CN108801863A - The femtosecond optical optical tweezers system of colloidal particle dynamics and image-forming information in solution can be obtained - Google Patents
The femtosecond optical optical tweezers system of colloidal particle dynamics and image-forming information in solution can be obtained Download PDFInfo
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- 238000012576 optical tweezer Methods 0.000 title claims abstract description 46
- 230000003287 optical effect Effects 0.000 title claims abstract description 34
- 239000002245 particle Substances 0.000 title claims abstract description 24
- 230000005284 excitation Effects 0.000 claims abstract description 31
- 238000000799 fluorescence microscopy Methods 0.000 claims abstract description 19
- 238000001514 detection method Methods 0.000 claims abstract description 14
- 238000000879 optical micrograph Methods 0.000 claims abstract description 8
- 239000007850 fluorescent dye Substances 0.000 claims abstract description 6
- 239000000975 dye Substances 0.000 claims abstract description 5
- 210000004027 cell Anatomy 0.000 claims description 14
- 239000004793 Polystyrene Substances 0.000 claims description 11
- 229920002223 polystyrene Polymers 0.000 claims description 10
- 239000007787 solid Substances 0.000 claims description 10
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 6
- 239000010931 gold Substances 0.000 claims description 6
- 229910052737 gold Inorganic materials 0.000 claims description 6
- 238000003384 imaging method Methods 0.000 claims description 6
- 230000007935 neutral effect Effects 0.000 claims description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 4
- 238000013481 data capture Methods 0.000 claims description 4
- 210000005260 human cell Anatomy 0.000 claims description 4
- 238000005259 measurement Methods 0.000 claims description 4
- 239000008188 pellet Substances 0.000 claims description 4
- 229910052814 silicon oxide Inorganic materials 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 3
- 239000011324 bead Substances 0.000 description 8
- 239000000084 colloidal system Substances 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000003921 oil Substances 0.000 description 4
- 238000010586 diagram Methods 0.000 description 3
- 230000013011 mating Effects 0.000 description 3
- 238000012632 fluorescent imaging Methods 0.000 description 2
- 238000007654 immersion Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 241000276498 Pollachius virens Species 0.000 description 1
- RTAQQCXQSZGOHL-UHFFFAOYSA-N Titanium Chemical compound [Ti] RTAQQCXQSZGOHL-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000001246 colloidal dispersion Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 239000010419 fine particle Substances 0.000 description 1
- 238000002866 fluorescence resonance energy transfer Methods 0.000 description 1
- 229910052736 halogen Inorganic materials 0.000 description 1
- 150000002367 halogens Chemical class 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 230000009022 nonlinear effect Effects 0.000 description 1
- 239000013307 optical fiber Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 210000001747 pupil Anatomy 0.000 description 1
- 239000002096 quantum dot Substances 0.000 description 1
- 229910052594 sapphire Inorganic materials 0.000 description 1
- 239000010980 sapphire Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229910052719 titanium Inorganic materials 0.000 description 1
- 239000010936 titanium Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
-
- G01N15/01—
Abstract
The femtosecond optical optical tweezers system of colloidal particle dynamics and image-forming information in solution is obtained the present invention relates to a kind of, it is characterised in that the system includes：Stablize the femtosecond optical tweezer capture laser cell for capturing ligh trap for capturing laser as femtosecond and being provided for sample to be tested；Position detection unit for detecting captured sample to be tested position distribution when doing limited Brownian movement relative to ligh trap center；Excitation light source unit for carrying out fluorescence excitation as exciting light irradiation sample to be tested；For femtosecond capture laser to be collected into position detection unit, and the exciting light that excitation light source unit is emitted is introduced into the sample to be tested in capture ligh trap so that the luminescent dye molecule of sample to be tested, which is stimulated, generates the optical microphotograph unit of fluorescence signal；Fluorescence signal for collecting sample to be tested generation completes the real-time fluorescence imaging of single colloidal particle coated to fluorescent dye, obtains the single molecular fluorescence imaging unit of the dynamic information of single colloidal particle of the sample to be tested in different viscoelasticity characteristic systems.
Description
Technical field
The femtosecond optical optical tweezers system of colloidal particle dynamics and image-forming information in solution is obtained the present invention relates to a kind of, is related to
And colloid science and biophysics basic research field.
Background technology
Optical tweezer technology is a kind of important single molecule techniques that can be captured and manipulate fine particle and carry out mechanical meaurement,
Invented from the Ash gold of AT&T Labs of the U.S. in 1986, it is increasingly mature by development in more than 30 years, theory with
And application aspect has all done a large amount of research work, and dual access test, multiple light forceps, femtosecond optical tweezer and whirlpool are gradually developed by monochromatic light tweezer
All kinds of optical tweezers such as optically-active tweezer, it is more that this technology is widely used in biophysics, nanoprocessing, colloid science and physics etc.
A field, and optical tweezer is combined due to easy with some other technology, for example (,) single molecular fluorescence imaging, fluorescence resonance energy transfer
Deng becoming the big advantage relative to monotechnics or measurement means.Femtosecond optical tweezer is people by the capture light source of optical tweezer
It replaces continuous wave laser, femtosecond laser that there is the pulsewidth and high repetition frequency of femtosecond with femtosecond laser, it is made to meet continuously
While wave laser capture requires, it is also equipped with the property that continuous wave laser does not have, is achieved that under lower mean power
Capture.Cant woods et al. Lateral optical force for being put forward for the first time and calculating femtosecond optical tweezer in theory in 2004, the same year B.Agate
Et al. with femtosecond optical tweezer realize two-photon fluorescence excitation and in-situ control, find its capture needed for laser average intensity it is bright
It is aobvious to be less than continuous wave laser；Rainfall waits by force found that the non-linear capture due to strong-focusing femtosecond optical tweezer is formed in 2010 for the first time
The phenomenon that ligh trap cleaves；It is directly former in a micro volume that miniflows of the Goswami et al. based on new femtosecond optical tweezer becomes method
Position measures the temperature etc. at solid-liquid interface.Compared to traditional optical tweezer, the complex characteristics of femtosecond laser itself make femtosecond optical tweezer
Related Experimental Study it is all also very limited, therefore, by femtosecond optical tweezer with total internal reflection fluorescent imaging be combined, one kind can obtain molten
The foundation of the imaging measurement method for the HIGH SENSITIVITY AND HIGH PRECISION that the dynamic information of colloidal particle measures in liquid is to be highly desirable
's.
Invention content
In view of the above-mentioned problems, the object of the present invention is to provide one kind can obtaining colloidal particle dynamics and imaging letter in solution
The femtosecond optical optical tweezers system of breath, can to colloidal dispersion, cell carry out accurately actively manipulate while can also carry out high-resolution,
Highly sensitive unimolecule measures and imaging, and then obtains the dynamic information of colloidal particle or cell in the solution.
To achieve the above object, the present invention takes following technical scheme：One kind can obtain colloidal particle dynamics in solution
And the femtosecond optical optical tweezers system of image-forming information, the system include：For capturing laser as femtosecond stable catch is provided for sample to be tested
Obtain the femtosecond optical tweezer capture laser cell of ligh trap；For detecting captured sample to be tested limited Blang is being relative to ligh trap center
The position detection unit of position distribution when movement；Excitation light source for carrying out fluorescence excitation as exciting light irradiation sample to be tested
Unit；For femtosecond capture laser to be collected into the position detection unit, and the excitation that the excitation light source unit is emitted
Light is introduced into the sample to be tested in capture ligh trap so that the luminescent dye molecule of sample to be tested, which is stimulated, generates fluorescence signal
Optical microphotograph unit；Fluorescence signal for collecting sample to be tested generation completes single colloid grains coated to fluorescent dye
The real-time fluorescence imaging of son, obtains the dynamic information of single colloidal particle of the sample to be tested in different viscoelasticity characteristic systems
Single molecular fluorescence imaging unit.
Further, the optical microphotograph unit uses inverted fluorescence microscope structure, including microcobjective, the first~the
Three dichroscopes and condenser；Exciting light focuses on the sample to be tested in capture ligh trap through the microcobjective, through excitation
Light excitation sample to be tested generate fluorescence through the microcobjective collection after be emitted to the first dichroscope, through the described 1st to
The fluorescence signal of Look mirror outgoing is emitted to the single molecular fluorescence imaging measurement unit.
Further, femtosecond optical tweezer capture laser cell include femtosecond pulse laser, expand with collimating mirror and
Deflecting mirror expands described in the femtosecond pulse warp of femtosecond pulse laser outgoing and is expanded and collimated with collimating mirror,
Femtosecond pulse after expanding and collimating through the deflecting mirror into the second dichroscope is emitted to after horizontal deflection, through described second
The light emitting of dichroscope outgoing is to the microcobjective.
Further, the position detection unit includes convergent lens, positional detecting device, data collecting card and calculating
Machine；The interference pattern formed through the forward scattering light of microcobjective outgoing is emitted to the three or two after condenser collection
To Look mirror, the light through third dichroscope outgoing focuses the positional detecting device, the position through the convergent lens
Detection device connects the stiffness coefficient of ligh trap residing for the computer acquisition sample to be tested by the data collecting card and waits for
Viscosity of the sample in different viscoelastic systems.
Further, the positional detecting device uses Position-Sensitive Detector or digital camera.
Further, the excitation light source unit includes solid state laser, Glan-Taylor prism, quarter-wave plate, expansion
Beam and collimating mirror, lens and optical filter；The continuous wave that the solid state laser is sent out successively through the Glan-Taylor prism and
Quarter-wave plate be emitted to it is described expand and collimating mirror, through the circularly polarized light being emitted with collimating mirror that expands through successively through upper
The lens and optical filter focus on the back focal plane of the microcobjective.
Further, neutral density filter plate is arranged in the solid state laser exit.
Further, the single molecular fluorescence imaging unit uses EMCCD cameras and pci data capture card, the EMCCD
Camera connects the computer through the pci data capture card.
Further, the sample to be tested is that polystyrene fluorescence beads, gold nano grain, non-blooming polystyrene are small
Ball or silicon oxide pellets, Viable human cell.
The invention adopts the above technical scheme, which has the following advantages：1, the femtosecond of the invention by high repetition frequency
Pulse optical tweezer is combined with the imaging of unimolecule total internal reflection fluorescent, can obtain the dynamic information that system is measured in solution.
2, the present invention carries out exciting light using highly sensitive EMCCD cameras while can realize capture manipulation to sample to be tested
Excitation and the collection of fluorescence signal, and then realize to include two-photon fluorescence signal and the commonly detection of more fluorescence signals.3,
The sample to be tested of the present invention is colloid bead or gold nano grain etc. in different viscoelastic systems, therefore be may be implemented different viscous
Elastic system, various sizes of colloidal particle measure.Present invention can be extensively applied to nano particle, colloidal particle,
The dependent dynamics of a variety of systems such as gel, cell are studied, and the basic physics root of its respective special nature is explored.
Description of the drawings
Fig. 1 is the femtosecond optical optical tweezers system measuring principle schematic diagram of the present invention；
Fig. 2 is the femtosecond laser capture light source unit and excitation light source unit light path schematic diagram of the present invention；
Fig. 3 is the Displacements Distribution schematic diagram that the colloidal particle of the embodiment of the present invention moves in ligh trap；
Fig. 4 is the two-photon excitation image of the fluorescence PS beads of captured radius 100nm in the embodiment of the present invention.
Specific implementation mode
Come to carry out detailed description to the present invention below in conjunction with attached drawing.It should be appreciated, however, that attached drawing has been provided only more
Understand the present invention well, they should not be interpreted as limitation of the present invention.In the description of the present invention, it is to be understood that,
Term " first ", " second " etc. are only used for the purpose of description, are not understood to indicate or imply relative importance.
As shown in Figure 1, provided by the invention obtain colloidal particle dynamics and the femtosecond optical tweezer of image-forming information in solution
System, including femtosecond optical tweezer capture laser cell 1, position detection unit 2, excitation light source unit 3, optical microphotograph unit 4 and list
Molecular fluorescence imaging unit 5, wherein：
Femtosecond optical tweezer capture laser cell 1 is used to provide stable capture light as femtosecond capture laser for sample to be tested
Trap；
Position detection unit 2 is used to detect captured sample to be tested when doing limited Brownian movement relative to ligh trap center
Position distribution；
Excitation light source unit 3 is used to carry out fluorescence excitation as exciting light irradiation sample to be tested；
Optical microphotograph unit 4 is used to femtosecond capture laser being collected into position detection unit 2, and by excitation light source unit
The exciting light of 3 outgoing is introduced into the sample to be tested in capture ligh trap so that the luminescent dye molecule of sample to be tested is stimulated
Generate fluorescence signal；
Single molecular fluorescence imaging unit 5 is used to collect the fluorescence signal of sample to be tested generation, completes to be coated with fluorescent dye
Single colloidal particle real-time fluorescence imaging, obtain single colloidal particle of the sample to be tested in different viscoelasticity characteristic systems
Dynamic information；In addition, single molecular fluorescence imaging unit 5 can also be used for meeting the common light field of Kohler illumination condition at
Picture.
In a preferred embodiment, as shown in Figure 1, inverted fluorescence microscope knot may be used in optical microphotograph unit 4
Structure, including 40, three dichroscopes 41 of microcobjective, condenser 42, speculum 43 and diaphragm 44；Exciting light is through microcobjective 40
The sample to be tested in capture ligh trap is focused on, the fluorescence generated through excitation sample to be tested is collected through microcobjective 40
After be emitted to dichroscope 41c, the emitted smooth high-pass filtering piece of fluorescence signal through dichroscope 41c outgoing is emitted to speculum
43, the fluorescence reflected through speculum 43 is emitted to single molecular fluorescence imaging measurement unit 5 through diaphragm 44.
In a preferred embodiment, as shown in Fig. 2, femtosecond optical tweezer capture laser cell 1 includes providing femtosecond capture
The femtosecond pulse laser 10 of laser, is expanded and is filtered with collimating mirror 12, diaphragm 13, deflecting mirror 14 and two band logicals several speculums 11
Mating plate 15.TEM may be used in femtosecond pulse laser 1000The titanium sapphire optical fiber laser of pattern, femtosecond pulse laser 10
The femtosecond pulse of outgoing is reflected into expand through speculum 11a and is expanded and collimated with collimating mirror 12, after expanding and collimating
Femtosecond pulse be emitted to deflecting mirror 14 through diaphragm 13 and speculum 11b and 11c successively, through deflecting mirror 14 deflection outgoing
Light is emitted to dichroscope 41a through bandpass filter 15a, and the light through dichroscope 41a outgoing is emitted to through bandpass filter 15b
Microcobjective 40, pupil size after microcobjective 40 should be equal or slightly larger than by expanding rear beam diameter size, different numerical apertures it is micro-
Object lens 40 can slightly have difference, about 5~6.3mm, can ensure to obtain gradient force as big as possible in this way, to be advantageously implemented simultaneously
Keep stable capture, wherein the high-precision deflecting mirror of Piezoelectric Driving may be used in deflecting mirror 14, can be controlled by computer
It drives deflecting mirror 14 that microradian deflection occurs, realizes the active manipulation of femtosecond optical tweezer.
In a preferred embodiment, as shown in Figure 1, position detection unit 2 includes convergent lens 20, the filter of high pass low-resistance
Mating plate 21, Position-Sensitive Detector 22, NI data collecting cards 23 and computer 24；The forward scattering light being emitted through microcobjective 40
Through condenser 42, (condenser makes light beam preferably shine the component on sample, it is therefore an objective to as far as possible to the interference pattern of formation
Collection forward scattering light) collect after be emitted to dichroscope 41b, the concentrated lens successively of the light through dichroscope 41b outgoing
20 and 21 focal position sensing detector 22 of high pass low-resistance optical filter, Position-Sensitive Detector 22 passes through NI data collecting cards 23 and connects
Connect computer 24.It should be noted that Position-Sensitive Detector 22 could alternatively be digital camera, digital camera receives detection
Light intensity signal be converted to electric signal through NI data collecting cards 23 be sent to computer 24 can also carry out analysis obtain position with
The relationship of voltage, and then obtain the stiffness coefficient of ligh trap residing for sample to be tested and obtain sample to be tested in different viscoelastic bodies
Viscosity in system, wherein visible light and infrared interference filter may be used in high pass low-resistance optical filter 21.
In a preferred embodiment, the femtosecond pulse of high repetition frequency is since the nonlinear effect of itself can be with
It realizes two-photon excitation, two-photon excitation, therefore the excitation wavelength of selected fluorescent dye can be obtained again while capture
Appropriate, excitation light source unit 3 of the invention is using high stability TEM00The wavelength of mode continuous wave laser, continuous wave laser needs
To match with the dye molecule excited, excitation light source unit 3 includes that (the present embodiment uses 532nm waves to solid state laser 30
It is long, without being limited thereto as example), (exciting light bandpass filtering may be used in neutral density filter plate 31 to neutral density filter plate 31
Piece), several speculums 32, Glan-Taylor prism 33, quarter-wave plate 34, expand and collimating mirror 35, lens 36 and two filter
Mating plate 37；The continuous wave laser that solid state laser 30 is sent out is emitted to speculum 32a through neutral density filter plate 31, through speculum
The laser of 32a outgoing is emitted to through Glan-Taylor prism 33, quarter-wave plate 34 and speculum 32b successively to be expanded and collimates
Mirror 35, through expanding the circularly polarized light being emitted with collimating mirror through successively through lens 36, optical filter 37a, speculum 32c and optical filter
37b is focused on the back focal plane of microcobjective 40, since there is high x Microscope Objective 40 very high numerical aperture may be implemented entirely
Internal reflection can realize total internal reflection fluorescent imaging more higher than wide field fluorescence imaging signal-to-noise ratio.Wherein, the purpose expanded herein
Be for by excitation beam enlarged-diameter with ensure its size be more than microcobjective 40 light well to ensure that exciting light is high-quality
The circularly polarized light of amount.
In a preferred embodiment, EMCCD cameras may be used in single molecular fluorescence imaging unit 5 and pci data is adopted
Truck, EMCCD cameras connect computer 24 by pci data card, and EMCCD cameras have the high sensitivity of single photon, can be with
Obtain the out-of-focus image of single polymer molecule.
In a preferred embodiment, sample to be tested can be polystyrene fluorescence beads, gold nano grain, quantum
Point, non-blooming polystyrene sphere or silicon oxide pellets, Viable human cell；Wherein, polystyrene fluorescence beads radius
The radius of about 100nm, gold nano grain and quantum dot is 25nm, 30nm etc., much smaller than girdling the waist for femtosecond pulse laser 10
Radius, unstressed configuration polystyrene sphere or silicon oxide pellets radius are 0.80 μm, 1.08 μm, 2.02 μm etc., are close or larger than
The radius of the waist radius of femtosecond pulse laser 10, Viable human cell is 5 μm, is much larger than the beam of femtosecond pulse laser 10
Waist radius, therefore, the present invention can capture various sizes of Michaelis particle, obtain intuitive image.
Below by specific embodiment be described in detail it is using the present invention obtain in solution colloidal particle dynamics and at
As the use process of the femtosecond optical optical tweezers system of information.
Embodiment 1：The dynamic information of single colloidal particle in solution under different viscosities, specific mistake are obtained using the present invention
Cheng Wei：
1, the femtosecond optical optical tweezers system of the present invention is placed on optical table, adjusts each optics device in femtosecond optical optical tweezers system
The position of part makes the paths condition for meeting femtosecond optical optical tweezers system of the present invention；
2, it is that 1.08 μm of polystyrene microspheres are positioned over as sample to be tested in sample cell using radius, sample cell bottom is
The coverslip of 0.17mm；
3, the femtosecond pulse laser 10 of femtosecond capture laser light source unit 1 is opened so that femtosecond pulse laser 10 is sent out
The laser gone out is expanded to the aperture requirement that beam diameter meets microcobjective 40 through expanding with collimating mirror 12, and the laser after expanding is collimated
Make directional light；
4, microcobjective 40 is switched to oil immersion microcobjective, 20~100 μ L mirror oil is added on microcobjective 40, and right
It, which is adjusted, makes up to suitable focal position；
5, the halogen lamp of microscope 40 is opened, and adjusts and arrives suitable brightness, by digital camera or EMCCD cameras, just
Step finds the colloid bead to suspend in the solution；
6,23 gathered data of NI data collecting cards is opened；
7, data are exported after terminating data set, the statistical probability distribution figure (as shown in Figure 3) of out position is done, passes through
Boltzmann statistic laws calibrate to obtain Optical Trap Stiffness to be 0.800pN/ μm, and viscosity is 0.0013 pool (22 DEG C of indoor temperature).
8, the diffusion coefficient D that sample to be tested is calculated according to Stokes-einstein formula is 0.016 μm2/s。
Embodiment 2：The lower nano fluorescent bead fluorescence imaging of optical tweezer capture is completed using the present invention, detailed process is：
1, the femtosecond optical optical tweezers system of the present invention is placed on optical table, by the coated polystyrene sphere of fluorescent dye
It is placed in right over microcobjective 4, adjusts each optical device and make the paths condition for meeting the present invention,
2, sample to be tested is positioned in sample cell, sample cell bottom uses the coverslip of 0.17mm；
3, the solid state laser 30 for opening excitation light source unit 3 makes the continuous wave laser that solid state laser 30 is emitted through expanding
Exciting light is set to be focused at the back focal plane of microcobjective 40 after beam adjustment, so that being flat by being emitted after microcobjective 40
Row light；
4, microcobjective 40 is switched to oil immersion microcobjective, 20-100 μ L mirror oil is added on microcobjective 40, and right
It, which is adjusted, makes up to best focus position；
5, it opens 1 femtosecond pulse laser 10 in femtosecond capture laser light source unit and its light intensity is adjusted；
6, it opens single molecular fluorescence imaging unit 5 and real-time fluorescence imaging (as shown in Figure 4) is carried out to single fluorescence beads, lead to
Cross the fluorescence signal intensity stepped information for changing to obtain the colloidal particle variation number that is captured at any time.
The various embodiments described above are merely to illustrate the present invention, wherein the structure of each component, connection type and manufacture craft etc. are all
It can be varied from, every equivalents carried out based on the technical solution of the present invention and improvement should not exclude
Except protection scope of the present invention.
Claims (9)
1. a kind of obtaining the femtosecond optical optical tweezers system of colloidal particle dynamics and image-forming information in solution, it is characterised in that the system
Including：
Stablize the femtosecond optical tweezer capture laser cell for capturing ligh trap for capturing laser as femtosecond and being provided for sample to be tested；
Position sensing for detecting captured sample to be tested position distribution when doing limited Brownian movement relative to ligh trap center
Unit；
Excitation light source unit for carrying out fluorescence excitation as exciting light irradiation sample to be tested；
For femtosecond capture laser to be collected into the position detection unit, and the exciting light that the excitation light source unit is emitted
The sample to be tested being introduced into capture ligh trap so that the luminescent dye molecule of sample to be tested, which is stimulated, generates fluorescence signal
Optical microphotograph unit；
Fluorescence signal for collecting sample to be tested generation completes the real-time fluorescence of single colloidal particle coated to fluorescent dye
Imaging obtains the single molecular fluorescence of the dynamic information of single colloidal particle of the sample to be tested in different viscoelasticity characteristic systems
Imaging unit.
2. femtosecond optical optical tweezers system according to claim 1, which is characterized in that the optical microphotograph unit is using inversion fluorescence
Microscopic structure, including microcobjective, first~third dichroscope and condenser；Exciting light is focused on through the microcobjective
Sample to be tested in capture ligh trap, the fluorescence generated through excitation sample to be tested are sent out after microcobjective collection
It is mapped to the first dichroscope, the fluorescence signal being emitted through first dichroscope is emitted to the single molecular fluorescence imaging measurement
Unit.
3. femtosecond optical optical tweezers system according to claim 2, which is characterized in that the femtosecond optical tweezer captures laser cell and includes
Femtosecond pulse laser expands and collimating mirror and deflecting mirror, the femtosecond pulse warp of the femtosecond pulse laser outgoing
Described expand is expanded and is collimated with collimating mirror, and the femtosecond pulse after expanding and collimating is through the deflecting mirror into horizontal deflection
After be emitted to the second dichroscope, the light emitting through second dichroscope outgoing is to the microcobjective.
4. femtosecond optical optical tweezers system according to claim 2, which is characterized in that the position detection unit includes assembling thoroughly
Mirror, positional detecting device, data collecting card and computer；The interference pattern that forward scattering light through microcobjective outgoing is formed
Sample is emitted to third dichroscope after condenser collection, and the light through third dichroscope outgoing is assembled thoroughly through described
Mirror focuses the positional detecting device, and the positional detecting device connects the computer by the data collecting card and waited for
Viscosity of the stiffness coefficient and sample to be tested of ligh trap residing for sample in different viscoelastic systems.
5. femtosecond optical optical tweezers system according to claim 4, which is characterized in that the positional detecting device uses position sensing
Detector or digital camera.
6. femtosecond optical optical tweezers system according to claim 2, which is characterized in that the excitation light source unit includes Solid State Laser
Device, quarter-wave plate, expands and collimating mirror, lens and optical filter Glan-Taylor prism；What the solid state laser was sent out
Continuous wave successively through the Glan-Taylor prism and quarter-wave plate be emitted to it is described expand and collimating mirror, expanded through described
Circularly polarized light with collimating mirror outgoing through the upper lens and optical filter through focusing on the back focal plane of the microcobjective successively.
7. femtosecond optical optical tweezers system according to claim 6, which is characterized in that the solid state laser exit setting is neutral
Density filtering piece.
8. femtosecond optical optical tweezers system according to claim 4, which is characterized in that the single molecular fluorescence imaging unit uses
EMCCD cameras and pci data capture card, the EMCCD cameras connect the computer through the pci data capture card.
9. femtosecond optical optical tweezers system according to claim 1, which is characterized in that the sample to be tested is that polystyrene fluorescence is small
Ball, gold nano grain, non-blooming polystyrene sphere or silicon oxide pellets, Viable human cell.
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| CN110664369A (en) * | 2019-09-19 | 2020-01-10 | 哈尔滨工业大学 | Self-adaptive confocal line scanning harmonic microscopic imaging method and device |
| CN111366510A (en) * | 2020-03-02 | 2020-07-03 | 清华大学深圳国际研究生院 | Suspended particulate matter flux measuring device utilizing synchronous polarization and fluorescence |
| CN111965050A (en) * | 2020-08-19 | 2020-11-20 | 天津大学 | Method for testing shearing force of colloidal particle microfluid |
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| CN114755200B (en) * | 2022-03-21 | 2022-11-08 | 北京大学长三角光电科学研究院 | Visual monitoring system and method based on photodynamic therapy |
| CN115112632A (en) * | 2022-07-11 | 2022-09-27 | 浙江大学 | Aerosol chemical reaction device based on double-camera multi-optical-trap optical tweezers |
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