CN208171807U - A kind of lymphocyte Photobiology sensor based on optofluidic capillary microcavity - Google Patents
A kind of lymphocyte Photobiology sensor based on optofluidic capillary microcavity Download PDFInfo
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- CN208171807U CN208171807U CN201820800670.5U CN201820800670U CN208171807U CN 208171807 U CN208171807 U CN 208171807U CN 201820800670 U CN201820800670 U CN 201820800670U CN 208171807 U CN208171807 U CN 208171807U
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Abstract
The utility model discloses a kind of lymphocyte Photobiology sensor based on optofluidic capillary microcavity, the sensor includes frequency swept laser, for carrying out capillary microcavity-micro-nano fiber coupling unit of sensing testing to cell solution, photodetector and feedback control system, frequency swept laser, capillary microcavity-micro-nano fiber coupling unit, photodetector and feedback control system are of coupled connections by fiber fuse between each other, feedback control system is electrically connected with photodetector and frequency swept laser respectively, capillary microcavity-micro-nano fiber coupling unit is mutually perpendicular to be coupled to form by micro-nano fiber and capillary microcavity.The sensor uses micro volume, heightQThe capillary microcavity of value is as lymphocyte sensing unit, and by the control and interaction of light in microcavity and microfluid, realizing has highly sensitive, quickly detection, the Photobiology lymphocyte sensing that unmarked, compact-sized, integrated level is high, at low cost.
Description
Technical field
The utility model relates to a kind of lymphocyte Photobiology sensors based on optofluidic capillary microcavity, can be used for
Sensor technical field.
Background technique
It was found that and quantization AIDS virus ability for doctor seek maximum therapeutic effect be it is vital,
And realize that a kind of method of this target is exactly to observe the quantity of CD4+ lymphocyte.The CD4+ cell of one healthy normal person
In 1200/ μ L, with the intensification of AIDS gradient of infection, this number can simultaneously decline number, be examined when dropping to 200/ μ L
Break as AIDS.
Currently, the counting of CD4+ lymphocyte can be accurately realized by flow cytometer.The work of these instruments
Principle is by marking CD4+ receptor with fluorescence antibody and counting to the fluorescence signal therefore generated.But while streaming is thin
The counting of born of the same parents' instrument is very accurate, they also have dependent on fluorescent marker, equipment cost is high, maintenance requirement is high, detection time is long
Etc. limitations.
Summary of the invention
The purpose of this utility model is exactly to propose a kind of based on light stream to solve the above-mentioned problems in the prior art
The lymphocyte Photobiology sensor of hair control tubule microcavity.
The purpose of this utility model will be realized through the following technical scheme：It is a kind of based on optofluidic capillary microcavity
Lymphocyte Photobiology sensor, including frequency swept laser, the capillary microcavity-for carrying out sensing testing to cell solution
Micro-nano fiber coupling unit, photodetector and feedback control system, the frequency swept laser, capillary microcavity-micro-nano fiber
Coupling unit, photodetector and feedback control system are of coupled connections by fiber fuse between each other, the feedback control system
System is electrically connected with photodetector and frequency swept laser respectively, and the capillary microcavity-micro-nano fiber coupling unit is by micro-nano
Optical fiber is mutually perpendicular to be coupled to form with capillary microcavity.
Preferably, the wavelength of the frequency swept laser is tunable.
Preferably, the micro-nano fiber is by single mode optical fiber by drawing cone machine fused biconical taper to be made.
Preferably, the capillary microcavity is prepared by hollow quartz capillary optical fiber progress fused biconical taper.
Imitative selection of land, the micro-nano fiber are close proximity on capillary microcavity, the axis of the micro-nano fiber and capillary microcavity
Line keeps antarafacial vertical.
Preferably, the capillary microcavity is high symmetrical hollow cylinder shape structure.
Preferably, only meet Whispering-gallery-mode resonance condition resonance wavelength can be generated in capillary microcavity it is humorous
Vibration, the resonance wavelength for meeting Whispering-gallery-mode resonance condition are determined by following formula：
The π of λ=2 rneff/m
Wherein r is resonant cavity radius, neffIt is the effective refractive index that resonant optical mode passes through, m is integer.
Preferably, resonant cycle period of the light wave in capillary microcavity determines have by the quality factor q of capillary microcavity
Imitate action length LeffIt is given by with quality factor q relationship：
Leff=Q the π of λ/2 neff。
The advantages of technical solutions of the utility model, is mainly reflected in：The sensor is using micro volume, the capillary of high q-factor
Microcavity is as lymphocyte sensing unit, by the control and interaction of light in microcavity and microfluid, realize have it is highly sensitive,
Quickly detection, the Photobiology lymphocyte sensing that unmarked, compact-sized, integrated level is high, at low cost.By to all -fiber coupling
The mode of conjunction excites and the Whispering-gallery-mode resonance spectrum in detection optical microcavity, under the means without any fluorescent marker, knot
Light combination flow control/microflow control technique realizes the detection to cell concentration, has all -fiber, quick, unmarked, inexpensive, repeatable
The characteristics of test.The Photobiology sensor that the utility model is realized has in the pre- diagnosing and treating for solving the problems, such as AIDS
There is potential, huge application value.
Detailed description of the invention
Fig. 1 is the utility model optical sensor system structural schematic diagram.
Fig. 2 is that the utility model optical sensor cross-sectional view and light field transmit schematic diagram.
Fig. 3 is the non-functionalization of the utility model microcavity inner wall surface, each leads into water, phosphate-buffered salt, solution, 1XCD4+
The Whispering-gallery-mode resonance spectrum tested after T cell liquid, 10XCD4+T cell PBS solution.
Fig. 4 is that echo wall die resonance Frequency bias changes over time situation during the utility model is antibody functionalized.
Fig. 5 is that the utility model microcavity inner wall surface is implanted into after CD4+ antibody, is passed through into microcavity thin containing 1XCD4+T
The Whispering-gallery-mode resonance spectrum that different time test obtains after the PBS solution of born of the same parents.
Fig. 6 is that the wave length shift of the utility model Whispering-gallery-mode resonance spectrum changes with time situation, wave length shift
About 6.8pm.
Fig. 7 is that the utility model microcavity inner wall surface is implanted into after CD4+ antibody, is passed through into microcavity containing 10XCD4+T
The Whispering-gallery-mode resonance spectrum that different time test obtains after the PBS solution of cell.
Fig. 8 is that the wave length shift of the utility model Whispering-gallery-mode resonance spectrum changes with time situation, wave length shift
About 16pm.
Specific embodiment
The purpose of this utility model, advantage and feature will carry out figure by the non-limitative illustration of preferred embodiment below
Show and explains.These embodiments are only the prominent examples using technical solutions of the utility model, all to take equivalent replacement or wait
Effect transformation and formed technical solution, all fall within the requires of the utility model protection within the scope of.
The utility model discloses a kind of lymphocyte Photobiology sensor based on optofluidic capillary microcavity, such as schemes
Shown in 1, including frequency swept laser 1, capillary microcavity-micro-nano fiber coupling unit for carrying out sensing testing to cell solution
2, photodetector 3 and feedback control system 4, the frequency swept laser 1, capillary microcavity-micro-nano fiber coupling unit 2, light
Electric explorer 3 and feedback control system 4 are of coupled connections by fiber fuse between each other, the feedback control system 4 respectively with
Photodetector 3 and frequency swept laser 1 are electrically connected, and the wavelength of the frequency swept laser is tunable, the capillary microcavity
For high symmetrical hollow cylinder shape structure, the capillary microcavity is used as cell solution channel in sensing unit, and analyte is logical
Road is separated with detection channels, the influence of hermetic section external world uncertain factor, improves system stability.
Sensing testing, the capillary microcavity-are carried out to cell solution using capillary microcavity-micro-nano fiber coupling unit
Micro-nano fiber coupling unit 2 is mutually perpendicular to be coupled to form by micro-nano fiber 21 and capillary microcavity 22, conical fiber and capillary
The coupling distance of microcavity accurately controls Nano grade, and the capillary microcavity wall thickness used is only several microns, light in microcavity
Nearby by cell solution contact zone field width, interaction efficiency is high for field distribution and microcavity surface, high with the coupling efficiency of conical fiber,
Echo wall die resonance spectrum has larger Q value, it can be achieved that high sensitivity detection.
The frequency swept laser 1 issues laser and enters micro-nano fiber cone area from one end of micro-nano fiber, passes through evanscent field coupling
Close enter capillary microcavity, excite capillary microcavity in Whispering-gallery-mode resonance, Whispering-gallery-mode light field with it is thin inside microcavity
The interaction of specific biological optics occurs for cell lysis liquid, demodulates tested lymph by the light intensity of detection micro-nano fiber other end output
Cell solution cell concentration information, the feedback control system is by being electrically connected the output wavelength of control frequency swept laser and strong
Degree, while also the detection of control photodetector is realized by the luminous intensity of another output end of micro-nano fiber after capillary microcavity
The control of optical wavelength and optical power to frequency swept laser and the light intensity for measuring micro-nano fiber output end and input laser light
Feedback op is compared at specific wavelength of light by force, obtains the Whispering-gallery-mode resonance spectrum with multiple transmission peaks.
For excitation Whispering-gallery-mode resonance, micro-nano fiber and capillary microcavity are vertical coupled, and micro-nano fiber is close proximity to capillary
On pipe microcavity, the two central axes keep antarafacial it is vertical, which can make microcavity after being passed through tested cell sample liquid, still with it is micro-
The couple state that nano fiber keeps relative stability increases intracavitary distribution of light intensity to improve coupling efficiency, and acquisition Q value is higher, transmits
The deeper Whispering-gallery-mode resonance spectrum in peak, guarantees the sensitivity and stability of sensor.
The micro-nano fiber is by single mode optical fiber by drawing cone machine fused biconical taper to be made, and cone area's diameter is 2-3 μm, institute
It states capillary microcavity to be prepared by hollow quartz capillary optical fiber progress fused biconical taper, outer diameter is 90 μm, 2-3 μm of wall thickness, should
Capillary microcavity has the characteristics that high symmetry, thin-walled, volume are small.
Fig. 2 is the cross-sectional view and light field transmission schematic diagram of the technical program lymphocyte Photobiology sensor.
Light field is coupled in capillary thin-walled microcavity in the form of evanscent field from micro-nano fiber cone area in micro-nano fiber cone area, due to total reflection
Effect, the light for meeting the specific wavelength of condition of resonance form Whispering-gallery-mode resonance in Microsphere Cavities, in optical field distribution such as Fig. 2
Shown in 10- Whispering-gallery-mode optical field distribution, most of light energy distribution in close to capillary microcavity internal wall surface regions near,
And there is minimum mode volume, therefore the interaction of light and measured object is very strong, and highly sensitive sensing may be implemented.Work as microcavity
In when being passed through different samples, Whispering-gallery-mode resonance spectrum can because microcavity inner refractive index changes and to the sample of various concentration
Product generate different degrees of Frequency bias.
The resonance wavelength for only meeting Whispering-gallery-mode resonance condition can generate resonance in capillary microcavity, meet
The resonance wavelength of Whispering-gallery-mode resonance condition is determined by following formula：
The π of λ=2 rneff/m
Wherein r is resonant cavity radius, neffIt is the effective refractive index that resonant optical mode passes through, m is integer.
Resonant cycle period of the light wave in capillary microcavity determine by the quality factor q of capillary microcavity, useful effect
Length LeffIt is given by with quality factor q relationship：
Leff=Q the π of λ/2 neff。
The resonance wavelength for only meeting Whispering-gallery-mode resonance condition can generate resonance in capillary microcavity, meet
The resonance wavelength of Whispering-gallery-mode resonance condition is determined by following formula：
The π of λ=2 rneff/m
Wherein r is resonant cavity radius, neffIt is the effective refractive index that resonant optical mode passes through, m is integer.Due to echo
Wall mode resonance light field has longer time photon lifetime, the Photobiology interaction between light field and tested lymphocyte solution
It is greatly increased with length and time.Wherein, interaction length depends on light in the microcavity determined by the quality factor q of microcavity
Wave cycle period determined, effective interaction length LeffIt is given by with quality factor q relationship：
Leff=Q the π of λ/2 neff
By taking the capillary microcavity and laser light source that the technical program uses as an example, quality factor q 106, effective refractive index
neff=1.45, when wavelength X=1.5 μm, effective interaction length is up to 15cm or more.
The lymphocyte Photobiology sensor, is used for biological test for by optical device, has carried out surface to microcavity
Functionalization is to improve specific interaction significance of the microcavity in test lymphocyte solution processes.It is logical first in order to compare
Cross experiment test not to microcavity inner wall carry out it is surface-functionalized, by minute-pressure pump and microflow control technique, be passed through not into microcavity
With the variation characteristic of the Whispering-gallery-mode resonance spectrum of concentrations of cells solution.It is illustrated in figure 3 and each leads into water, phosphate-buffered salt
It is surveyed after (phosphate buffer saline, PBS) solution, 1XCD4+T cell PBS solution, 10XCD4+T cell PBS solution
Obtained Whispering-gallery-mode resonance spectrum is tried, it is measurand and antigen that CD4+T cell, which is one kind of CD4+ lymphocyte, right
It is determining that the antibody of characteristic opposite sex interaction is wanted in certain type of lymphocyte antigen and its progress test process.?
In the technical program, 1X, 10X concentration respectively correspond 1000CD4+T cell/μm and 10000CD4+T cell/μm, similarly hereinafter.It can be with
Find out that PBS solution is led in water flowing again later, spectrum changes, and after having led to PBS solution, in the non-functionalization of inner wall surface
Microcavity in lead to the CD4+T cell liquid of 1X and 10X, spectrum has drifted about 6.8pm and 16pm respectively, this is because certain density thin
Cell lysis liquid can cause variations in refractive index in microcavity, so as to cause the variation of Whispering-gallery-mode resonance spectrum.
Higher sensitivity in order to obtain, we carry out functionalization to microcavity inner wall, and concrete operations are as follows：
S1：It is passed through deionized water inside to capillary microcavity, to cleaning inside capillary microcavity；
S2：It is 5% dust technology to concentration is passed through inside the capillary microcavity after the cleaning of S1 step, and liquid is sealed up for safekeeping
In standing 30 minutes 2 hours in capillary microcavity, after be alternately passed through with deionized water and alcohol, by capillary microcavity wash-in
It washs clean；
S3：To being passed through dense H in the capillary microcavity after S2 step is washed2SO4Hydrogen peroxide solution, in the technical program
In, concentrated sulfuric acid volume accounting is 95%, and liquid is sealed in capillary microcavity and stands 1 hour, guarantees capillary microcavity inner wall
Hydroxyl on surface sufficiently activates, after by washes clean in capillary microcavity, then keep capillary microcavity inner hollow,
40 DEG C of freeze-day with constant temperature；
S4：3. aminopropyl triethoxysilane solution are passed through in capillary microcavity, stand 30 minutes, with deionized water and
Ethanol wash removes the silane compound of non-covalent bonding in capillary microcavity；
S5：The PBS solution of the antibody containing CD4+ is passed through in capillary microcavity, stands 1 hour, so that the cultivation of CD4+ antibody is arrived
The good capillary microcavity inner wall surface of silanization, after, it is successively passed through alcohol and deionized water into capillary microcavity, cleans
Inside capillary microcavity；
S6：1XCD4+T cell PBS solution to be measured is passed through capillary microcavity, records the Whispering-gallery-mode of different time
Resonance spectrum, specifically, every Whispering-gallery-mode resonance spectrum of 3 minutes records, until CD4+T cell is completely combined with antibody；
In S6 step, the time that CD4+T cell and antibody are completely combined is 30~40 minutes.
S7：Deionized water is passed through to the capillary microcavity after S6 step process and PBS solution is cleaned, is passed through again later
The PBS solution of 10XCD4+T cell records the echo wall die resonance spectrum of different time, specifically, primary every three minutes records
Echo wall die resonance spectrum, until CD4+T cell is completely combined with antibody, echo wall die resonance spectrum no longer drifts about.
Photobiology sensing is realized by the control and interaction of light and microfluid in capillary microcavity, and microcavity has small
Diameter forms the totally-enclosed of lymphocyte solution, micro bearer path, realizes analyte channel in conjunction with microflow control technique
With the separation of detection channels, measuring stability is improved.
After implementing above-mentioned S5 step, i.e., Fig. 4 is：The PBS solution of the antibody containing CD4+ is passed through time that test in microcavity obtains
Sound wall mode resonance Frequency bias changes over time situation, and within the certain effect time, antibody is attached to the interior inner surface of microcavity, by
It is thickeied in wall thickness, also there is a phenomenon where drift about for Whispering-gallery-mode resonance spectrum.
Fig. 5 is to implement above-mentioned S6 step, and different time is surveyed after the PBS solution containing 1XCD4+T cell is passed through into microcavity
The Whispering-gallery-mode resonance spectrum obtained is tried, within the certain effect time, CD4+T cell antigen and CD4+ antibody cell are combined, and are returned
Sound wall mode resonance spectrum is drifted about, and is tended towards stability after 30 minutes.Fig. 6 is the wave length shift of Whispering-gallery-mode resonance spectrum
Change with time situation, and wave length shift about 10pm, i.e. sensitivity reach 10pm/ (1.66X10-15mol/L)。
Fig. 7 is to implement above-mentioned S7 step, and different time is surveyed after the PBS solution containing 10XCD4+T cell is passed through into microcavity
The Whispering-gallery-mode resonance spectrum obtained is tried, after certain reaction time, CD4+T cell antigen and CD4+ antibody cell are combined, and are returned
Sound wall mode resonance spectrum is drifted about, and is tended towards stability after 20 minutes.Fig. 8 is the wave length shift of Whispering-gallery-mode resonance spectrum
Change with time situation, and wave length shift about 14pm, i.e. sensitivity reach 14pm/ (1.66X10-14Mol/L), and it is real
The detection of unmarked CD4+T lymphocyte is showed.
On the one hand the sensor utilizes echo in the evanscent field coupling process of micro-nano fiber and capillary microcavity excitation microcavity
Wall mode resonance, Whispering-gallery-mode resonance effects enhances microcavity inner wall area, and nearby light field and the lymphocyte inside microcavity are molten
The intensity of specific biological optics interaction occurs for liquid, can be quick, highly sensitive by the variation of Whispering-gallery-mode resonance spectrum
Tested lymphocyte characteristic is demodulated, interaction process time is short and is not necessarily to any fluorescent marker process.On the other hand, capillary
Microcavity has minute diameter, and by combining microflow control technique in test process, the totally-enclosed for foring lymphocyte solution is micro
Bearer path realizes analyte channel and detection channels and separates, realizes to the micro, high stability of lymphocyte solution
Photobiology test.The micro-nano fiber and capillary microcavity that the sensor uses have all -fiber, compact-sized, preparation is simple etc.
Feature.Therefore, the utility model is detected with high sensitivity, quickly, unmarked, compact-sized, integrated level is high, at low cost, work
The advantages that skill is simple.
Still there are many embodiment, all technologies formed using equivalents or equivalent transformation for the utility model
Scheme is all fallen within the protection scope of the utility model.
Claims (8)
1. a kind of lymphocyte Photobiology sensor based on optofluidic capillary microcavity, it is characterised in that：Swash including frequency sweep
Light device, capillary microcavity-micro-nano fiber coupling unit, photodetector and feedback for carrying out sensing testing to cell solution
Control system, the frequency swept laser, capillary microcavity-micro-nano fiber coupling unit, photodetector and feedback control system
It is of coupled connections between each other by fiber fuse, the feedback control system is electrical with photodetector and frequency swept laser respectively
Connection, the capillary microcavity-micro-nano fiber coupling unit are mutually perpendicular to be coupled to form by micro-nano fiber and capillary microcavity.
2. a kind of lymphocyte Photobiology sensor based on optofluidic capillary microcavity according to claim 1,
It is characterized in that：The wavelength of the frequency swept laser is tunable.
3. a kind of lymphocyte Photobiology sensor based on optofluidic capillary microcavity according to claim 1,
It is characterized in that：The micro-nano fiber is by single mode optical fiber by drawing cone machine fused biconical taper to be made.
4. a kind of lymphocyte Photobiology sensor based on optofluidic capillary microcavity according to claim 1,
It is characterized in that：The capillary microcavity carries out fused biconical taper by hollow quartz capillary optical fiber and is prepared.
5. a kind of lymphocyte Photobiology sensor based on optofluidic capillary microcavity according to claim 1,
It is characterized in that：The micro-nano fiber is close proximity on capillary microcavity, and the central axes of the micro-nano fiber and capillary microcavity are kept
Antarafacial is vertical.
6. a kind of lymphocyte Photobiology sensor based on optofluidic capillary microcavity according to claim 1,
It is characterized in that：The capillary microcavity is high symmetrical hollow cylinder shape structure.
7. a kind of lymphocyte Photobiology sensor based on optofluidic capillary microcavity according to claim 1,
It is characterized in that：The resonance wavelength for only meeting Whispering-gallery-mode resonance condition can generate resonance in capillary microcavity, full
The resonance wavelength of sufficient Whispering-gallery-mode resonance condition is determined by following formula：
The π of λ=2 rneff/m
Wherein r is resonant cavity radius, neffIt is the effective refractive index that resonant optical mode passes through, m is integer.
8. a kind of lymphocyte Photobiology sensor based on optofluidic capillary microcavity according to claim 1,
It is characterized in that：Resonant cycle period of the light wave in capillary microcavity determine by the quality factor q of capillary microcavity, useful effect
Length LeffIt is given by with quality factor q relationship：
Leff=Q the π of λ/2 neff。
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110308080A (en) * | 2019-07-23 | 2019-10-08 | 电子科技大学 | Utilize the method for functionalized capillary measurement carboxyl polystyrene microsphere concentration |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110308080A (en) * | 2019-07-23 | 2019-10-08 | 电子科技大学 | Utilize the method for functionalized capillary measurement carboxyl polystyrene microsphere concentration |
| CN110308080B (en) * | 2019-07-23 | 2021-04-06 | 电子科技大学 | Method for measuring concentration of carboxyl polystyrene microspheres by using functionalized capillary tube |
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