements include at least one miniflow body in tubular construction, are arranged on the malleation generating means at miniflow body cell entry end, are arranged on the depression generator at miniflow body cell outlet end, the slit nano-antenna is provided through the elongate slot of antenna body, and the miniflow body is corresponded through elongate slot.By setting the Terahertz pattern detection pond that microfluidic elements and slit nano-antenna are constituted, cell to be measured can be allowed be single or a small number of to be amplified treatment by and to the signal of individual cells, it is real to realize with the direct qualitative and quantitative determination various trait bacterium of Asia-Pacific hertz sensing technology by capturing unicellular signal and then the cell of various trait can be made a distinction and be recognized.
Description
Technical field
The invention belongs to sensor technical field, and in particular to a kind of Asia-Pacific for single or a small amount of cell detection hertz
Nano biological sensor.
Background technology
At present, Pasteur and kock are in the master that the Bacteria Culture set up in 19th century and identification are still clinical microbiology laboratory technique
Flow Technique and method.Because the Bacteria Culture cycle is long, bacterium Species estimation and drug sensitive test generally require to take one week or so.This
Outward, also there is culture matrix difference, incubation time long (such as：Tubercle bacillus needs the incubation time of nearly 1 month), be difficult to culture and
Separate (such as：Haemophilus influenzae) etc. problems.Over nearly 20 years, in order to overcome above-mentioned drawback, occur in that Series Molecules are biological
Technology and method is learned, mainly including real-time fluorescence PCR, digital pcr, genetic chip, biology sensor, bacterium mass spectrum, the second generation
Sequencing technologies etc., portion of techniques or method are in routine clinical development.Although portion of techniques and method are (such as：Real-time fluorescence
PCR Bacteria Culture) can be avoided, but can only often detects several limited common pathogens, and cannot be distinguished by dead bacterium and work
Bacterium.Although MALDI _ TOFMS is known as " revolutionary microbial identification in minutes ", should
The detection sensitivity of technology is low (i.e.：104~105CFU), it is still its essential prior step that Bacteria Culture obtains pure bacterium,
Meanwhile, apparatus expensive, maintenance cost is high, it is difficult to differentiate some bacterium kinds and variant (such as：Escherichia coli and Shigella).
It can be seen that, " Bacteria Culture of forgoing link, directly qualitative and quantitative determination various trait bacterium (kind, growth conditions, drug resistance) "
It is key scientific problems that the quick detection of bacterium must be solved.This cannot be almost solved due to existing Protocols in Molecular Biology to ask
Topic, therefore, it is necessary to seek to can effectively solve the problem that the interdisciplinary detection theory and technique of above-mentioned technical bottleneck.
Terahertz is that interaction weak between electromagnetic wave of the frequency in 0.1~10THz, large biological molecule includes hydrogen bond and model
The low-frequency vibrations such as De Huali, skeletal vibration and dipole rotation absorb and are respectively positioned in Terahertz frequency range, when THz wave is by life
During thing macromolecular, its low frequency modes is excited, cause energy level transition, according to absorption peak position different on absorption spectrum and by force
" the THz fingerprint characteristics " of degree realizes the sign of material.Early stage Terahertz detection technique mainly applies to solid matter, by peculiar
Absworption peak and absorption coefficient carry out the single identification of material.This presence for being mainly a large amount of hydrogen bonds in water has big to THz wave
Amount absorbs.During large biological molecule in THz wave direct measurement liquid phase sample, due to a large amount of absorptions of hydrone, letter is can detect
It is number weaker and repeated poor seriously limit the utilization of its medical domain.The water molecule layer on solute surface, water particularly in the aqueous solution
Molecule in close is combined, complex structure, and hydrogen bond largely exists to produce the terahertz absorption spectra of solute to be influenceed strongly, severe jamming
The sign of sample in solution.In recent years, the THz wave of sub- terahertz wave band, i.e. frequency less than 1THz, causes all the more people
Interest.Globus et al. has found that water ambient interferences suffered by liquid phase detection material are very small in this wave band, can understand and manifest
Various resonance in solution in large biological molecule caused by low frequency modes.In the wavelength band of Asia-Pacific hertz, hydrone
Absorb and at least reduce 2.5 orders of magnitude compared with infrared and far infrared band so that test substance spectral signature in itself is obtained
To express.In the range of Asia-Pacific hertz, they propose 10cm again-1To 25cm-1Frequency field bacterium sensitive can must be shown not
With the difference of spectral signature under environment, can the difference that must be verified between variety classes thalline sensitive enough, this be subsequent experimental light
The selection of frequency range provides important references during spectrum interpretation of result.Nearly more than 2 years research with Escherichia coli as model organism
Result shows that the Asia-Pacific hertz sensing technology for overcoming water sensitivity can be by the Asia-Pacific of the whole thalline of bacterium hertz vibration light
Spectrum directly distinguishes Escherichia coli and bacillus subtilis.In order to verify the sign ability of sub- Terahertz Technology, this laboratory uses
Effective frequency is 0.2-3 Terahertzs, and the terahertz time-domain spectroscopy instrument of wavelength 1000-1500um, detection sample concentration is 108CFU/
The EHEC of ml and the bacterium solution of staphylococcus aureus, detection means are that internal diameter is the quartz colorimetric utensil of 100um, are as a result shown
Show that both absorption spectrums show notable difference, fully confirm that Asia-Pacific hertz sensing technology is used for the reason of bacterium direct detection
By with technical feasibility.Asia-Pacific hertz sensing technology efficiently solves " water sensitivity " problem, possesses direct qualitative and quantitative inspection
Survey the physical features and unique advantage of various trait bacterium.
Although existing Asia-Pacific hertz sensing technology possesses direct detection bacterium ability, detection is reached single celled sensitive
Degree, it is necessary to solve two key issues：Individual cells are by terahertz light source detection；Possesses recognizable single celled noise
Than.
For problem one：The scheme of solution can be that the caliber of microfluid is set into or bigger one identical with cell size
A bit, it can be ensured that can only cell by microfluid, but because microfluid caliber is too small, increase of the inside pipe wall to cell resistance makes
Cell can not be normal through causing detection to carry out, while cannot also be processed manufacture；
For problem two：Can solve the problems, such as to recognize unicellular signal to noise ratio with the method for improving detection sensitivity, but
Also more highly sensitive detecting instrument is not matched with microfluid conventional process at this stage.
Thus, it is necessary to research and development one kind can make that single or a few cell passes through microfluid and possess can regarding to the issue above
The Asia-Pacific hertz nano biological sensor of unicellular spectral signal ability is recognized, and researches and develops the detection method of the sensor, make inspection
Survey cell be it is single be present in microfluid by microfluid and with the quantity for determining, thoroughly solve the problems, such as Single cell analysis.
The present invention intends on original sub- THz sensing technologies basis, and plan is respectively adopted microfluid and slit after improvement
Nano-antenna technology solves above-mentioned two key technical problem, and will incorporate the Asia of microfluid and slit nano-antenna technology
THz sensing technologies are referred to as sub- THz nano biological sensors.Microfluid refers to control, operate and detect multiple under microscopic dimensions
Microfluidic channel diameter is narrowed down to 5um or so by the technology of miscellaneous fluid, the present invention from traditional 50um, and in this body cell of miniflow
Malleation generating means and depression generator that arrival end and the port of export are respectively provided with.Microfluid sample preparation pond master after improvement
If for separating and prepare unicellular bacterium solution, making that the cell of bacterium to be checked at THz light source detections is single must to be solved by detection cell
The certainly specific problem of detection architecture.Slit nano-antenna is the nanometer slot configurations of metal nanoparticle composition, possesses spectrum
The extreme amplifying power of signal.After the nano-metal particle of slit nano-antenna is subject to sub- THz radiation, its is distinctive asymmetric
Electromagnetic field environment can greatly strengthen the local surface plasma resonance of nanometer slit, inhale medium molecule spectrum in slit
Receiving cross section increases by 103~105More than times, absorption coefficient also extremely increases to 106~107cm-1, finally make the inspection of medium molecule
Survey sensitivity and improve 103~1012Times.The existing sensitivity (10 of sub- THz sensing technologies detection bacterium2~103CFU) it is unable to reach
Single cell analysis level, can make detection spectral signal amplify more than 1000 times using slit nano-antenna.By slit nano-antenna
It is incorporated into microfluid microcavity, it becomes possible to sub- THz nano biological sensors is possessed the spirit of direct detection bacterium individual cells
Sensitivity, realizes bacterium direct detection.Relative to existing bacteriological labororatory's detection technique, the Asia-Pacific of Single cell analysis is really realized
Hertz nano biological sensor is " revolutionary microorganism detection new technology and new method in minutes ", with being trained without bacterium
Support, without mark, without reagent, real-time detection, easy quick (2~3 minutes), the directly qualitative and quantitative inspection of mixed infection sample
Many advantages, such as survey.
The content of the invention
In view of this, it is an object of the invention to provide a kind of Asia-Pacific for single or a small amount of cell detection hertz nanometer
Biology sensor, to solve cannot to realize in the prior art single or a small amount of cell by that at terahertz light source detection and cannot know
The problem of not single celled signal to noise ratio, there is provided following technical scheme：
For the Asia-Pacific hertz nano biological sensor of single or a small amount of cell detection, including Asia-Pacific hertz sensing technology,
Microfluidic elements and slit nano-antenna, the microfluidic elements include at least one miniflow body, setting in tubular construction
Malleation generating means at miniflow body cell entry end, the depression generator for being arranged on miniflow body cell outlet end, institute
The elongate slot that slit nano-antenna is provided through antenna body is stated, the miniflow body is corresponded through elongate slot.
Further, the miniflow body is to be arranged in parallel or the setting of Y types or U-shaped setting.
Further, the miniflow body bore is 4-6 μm, and the groove width of the elongate slot is matched with microfluid bore and set
Put.
Further, the malleation generating means includes pressure chamber I, solution cavity I, is set between the pressure chamber I, solution cavity I
Be equipped with isolation push pedal I, be provided with inlet and outlet I in the pressure chamber, the solution cavity I be provided with an inlet I and with it is micro-
The connection of stream body and quantity identical liquid outlet I.
Further, it is provided with one-way control valve on the inlet I.
Further, the malleation generating means is micropump.
Further, the depression generator include pressure chamber II, solution cavity II, the pressure chamber II, solution cavity II it
Between be provided with pressure chamber II described in isolation push pedal II and be provided with inlet and outlet II, the solution cavity II be provided with liquid outlet II and
Connected with miniflow body and quantity identical inlet II.
Further, it is provided with one-way control valve on the liquid outlet II.
Further, the depression generator is peristaltic pump.
The beneficial effects of the present invention are：The present invention is utilized at microfluid body two ends by setting microfluidic elements
The malleation of setting, depression generator, can produce enough negative pressure in miniflow body, allow the cell to be measured can only single or minority
Pass through, so as to effectively improve the specificity of detection, in addition, by the slit nano-antenna that is arranged on miniflow body to unicellular
Signal extremely amplified, effectively increase the sensitivity of detection, the cell of various trait is made a distinction and recognized, really
Realize with Asia-Pacific hertz sensing technology is directly qualitative and quantitative determination various trait bacterium, be conducive to forgoing Bacteria Culture link and
Type specimen liquid measure is reduced, so as to improve detection efficiency and quality.
Other advantages of the invention, target and feature will be illustrated in the following description to a certain extent, and
And to a certain extent, based on being will be apparent to those skilled in the art to investigating hereafter, Huo Zheke
To be instructed from the practice of the present invention.Target of the invention and other advantages can be realized by following specification and
Obtain.
Brief description of the drawings
In order that the purpose of the present invention, technical scheme and beneficial effect are clearer, the present invention provides drawings described below：
Fig. 1 is the structural representation of first embodiment of the invention；
Fig. 2 is the structural representation of first embodiment of the invention malleation generating means；
Fig. 3 is the structural representation of first embodiment of the invention depression generator；
Fig. 4 is the structural representation of second embodiment of the invention；
Fig. 5 is the structural representation of third embodiment of the invention.
Specific embodiment
Hereinafter with reference to accompanying drawing, the preferred embodiments of the present invention are described in detail.It should be appreciated that preferred embodiment
Only for the explanation present invention, rather than in order to limit the scope of the invention.
Embodiment 1
As shown in Figure 1 to Figure 3, the present embodiment includes microfluidic elements, slit nano-antenna 4, the microfluidic elements bag
At least one miniflow body 1 in tubular construction is included, the malleation generating means 2 at the cell entry end of miniflow body 1 is arranged on, is set
In the depression generator 3 at miniflow body cell outlet end, malleation generating means produces positive pressure, and depression generator is produced
Negative pressure, under positive/negative pressure collective effect, by the cell propulsion miniflow body 1 of cell solution, realizes detection, and the slit is received
Rice antenna 4 is provided through the elongate slot 5 of antenna body, and the miniflow body 1 is corresponded through elongate slot 5, when to be measured thin
Born of the same parents from microfluid pass through elongate slot when by signal when row amplify.
The present embodiment occurs by setting microfluidic elements using the malleation, negative pressure that are set at microfluid body two ends
Device, can produce enough negative pressure in miniflow body, allow cell to be measured can only single or minority pass through, so as to effectively improve inspection
The specificity of survey, in addition, single celled signal is amplified by the slit nano-antenna being arranged on miniflow body, will not
Make a distinction and recognize with the cell of proterties, it is real to realize with the direct qualitative and quantitative determination dissimilarity of Asia-Pacific hertz sensing technology
Shape bacterium, is conducive to forgoing Bacteria Culture link and reduces type specimen liquid measure, so as to improve detection efficiency and quality.
In the present embodiment, the miniflow body bore is 4-6 μm, preferably 5 μm, the groove width and miniflow of the elongate slot
The matching of body bore is set, convenient to insert in elongate slot miniflow body, increases malleation using at microfluid body two ends, bears
Pressure generating apparatus, can produce enough negative pressure in miniflow body, allow cell to be measured can only single or minority pass through so that effectively
The sensitivity of detection is improved, is met with directly qualitative and quantitative determination various trait bacterium the specificity of Asia-Pacific hertz sensing technology
It is required that.
The miniflow body of the present embodiment is configured by the way of being parallel to each other, so can be to single miniflow sheet
Body is produced, and processing is easier, and survival rate is high, advantageously reduces manufacturing cost.
In the present embodiment, the miniflow body diameter is more bigger than cell size on the premise of it can fabricate,
But caliber is smaller, and it is bigger to cell resistance, causes cell to be easily detected by, a diameter of 5 μm of the preferred miniflow body of the present embodiment,
The effect for being further added by positive/negative-pressure generating means with this understanding is best.
In the present embodiment, the malleation generating means can be micropump, motor etc., and the preferred malleation of the present embodiment occurs
Device includes pressure chamber I 201, solution cavity I 202, and isolation push pedal is provided between the pressure chamber I 201, solution cavity I 202
I 203, is provided with inlet and outlet I 204 in the pressure chamber, the solution cavity is provided with an inlet I 205 and and miniflow
Body connect and with microfluid quantity identical liquid outlet I 206, be provided with one-way control valve I on the inlet I205
207.After enough cell solutions are loaded in solution cavity I 201, inlet and outlet I 204 enters compressed air, promotes isolation
Push pedal I203 reduces the volume of solution cavity I202, so as to increase the hydraulic coupling of its cell solution, makes it into miniflow body, together
When, setting one-way control valve I207 can prevent solution from flowing backwards the interior pressure for ensureing solution cavity I2023.In the present embodiment, also may be used
It is micropump
In the present embodiment, the malleation generating means can be peristaltic pump, motor etc., and the preferred negative pressure of the present embodiment occurs
Device includes pressure chamber II 301, solution cavity II 302, and isolation is provided between the pressure chamber II 301, solution cavity II 302
Be provided with inlet and outlet II 304 in push pedal II 303, the pressure chamber II 301, the solution cavity II 302 be provided with it is micro-
Stream body connection and with microfluid quantity identical inlet II 305 and liquid outlet II 306, set on the liquid outlet II 306
It is equipped with one-way control valve II 307.When that need to detect, when malleation generating means 2 works, depression generator 3 is also opened simultaneously
Dynamic, the discharge compressed airs of inlet and outlet II 304, solution cavity II 302 forms negative pressure, promotes isolation push pedal II 304 to increase solution
The volume of chamber II 302, so that reducing its pressure produces negative pressure, by cell solution suction miniflow body 1, realizes detection, together
When, setting one-way control valve II 307 can prevent solution from flowing backwards the interior pressure for reducing solution cavity II.
In addition, the pressure medium of malleation, depression generator can also be liquid.
Embodiment 2
As shown in figure 4, miniflow body 1 is set to y-type structure by embodiment 2 as different from Example 1, miniflow will be stated
Body merges into unified channel, convenient to focus on the pipeline of multiple miniflow bodies 1, shape before the connection of depression generator 3
Into negative pressure structure, cell solution is more easy to enter miniflow body 1, better.Other structures are same as Example 1, different here
Ao Shu.
Embodiment 3
If Fig. 5 is embodiment 3, miniflow body 1 is set to U-shape structure as different from Example 1, this structure can be with
Increase the length of miniflow body, it is easier to disperse the cell to be checked of quantification, be conducive to cell solution to enter miniflow body
1, it is better.Other structures are same as Example 1, here no longer Ao Shu.
Finally illustrate, preferred embodiment above is merely illustrative of the technical solution of the present invention and unrestricted, although logical
Cross above preferred embodiment to be described in detail the present invention, it is to be understood by those skilled in the art that can be
Various changes are made to it in form and in details, without departing from claims of the present invention limited range.
Claims (8)
1. the Asia-Pacific hertz nano biological sensor of single or a small amount of cell detection, including microfluidic elements are used for, and its feature exists
In：Also include slit nano-antenna, the microfluidic elements include at least one miniflow body in tubular construction, be arranged on it is micro-
The malleation generating means at stream body cell entry end, the depression generator for being arranged on miniflow body cell outlet end, it is described narrow
Groove nano-antenna is provided through the elongate slot of antenna body, and the miniflow body is corresponded through elongate slot, the miniflow
Body bore is 4-6 μm, and the groove width of the elongate slot matches setting with microfluid bore.
2. the Asia-Pacific for single or a small amount of cell detection according to claim 1 hertz nano biological sensor, it is special
Levy and be：The miniflow body is to be arranged in parallel or the setting of Y types or U-shaped setting.
3. the Asia-Pacific for single or a small amount of cell detection according to claim 1 hertz nano biological sensor, it is special
Levy and be：The malleation generating means includes pressure chamber I, solution cavity I, and isolation is provided between the pressure chamber I, solution cavity I
Push pedal I, is provided with inlet and outlet I in the pressure chamber, the solution cavity is provided with an inlet I and is connected with miniflow body
And quantity identical liquid outlet I.
4. the Asia-Pacific for single or a small amount of cell detection according to claim 3 hertz nano biological sensor, it is special
Levy and be：One-way control valve is provided with the inlet I.
5. the Asia-Pacific for single or a small amount of cell detection according to claim 1 hertz nano biological sensor, it is special
Levy and be：The malleation generating means is micropump.
6. the Asia-Pacific for single or a small amount of cell detection according to claim 1 hertz nano biological sensor, it is special
Levy and be：The depression generator includes pressure chamber II, solution cavity II, is provided between the pressure chamber II, solution cavity II
Isolation push pedal II, is provided with inlet and outlet II, the solution cavity II and is provided with liquid outlet II and and miniflow in the pressure chamber II
Body is connected and quantity identical inlet II.
7. the Asia-Pacific for single or a small amount of cell detection according to claim 6 hertz nano biological sensor, it is special
Levy and be：One-way control valve is provided with the liquid outlet II.
8. the Asia-Pacific for single or a small amount of cell detection according to claim 1 hertz nano biological sensor, it is special
Levy and be：The depression generator is peristaltic pump.
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105505759B (en) * | 2016-01-28 | 2017-09-29 | 中国人民解放军第三军医大学第一附属医院 | The rolling circle amplification Terahertz Meta Materials biology sensor and its detection method of a variety of pathogen Parallel testings |
| CN106483059B (en) * | 2016-10-13 | 2019-03-19 | 中国人民解放军第三军医大学第一附属医院 | Terahertz fluidic cell sensor and its detection method for single or a small amount of living cells markless detection |
| CN107446807B (en) * | 2017-07-26 | 2019-08-06 | 中国人民解放军陆军军医大学第一附属医院 | Integrated form Terahertz superstructure nano biological chip and its application and method |
| CN110132881B (en) * | 2019-02-02 | 2020-01-21 | 中国人民解放军陆军军医大学第一附属医院 | Multispectral nano array chip integrating terahertz spectrum and Raman spectrum and application |
| CN110308107B (en) * | 2019-07-01 | 2021-09-28 | 中国人民解放军陆军军医大学第一附属医院 | Terahertz metamaterial device for detecting cell activity and using method |
| CN111551514A (en) * | 2020-05-08 | 2020-08-18 | 南京大学 | High-sensitivity terahertz sensor capable of detecting trace cells and detection method |
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