CN109507162A - A kind of laser detection system and method based on resonant cavity and FRET effect - Google Patents
A kind of laser detection system and method based on resonant cavity and FRET effect Download PDFInfo
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- CN109507162A CN109507162A CN201811515899.5A CN201811515899A CN109507162A CN 109507162 A CN109507162 A CN 109507162A CN 201811515899 A CN201811515899 A CN 201811515899A CN 109507162 A CN109507162 A CN 109507162A
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- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
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- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
A kind of laser detection system and method based on resonant cavity and FRET effect, including laser light source module, resonant tank module, optical collimating module and signal receiving module, laser light source module is used to for laser source to be coupled to resonant tank module and collects the signal light that resonant tank module generates in spectrometer for providing laser source, optical collimating module；Resonant tank module is for generating optical resonance and FRET effect, the microcavity including that can generate optical resonance, and the fluorescent receptor substance of FRET effect can be generated around microcavity；Signal receiving module is for receiving, analyzing, storing signal light.Laser light source module generates laser, laser source is coupled to resonant tank module by optical collimating module, excitation microcavity generates resonance, and then the fluorescent receptor substance around microcavity is excited to generate FRET effect, generates signal light；If the signal light and standard signal light that obtain have specific difference, show that testing molecule exists, realizes the detection of the outer micro substance of resonant cavity.
Description
Technical field
Present invention is primarily based on the optical resonators of Whispering-gallery-mode and fluorescence resonance energy transfer (fluorescent
Resonance energy transfer, FRET) effect.Pumping excitation is carried out to fluorescence beads with specific laser source, bead produces
Raw cluster laser simultaneously excites the fluorescent media outside bead by FRET effect, and system exports the laser of another cluster adjacent band, leads to
The frequency displacement of two beam laser wave spectrums of measurement is crossed, realizes and the hypersensitivity of biomolecule is sensed.
Background of invention
The Echo Wall (Whispering Gallery mode, WGM) phenomenon is all deposited in Beijing the Temple of Heaven and London St Paul's
, be primarily due to sound wave be bent smooth metope constantly reflect and be lost it is extremely low, so speak in wall side certain point, metope
Far another point remains to hear.The Echo Wall principle of optical mode is similar, the glass made of quartz material molten sintering
Glass pearl is exactly the Echo Wall microcavity of most original, and since bead inner boundary degree of roughness is small, inside and outside refractive index difference is larger, beats
Light beam into bead is easy to be not easy to be scattered absorption in intracavitary generation total reflection.Because of the wave band phase of bead size and light
Closely, photon can form the simple microlaser of Low threshold in intracavitary resonance.The ingredient of resonant cavity is from all kinds of silica-base materials
Or crystal expands to semiconductor and polymer, and can possess diversified cavity mold formula, such as spherical, micro- dish-type, twin columns capel
Deng.The advantages such as size is small, various structures, preparation are simple make the microcavity of Whispering-gallery-mode attract more and more researchers.
Compared with traditional Fabry-Perot-type cavity, Echo Wall chamber it is smaller, it is easier to it is integrated, since itself has pole
High quality factor, and size can accomplish from hundreds of nanometers to 500 μm to differ, the microcavity of different refractivity and absorptivity has
Different characteristics, before there is extraordinary application in the fields such as optical sensing, filter delayer, nonlinear optics and bio-imaging
Scape.For example, will be observed output variation by small sample perturbations, Echo Wall bead can when the size of optical cavity is other in the micron-scale
Using as extremely sensitive optics fine motion sensing element.Coupled system based on waveguide and microcavity can couple and chamber resonance wavelength
Consistent signal, so Echo Wall bead can play the role of filter.Whispering-gallery-mode microcavity is even more in life in recent years
Object photonics are shown up prominently, and F.Vollmer in 2002 etc. realizes the detection to protein using microsphere resonator, and 2008
Year, his team realized the detection to single virus using Echo Wall bead again.The Seok Hyun Yun of Harvard University in 2011
It takes the lead in realizing unicellular organism laser, followed by researcher by Echo Wall theory and cell combination, it is found that cell can be with
Phagocytosis with oneself itself the comparable Echo Wall bead of size, and can normal existence be metabolized one week or so up to procedural apoptosis.It is logical
Cell-Echo Wall feedback light is crossed, available miscellaneous optical data is simultaneously analyzed.It can be seen that Echo Wall chamber
It has a high potential in optical bio photonics, the design of integrated microfluidic platform can be more focused on to this research work in the future.
FRET effect refers to that when distance is in 1~10nm excitation energy is conducted from donor molecule (D) to acceptor molecule (A)
Phenomenon.For some specific Donor Acceptor pair, energy lost in donor is exactly the obtained energy of receptor.FRET effect
Rate ktThe distance between dipole r meets formulaWherein coefficient τ0By the spectral characteristic of dipole itself
It determines, R0Refer to the distance between dipole when FRET efficiency reaches 50%.Now, FRET has been few in number can be realized
One of several technological means of nanoscale distance detection, this also depends on novel fluorescence probe dye and the day of continuous development certainly
The algorithm of benefit optimization.
Summary of the invention
The present invention utilizes miniature optical resonant cavity, by FRET principle, realizes the detection of the outer micro substance of resonant cavity.In quilt
In the presence of detecting substance, the laser generated with the micro resonant cavity of light source pumping fluorescence doping can be excited micro- by FRET effect
Another fluorescent material (fluorescent receptor) outside ball makes it generate laser output.By adjusting the concentration of fluorescent dopants matter,
Two beam laser may be implemented while radiating.The wave spectrum of two-beam can be subjected to displacement with the variation of determinand, realize accordingly to
Survey the sensing detection of object.
The Echo Wall bead adulterated using laser 473nm light source activation dyestuff Dragon Green (DG), bead are excited
The fluorescence peak of 500~600nm wavelength out, after further increasing light source power, reach can generate 520 after resonant cavity threshold value~
The cluster laser peak of 540nm wave band.Energy part in bead can be transferred to outside ball by we using FRET effect, continue by
The outer liquid dissolved with fluorescent dye of bead excites, and generates new fluorescence peak in adjacent not far long-wave band, or reaching threshold value
The cluster laser peak between 540~570nm is generated afterwards, and experiment has verified that invention described above.Theory can be predicted at other
Wave band can also generate similar " spectrum migration " effect.
It is a primary object of the present invention to construct an optical system and method based on microcavity, passed for accurate biology
Sense.The system includes laser light source module (1), resonant tank module (2), optical collimating module (3) and signal receiving module (4),
In, laser light source module (1) is for providing laser source, including laser source (11), continuous attenuator (12)；Optical collimating module (3)
Spectrometer is collected for laser source to be coupled to resonant tank module (2), and by the signal light that resonant tank module (2) generate
In, including the microcobjective (31) (32) that two pieces of focal lengths are overlapped, dichroscope (33), convex lens (34) and camera (35)；It is humorous
Vibration cavity mold block (2) is for generating optical resonance and FRET effect, the microcavity including that can generate optical resonance, can generate around microcavity
The fluorescent receptor substance and three-dimensional trim holder of FRET effect；Signal receiving module (4) is for receiving, analyzing, storing signal
Light, including filter plate (41), convex lens (42), spectrometer (43)；The output laser of laser source (11) is successively by continuous decaying
Piece (12), dichroscope (33), microcobjective (31), resonant tank module (2) produce signal light, and signal light is divided into two parts,
A part of signal light successively passes through microcobjective (32), filter plate (41), convex lens (42), finally enters spectrometer (43) progress
It receives, analysis, storage, another part signal light successively passes through microcobjective (31), dichroscope (33), convex lens (34) and takes the photograph
As head (35), according to the image observed in real time, use three-dimensional trim holder for observing the coupling effect of laser and resonant cavity in real time
It adjusts microcavity position and realizes Best Coupling.
The microcobjective (31) (32) that two pieces of focal lengths are overlapped can also be aobvious by two pieces of convex lens replacements or one piece
Speck mirror and one piece of convex lens replacement.
The camera (35) can be CCD or cmos image sensor or blank.
The microcavity material can be quartz, doping quartz, polymer or the material for being capable of forming optical resonance.
The form of the microcavity can be microballoon, micro- disk, micro-loop, micro-pipe, micro- polygon, and geometric radius should be greater than swashing
The single wavelength dimension of light source (1) is encouraged, and less than 1 millimeter.
Fluorescent material can be added in the microcavity, or is not added.
The outer fluorescent receptor substance of the chamber can be gaseous state, liquid or solid-state, and its refractive index is less than in resonant cavity
Refractive index；The outer fluorescent receptor substance of the chamber may include multiple fluorescent substance, for multiple FRET effect to occur.
The resonant tank module (2) can transform micro-fluidic structure as.
The resonant frequency of exciting light source module (1) is identical as the native resonant frequency of resonant tank module (2).
A kind of laser detecting method based on resonant cavity and FRET effect, laser light source module (1) generates laser, by optics
Laser source is coupled to resonant tank module (2) by collimating module (3), and excitation microcavity generates resonance, and then excites glimmering around microcavity
Light receptor substance generates FRET effect, generates signal light；
If specific difference is not present in the signal light and standard signal light that obtain, show that testing molecule is not present；
If the signal light and standard signal light that obtain have specific difference, show that testing molecule exists；
Wherein, the standard signal light is to contain only the preset fluorescent receptor object of system in the medium around microcavity
When matter, the signal light that FRET effect generates occurs.
Beneficial effect
Compared with prior art, the present invention has the advantage that
1, the normal physiological context of destruction cell is generally required for detecting the conventional method of biological enzyme, such as cell
Certain enzymatic activity observation, it usually needs first clasmatosis processing reaction substrate is added thereto again, pass through observation substrate
Concentration variation carrys out reaction enzymes catalytic effect.According to the present invention, Echo Wall optical mode is combined with FRET effect, Ke Yishi
To the movable monitoring of protein molecular in the case where present cell normal growth metabolism, analyzing spectral signature can be obtained closer to thin
The physiological characteristic of the enzyme of born of the same parents' nature.
2, for nuclear structure of the present invention in micron-scale, material used is the stable plastic polymer of performance or quartzy glass
Glass, it can be used repeatedly.In conjunction with the light source and micro-nano lens design technique of present maturation, more easily system can be miniaturized
Even it is made into system on chip.
3, single strand dna and immune protein molecule are generally being no more than nano-scale, and having exceeded optical microscopy can
The range of observation.It is previous it is most of about DNA hybridization and the method for immunoassay be all that isotope labelling or molecular engram are shown again
Picture, our microcavity sensor-based system can detect the combination or conformation change of molecule within the scope of several microns, for research
DNA molecular hybridization and immunoassay provide new method.
4, Whispering-gallery-mode is combined with FRET with distinctive advantage.Although being detected using Whispering-gallery-mode
There are many reports, but the detection system being applicable in is often fairly simple.And combine Whispering-gallery-mode and FRET, using suitable
When fluorochrome label be detected molecule, the specific detection of target molecule in complex system may be implemented.
As a whole, laser sensing detection system based on Whispering-gallery-mode and FRET principle biomolecule dynamics,
Immunoassay, detection of nucleic acids and protein-protein interaction suffer from huge potentiality and application.About cell life generation
The research for journey of apologizing for having done sth. wrong has extremely important status in biology, and the present invention is that observation is new this process provides one in real time
Approach.
Detailed description of the invention:
Fig. 1 is laser sensing detecting system schematic diagram of the embodiment 1 based on Whispering-gallery-mode and FRET principle
Fig. 2 is laser sensing detecting system schematic diagram of the embodiment 2 based on Whispering-gallery-mode and FRET principle
Fig. 3 is laser sensing detecting system schematic diagram of the embodiment 3 based on Whispering-gallery-mode and FRET principle
Fig. 4 is laser sensing detecting system schematic diagram of the embodiment 4 based on Whispering-gallery-mode and FRET principle
Fig. 5 is laser sensing detecting system schematic diagram of the embodiment 5 based on Whispering-gallery-mode and FRET principle
Fig. 6 is the appearance signal of resonant cavity sample in the laser sensing detection system based on Whispering-gallery-mode and FRET principle
Figure
Fig. 7 is the FRET effect detailed schematic of the laser sensing detection system based on Whispering-gallery-mode and FRET principle
Fig. 8 is the microcosmic schematic diagram of FRET effect of the laser sensing detection system based on Whispering-gallery-mode and FRET principle
Fig. 9 a-d is the signal spectrum schematic diagram of the laser sensing detection system based on Whispering-gallery-mode and FRET principle
Wherein, laser source (11), continuous attenuator (12), silver mirror (13), resonant cavity sample (21), dichroscope (33),
Convex lens (34), filter plate (41), convex lens (42), spectrometer (43)
Specific embodiment
Below with reference to embodiment, the present invention will be further described, but the present invention is not limited to following embodiments.
Case study on implementation 1
As shown in Figure 1, resonant cavity sample (21) uses 15 μm of average diameter or so of doping dragon green dyestuff
Polymer globules (Bangs Laboratories, FS07F), take bead original solution to be added to prepared rhodamine in right amount
(RhB) among solution, reagent is dripped on the cover slip after being sufficiently mixed uniformly, is covered on above with another coverslip, and
Surrounding is smeared into nail polish encapsulation process (Fig. 6).Above-mentioned sample is fixed on glass slide after place to be sealed is dry, is thus made
It has been made the sample to be tested that can be placed on precision three-dimensional adjustment frame.
Continuous sheet of attenuator (12) are housed after laser source (11), after (13) 90 ° of a piece of silver mirror reflections, light beam is again
90 ° are reflected by the high anti-dichroscope (33) of the low pass of a piece of 500nm, light beam is made to be focused standard by 40 × microcobjective (31)
Directly, it is coupled on the resonant cavity sample (21) being mounted on precision three-dimensional adjustment frame.The another side of dichroscope (33) is equipped with and takes the photograph
As head (35).The rear end of resonant cavity sample (21) is equipped with 60 × micro objective (32), 500nm long pass filter piece (41) and focal length
F=18.40mm convex lens (42), into the probe of spectrometer (43).Experimental provision takes the positive mode detected, in dichroic
The convex lens (34) of a piece of focal length f=100mm is assembled in the another side of mirror (33) and camera (35) carries out in real time small ball position
It observes (Fig. 7).
Sample is placed on vertically in the optical path between two microcobjectives when detection.Continuous attenuator (12) is adjusted by light
The light intensity regulating in source observes 473nm by the CCD camera of the other end (35) to lower intensity, constantly adjustment sample position
Whether the laser source (11) of wavelength is coupled on bead, until bead output light shown on spectrometer it is most strong until (Fig. 8).
Then ever-increasing laser coupled injection is carried out to the same bead, since bead has very strong Echo Wall effect, when
473nm laser coupled is into the fluorescence that fluorescent material dragon green in ball after bead, can be excited to issue 500~600nm wave band.
Cluster laser peak (Fig. 9 a) will be issued after 473nm laser source strength reaches bead threshold value, in the fluorescent material of debita spissitudo
FRET effect will be generated among solution RhB, this cluster laser peak is transferred to another wave band f1(energy is shifted to longer wave band
Amount) (Fig. 9 b).On the basis of the above, testing molecule is added around bead, just when testing molecule is less than 10nm apart from bead
Meeting generates disturbance to the FRET effect of script, forms new FRET spectrum f2(Fig. 9 c).Since different molecular makes sensor-based system
At disturbance it is different, generated spectral signature is not also identical, and the spectrum change for comparing disturbance front and back being capable of specific detection
To the presence or absence and motion conditions of the molecule.
Case study on implementation 2
As shown in Fig. 2, the silver mirror (13) in embodiment 1 is removed, the light-emitting angle of laser source (11) is adjusted to vertical
In on the direction of optical alignment system.Other execution methods are in the same manner as in Example 1, can be realized same detection effect.
Case study on implementation 3
As shown in figure 3, the silver mirror (13) in embodiment 1 is removed, the light-emitting angle of laser source (11) is adjusted to vertical
In on the direction of optical alignment system.40 × microcobjective (31) and 60 × micro objective (32) are replaced into convex lens,
It is in the same manner as in Example 1 that he executes method, can be realized same detection effect.
Case study on implementation 4
As shown in figure 3, the silver mirror (13) in embodiment 1 is removed, the light-emitting angle of laser source (11) is adjusted to vertical
In on the direction of optical alignment system.40 × microcobjective (31) and 60 × micro objective (32) are replaced into convex lens, it will
Camera (35) is replaced into blank, other execution methods are in the same manner as in Example 1, can be realized same detection effect.
Case study on implementation 5
As shown in figure 5, microfluidic channel (21) can accurately control the flowing of solution.This sensor-based system uses average diameter
The polymer globules (Bangs Laboratories, FS07F) of 15 μm or so of doping dragon green dyestuff, take small first
Ball original solution is added among prepared rhodamine (RhB) solution in right amount and is uniformly mixed, then mixed solution is injected into miniflow
It controls in channel (21).
Continuous sheet of attenuator (12) are housed after laser source (11), light beam by a piece of 500nm the high anti-dichroic of low pass
Mirror (33) reflects 90 °, and light beam is made to be focused collimation by 40 × microcobjective (31), is coupled to and is mounted in precision three-dimensional adjustment
On microfluidic channel (21) on frame.The another side of dichroscope (33) is equipped with camera (35).After resonant cavity sample (21)
End is equipped with 60 × micro objective (32), 500nm long pass filter piece (41) and focal length f=18.40mm convex lens (42), into light
The probe (43) of spectrometer.Experimental provision takes the positive mode detected, assembles a piece of focal length f in the another side of dichroscope (33)
The convex lens (34) of=100mm and camera (35) for imaging observe bead coupling effect in real time.
The FRET effect spectrum f1 of record system generation first.On the basis of the above, it is added in microfluidic channel to be measured
Molecule, when testing molecule is less than 10nm apart from bead will FRET effect to script generate disturbance, form new FRET light
Compose f2.Comparing the difference between f1 and f2 can be achieved with specific molecular detection.
Case study on implementation 6
As shown in Figure 1, the resonant tank module (2) in embodiment 1 is slightly modified, other equipment are in the same manner as in Example 1,
It can be realized the monitoring to cellular physiological events.Specific transformation are as follows: for a period of time by cell and bead co-incubation, bead will
It is swallowed by cell, and cell normal existence and metabolism.The cell of phagocytosis bead is taken to be added to prepared rhodamine in right amount
(RhB) among solution, reagent is dripped on the cover slip after being sufficiently mixed uniformly, is covered on above with another coverslip, and
Surrounding is smeared into nail polish encapsulation process.Above-mentioned sample is fixed on glass slide after place to be sealed is dry, is thus fabricated to
The resonant cavity sample (21) that can be placed on precision three-dimensional adjustment frame.
Resonant cavity sample (21) is placed in the optical path between two microcobjectives vertically when detection.Adjust continuous decaying
The light intensity regulating of light source to lower intensity, constantly adjustment sample position are passed through the CCD camera of the other end (35) by piece (12)
Whether the laser source (11) of observation 473nm wavelength couples on bead in the cell, until the light of bead output is on spectrometer
Until display is most strong.Then ever-increasing laser coupled injection is carried out to the same bead, since bead has very strong time
Sound wall effect can excite fluorescent material dragon green in ball to issue 500~600nm after 473nm laser coupled is into bead
The fluorescence of wave band.Cluster laser peak will be issued after 473nm laser source strength reaches bead threshold value.It also can be because in cytoplasm
For absorption, there are fluorescent receptor substance RhB, so intracellular bead will generate FRET effect, by this cluster laser
Peak is transferred to another wave band f1 (Fig. 9 b).On the basis of the above, when cell metabolism, the molecule in cytoplasm is in bead week
Enclose continuous movement, when it is less than 10nm at a distance from bead will FRET effect to script generate disturbance, formed new
FRET spectrum f2 (Fig. 9 c).Since different molecular disturbs difference caused by sensor-based system, generated spectral signature also not phase
Together, the spectrum change for comparing disturbance front and back is capable of the physiological activity of specific detection cell.
Case study on implementation 7
As shown in figure 5, the resonant tank module (2) in embodiment 5 is slightly modified, other equipment are in the same manner as in Example 5,
It can be realized the movable monitoring of hybridization single-stranded to DNA.Specific transformation are as follows: by Echo Wall bead surface progress sulfhydrylation and altogether
Single stranded DNA is fixed to bead surface by the method for valence bonding reaction.It can be with by imaging tools such as super-resolution Laser Scanning Confocal Microscopes
Determine the DNA quantity of modification, controls the conditions such as reaction density and time, the modification of one or several chain is realized on bead.?
After fixed dna is single-stranded on the outside of Echo Wall bead, it is put into microchannel.Laser light source and coupling position are adjusted to progress
It excites, a certain concentration substance such as quantum dot that FRET effect can occur is contained in fluid solution.Measurement note after the system stabilizes
Record spectral information.When being slowly passed through containing the single-stranded solution of DNA to be measured, this anchor chain can hybridize with the anchoring chain on bead
Reaction.Due to double-stranded DNA in structure it is more more compact than single stranded DNA securely, hybridization combine after DNA can occur outside ball shape with
Variation on direction, and then interference can be generated to the microenvironment outside the ball of bead.Since liquid environment generates outside bead and ball
The condition of FRET is very sensitive, and interference brought by this external environment variation can be intuitively shown in the variation of spectrum.It utilizes
This detection means, we can be realized the nucleic acid molecules detection of distinguished sequence.
Claims (10)
1. a kind of laser detection system based on resonant cavity and FRET effect, it is characterised in that: the system includes laser light source module
(1), resonant tank module (2), optical collimating module (3) and signal receiving module (4), wherein laser light source module (1) is for providing
Laser source, including laser source (11), continuous attenuator (12)；Optical collimating module (3) is used to laser source being coupled to resonant cavity
Module (2), and the signal light that resonant tank module (2) generate is collected in spectrometer, it is overlapped including two pieces of focal lengths micro-
Object lens (31) (32), dichroscope (33), convex lens (34) and camera (35)；Resonant tank module (2) is for generating optical resonance
With FRET effect, the microcavity including that can generate optical resonance can generate the fluorescent receptor substance of FRET effect around microcavity, with
And three-dimensional trim holder；Signal receiving module (4) is aspherical for receiving, analyzing, storing signal light, including filter plate (41)
Mirror (42), spectrometer (43)；The output laser of laser source (11) successively passes through continuous attenuator (12), dichroscope (33), shows
Speck mirror (31), resonant tank module (2) produce signal light, and signal light is divided into two parts, and a part of signal light is successively by aobvious
Speck mirror (32), filter plate (41), non-spherical lens (42) finally enter spectrometer (43) and are received, analyzed, stored, separately
A part of signal light successively passes through microcobjective (31), dichroscope (33), convex lens (34) and camera (35), for real-time
The coupling effect for observing laser and resonant cavity is realized according to the image observed in real time with three-dimensional trim holder adjustment microcavity position
Best Coupling.
2. a kind of laser detection system based on resonant cavity and FRET effect, feature according to claims 1 exist
In: the microcobjective (31) (32) that two pieces of focal lengths are overlapped can also be by two pieces of convex lens replacements or one piece of micro- object
Mirror and one piece of convex lens replacement.
3. a kind of laser detection system based on resonant cavity and FRET effect, feature according to claims 1 exist
In: the camera (35) can be CCD or cmos image sensor or blank.
4. a kind of laser sensing detection system based on microsphere resonator and FRET effect according to claims 1,
It is characterized by: the microcavity material can be quartz, doping quartz, polymer or the material for being capable of forming optical resonance.
5. a kind of laser detection system based on resonant cavity and FRET effect, feature according to claims 1 exist
In: the form of the microcavity can be microballoon, micro- disk, micro-loop, micro-pipe, micro- polygon, and geometric radius should be greater than excitation light
The single wavelength dimension in source (1), and less than 1 millimeter.
6. a kind of laser detection system based on resonant cavity and FRET effect, feature according to claims 1 exist
In: fluorescent material can be added in the microcavity, or is not added.
7. a kind of laser detection system based on resonant cavity and FRET effect, feature according to claims 1 exist
In: the outer fluorescent receptor substance of the chamber can be gaseous state, liquid or solid-state, and its refractive index is less than the refraction in resonant cavity
Rate；The outer fluorescent receptor substance of the chamber may include multiple fluorescent substance, for multiple FRET effect to occur.
8. according to the laser detection system based on resonant cavity and FRET effect a kind of in claims 1, it is characterised in that: described
Resonant tank module (2) micro-fluidic structure can be transform as.
9. a kind of laser detection system based on resonant cavity and FRET effect, feature according to claims 1 exist
In: the resonant frequency of exciting light source module (1) is identical as the native resonant frequency of resonant tank module (2).
10. a kind of laser detecting method based on resonant cavity and FRET effect, it is characterised in that: laser light source module (1), which generates, to swash
Laser source is coupled to resonant tank module (2) by light by optical collimating module (3), and excitation microcavity generates resonance, and then excites micro-
Fluorescent receptor substance around chamber generates FRET effect, generates signal light；
If specific difference is not present in the signal light and standard signal light that obtain, show that testing molecule is not present；
If the signal light and standard signal light that obtain have specific difference, show that testing molecule exists；
Wherein, the standard signal light is to contain only the preset fluorescent receptor substance of system in the medium around microcavity
When, the signal light that FRET effect generates occurs.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110208214A (en) * | 2019-06-24 | 2019-09-06 | 福建师范大学 | A kind of hydroscope excitation microcavity laser sensor-based system and method |
| CN111999270A (en) * | 2020-06-10 | 2020-11-27 | 北京工业大学 | Exosome detection method based on whispering gallery mode optical microcavity |
| CN112625900A (en) * | 2020-12-17 | 2021-04-09 | 西安电子科技大学 | Electromagnetic radiation cell experimental device with inclined waveguide resonant cavity |
| CN112630198A (en) * | 2020-09-15 | 2021-04-09 | 北京工业大学 | Sensing detection method based on whispering gallery mode optical microcavity |
| CN112763463A (en) * | 2020-11-05 | 2021-05-07 | 北京工业大学 | Method for improving sensitivity of whispering gallery mode microcavity sensing |
| CN115166960A (en) * | 2022-07-19 | 2022-10-11 | 华南师范大学 | LED excitation light source for quantitative FRET microscopic imaging and dimming method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102980853A (en) * | 2012-11-23 | 2013-03-20 | 天津大学 | Resonant light microfluid sensing device and method based on active micropipe |
| CN104868352A (en) * | 2015-06-03 | 2015-08-26 | 吉林大学 | Three-dimensional asymmetric miniature resonant cavity polymer single-mode laser |
| WO2017210675A1 (en) * | 2016-06-03 | 2017-12-07 | The General Hospital Corporation | System and method for micro laser particles |
| CN108574199A (en) * | 2018-04-18 | 2018-09-25 | 云南大学 | A kind of fluorescence resonance energy transfer optofluidic laser and its preparation method and application |
-
2018
- 2018-12-11 CN CN201811515899.5A patent/CN109507162B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102980853A (en) * | 2012-11-23 | 2013-03-20 | 天津大学 | Resonant light microfluid sensing device and method based on active micropipe |
| CN104868352A (en) * | 2015-06-03 | 2015-08-26 | 吉林大学 | Three-dimensional asymmetric miniature resonant cavity polymer single-mode laser |
| WO2017210675A1 (en) * | 2016-06-03 | 2017-12-07 | The General Hospital Corporation | System and method for micro laser particles |
| CN109496378A (en) * | 2016-06-03 | 2019-03-19 | 通用医疗公司 | System and method for micro laser particle |
| CN108574199A (en) * | 2018-04-18 | 2018-09-25 | 云南大学 | A kind of fluorescence resonance energy transfer optofluidic laser and its preparation method and application |
Non-Patent Citations (3)
| Title |
|---|
| MATJAŽ HUMAR ET AL.: "Intracellular microlasers", 《NATURE PHOTONICS》 * |
| SIYKA I. SHOPOVA等: "Opto-fluidic ring resonator lasers based on highly efficient resonant energy transfer", 《OPTICS EXPRESS》 * |
| VENKATA RAMANAIAH DANTHAM等: "Influence of whispering gallery modes (WGMs) on energy transfer", 《CHEMICAL PHYSICS》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110208214A (en) * | 2019-06-24 | 2019-09-06 | 福建师范大学 | A kind of hydroscope excitation microcavity laser sensor-based system and method |
| CN111999270A (en) * | 2020-06-10 | 2020-11-27 | 北京工业大学 | Exosome detection method based on whispering gallery mode optical microcavity |
| CN112630198A (en) * | 2020-09-15 | 2021-04-09 | 北京工业大学 | Sensing detection method based on whispering gallery mode optical microcavity |
| CN112763463A (en) * | 2020-11-05 | 2021-05-07 | 北京工业大学 | Method for improving sensitivity of whispering gallery mode microcavity sensing |
| CN112625900A (en) * | 2020-12-17 | 2021-04-09 | 西安电子科技大学 | Electromagnetic radiation cell experimental device with inclined waveguide resonant cavity |
| CN115166960A (en) * | 2022-07-19 | 2022-10-11 | 华南师范大学 | LED excitation light source for quantitative FRET microscopic imaging and dimming method |
| CN115166960B (en) * | 2022-07-19 | 2023-08-04 | 华南师范大学 | LED excitation light source for quantitative FRET microscopic imaging and dimming method |
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