CN105699344A - Measuring system for obtaining single molecular spectrum and imaging under shear field - Google Patents
Measuring system for obtaining single molecular spectrum and imaging under shear field Download PDFInfo
- Publication number
- CN105699344A CN105699344A CN201610031083.XA CN201610031083A CN105699344A CN 105699344 A CN105699344 A CN 105699344A CN 201610031083 A CN201610031083 A CN 201610031083A CN 105699344 A CN105699344 A CN 105699344A
- Authority
- CN
- China
- Prior art keywords
- fluorescence
- unit
- single molecular
- light
- spectrum
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6402—Atomic fluorescence; Laser induced fluorescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N2021/6417—Spectrofluorimetric devices
- G01N2021/6419—Excitation at two or more wavelengths
Landscapes
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Optics & Photonics (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention relates to a measuring system for obtaining a single molecular spectrum and imaging under a shear field.The measuring system is characterized by comprising an excitation light source unit serving as excitation light to irradiate a sample to be measured, a shear application and rheological measurement unit used for applying mechanical shear to the sample to be measured and measuring the macro-rheology property of the sample to be measured at the same time, and an optical microscope unit used for leading excitation light emitted by the excitation light source unit into the sample to be measured under the shear field to excite dye molecules of the sample to be measured to generated fluorescence signals, collecting the generated fluorescence signals and sending the fluorescence signals to a single molecular fluorescence imaging unit, a single molecular fluorescence emission spectrum measuring unit and/or a fluorescence correlation spectrum measuring unit respectively.The measuring system can be widely applied to polyelectrolyte dilute solutions, gel, interface diffusion, thin film imaging and other multi-charge condensed state systems for exploring the basic physical root of special features of the multi-charge condensed state systems.
Description
Technical field
The invention belongs to Polymer Physics basic research field, obtain single molecular spectrum and imaging measurement system under shearing field especially with regard to one。
Background technology
The physics root of multi-charge soft substance system rheological property is always up the important basic research work in this field。The research of this respect not only will help us to design new function material, as: water treated substance, intellectual drug, tissue engineering material etc., moreover it is possible to help us understand basic life process。But, owing to this kind of soft substance system self is with multiple electric charges, the existence of long-range electrostatic interaction and a large amount of counter ion counterionsl gegenions, the rheological property of system is very special, and a lot of problems are not realized always。The instrument of the soft substance system macrorheology character such as synchro measure macromolecule and its microstructure characteristic parameter can being always up international academic community, the pursuing a goal of instrument manufacturing industry, therefore, people once carried out some good tries in this respect。Owing to scattering method is widely used in the sign of macromolecular solution, it is idea naturally by the two combination, therefore, small-angle neutron scattering and rheology measurement technology are combined by Yoshiaki etc., Chinese Academy of Sciences Han Zhi is superfine to combine laser light scattering with mechanical shearing technology, laser light scattering techniques is measured technology with capillary rheology and is combined by Qingdao University of Science and Technology Hu Haiqing etc., the scattering flow graph that Anton Paar company of the U.S. produces is by light scattering technique, X ray scattering technology and optical microscopy combine with stream measurement technology, above-mentioned technology all achieves while implementing dynamic rheological property measurement, structural information by the strand in the method for static light scattering acquisition polymeric fluid；Confocal fluorescent microscope is combined with rheology measurement and can observe under stream change shearing field by professor DanielL.Blair of Georgetown university of the U.S., and the macrostructure of fluid system develops；Particle trajectory tracer technique is combined with rheology measurement and has successfully carried out in the non-linear rheology measurement of polymer melt by professor Shi-QingWang of Akron university of the U.S., the observation of fluid macrostructure under shearing field。
But, these methods all cannot associating from the studied system microstructure information of monomolecular angle research and its macrorheology character, especially light scattering technique is subject to the restriction of solution concentration and scattering section, be not suitable for the structure measurement of multi-charge system, be unable to meet the needs of multi-charge architectural study。
Summary of the invention
For the problems referred to above, it is an object of the invention to provide a kind of measurement system obtaining single molecular spectrum and imaging under shearing field, it is capable of multi-charge system is carried out that accurately dynamic shearing can also carry out high-resolution while measuring, highly sensitive unimolecule is measured and imaging, and then obtains the rheological properties of multi-charge system macroscopic view and its microstructure information。
For achieving the above object, the present invention takes techniques below scheme: single molecular spectrum and imaging measurement system under a kind of acquisition shearing field, it is characterised in that this measurement system includes: one for the excitation source unit as excitation light irradiation testing sample；One for measuring shearing applying and the flow measurement unit of its macrorheology characteristic simultaneously to testing sample applying mechanical shearing, and described shearing applying and flow measurement unit bottom arrange an exciting light optical window；And an optical microphotograph unit, described optical microphotograph unit is for being incorporated into, through described exciting light optical window, the testing sample being positioned under shearing field by the exciting light of described excitation source unit outgoing, the dye molecule making testing sample is stimulated generation fluorescence signal, and the fluorescence signal collection of generation is transmitted into a single molecular fluorescence image-generating unit, a single molecular fluorescence emission spectroscopy measurements unit and/or a fluorescence correlation spectroscopy measuring unit respectively；Described single molecular fluorescence image-generating unit is for carrying out real-time fluorescence imaging to the single fluorescence molecule spread in slower system, it is thus achieved that the unimolecule real space movable information in the system that diffusion is slower；Described single molecular fluorescence emission spectroscopy measurements unit is for emission spectrum to fluorescence molecule single in system to be measured, and it is carried out intensity analysis obtains, by the change of spectrum, the information that the counter ion counterionsl gegenions near dye molecule are distributed；Described fluorescence correlation spectroscopy measuring unit is for obtaining the association spectrum of fluorescence signal, and its being associated property analysis obtains diffusion and the dimension information of polymer molecule。
Preferably, described shearing applies to adopt a torque rheometer with flow measurement unit, and described torque rheometer infrabasal plate arranges described exciting light optical window。
Preferably, described excitation source unit adopts continuous laser or femtosecond pulse, and the wavelength of described continuous laser or femtosecond pulse needs to match with the dye molecule excited；Described excitation source unit includes the laser instrument of more than one different wave length, some reflecting mirrors, some diaphragms, the first beam expanding lens, the second beam expanding lens, the incidence end of described second beam expanding lens arranges a neutral-density filter, and the light that described laser instrument sends is respectively through some described reflecting mirrors or diaphragm and carry out two-stage through described first beam expanding lens and the second beam expanding lens successively and expand excitation beam enlarged-diameter to guarantee that it is sized larger than the microscope objective light well of described optical microphotograph unit。
Preferably, described optical microphotograph unit adopts inverted fluorescence microscope structure, including a microscope objective, a dichroic mirror, the first～the 3rd reflecting mirror, a transmitting light high-pass filtering sheet, an exciting light band pass filter, a pin hole, a beam splitter, a slit and first～tertiary focusing lens；Described exciting light is transmitted into through described microcobjective and is positioned at the described testing sample sheared and apply with within flow measurement unit, the fluorescence excited through testing sample is transmitted into described dichroic mirror after described microcobjective is collected, it is transmitted into described first reflecting mirror through described transmitting light high-pass filtering sheet through the fluorescence signal of described dichroic mirror outgoing, the fluorescence reflected through described first reflecting mirror is transmitted into described single molecular fluorescence emission spectroscopy measurements unit through described first condenser lens and slit, it is transmitted into described second reflecting mirror through described second condenser lens through the fluorescence of described first reflecting mirror transmission, the light reflected through described second reflecting mirror is sent to described single molecular fluorescence image-generating unit, it is transmitted into described beam splitter through pin hole through the light of described second reflecting mirror transmission, it is transmitted into described fluorescence correlation spectroscopy measuring unit through described tertiary focusing lens through a part of light of described beam splitter outgoing；It addition, the exit end of described excitation source unit arranges described exciting light band pass filter。
Preferably, this measurement system also includes one second fluorescence correlation spectroscopy measuring unit, the 4th reflecting mirror and the 4th condenser lens, described 4th condenser lens is reflexed to through described 4th reflecting mirror, through the light emission of described 4th condenser lens outgoing to described second fluorescence correlation spectroscopy measuring unit through another part light of described beam splitter outgoing。
Preferably, each described fluorescence correlation spectroscopy measure Dan Jun include a single-photon detector with a data collecting card, the fluorescence signal that detection receives is converted to the signal of telecommunication and is sent to a computer through described data collecting card by described single-photon detector。
Preferably, described testing sample is polymer molecule, and described dye molecule is to receive on polymer molecular chain in the way of chemical bonding, described polymer molecule concentration is 10-9M。
Due to the fact that and take above technical scheme, it has the advantage that 1, the shearing applying of macroscopic view is combined by the present invention with flow measurement, single molecular fluorescence emission spectroscopy measurements, single molecular fluorescence imaging and fluorescence correlation spectroscopy measurement, the single molecular fluorescence emission spectrum of measured system and fluorescence correlation spectroscopy under acquisition shearing field；Additionally, system is carried out single molecular fluorescence imaging simultaneously, obtain the unimolecule real space movable information in spreading slower system, thus realizing that multi-charge system is carried out accurately dynamic shearing can also carry out high-resolution, the measurement of highly sensitive unimolecule and imaging while measuring, and then obtain the rheological properties of multi-charge system macroscopic view and the structural information of its microcosmic。2, the present invention is by offering exciting light optical window at flow graph infrabasal plate place, while macrorheology measurement can be realized, by this optical window, testing sample is carried out the collection of excitation and fluorescence signal, and then realize the measurement of unimolecule spectrum and imaging, finally give the microphysics image of testing sample macroscopic properties。3, the testing sample of the present invention is polymer molecule, and dye molecule is to receive on polymer molecular chain in the way of chemical bonding, polymer molecule concentration is 10-9M, therefore the present invention to extremely rare solution measurement, can finally realize the measurement to individual molecule。Present invention can be extensively applied to the multi-charge condensed state systems such as polyelectrolyte weak solution, gel, interface diffusion, thin film imaging, explore the basic physics root of its special nature。
Accompanying drawing explanation
Fig. 1 is the measuring principle schematic diagram of the present invention；
Fig. 2 is the flow graph lower substrate structure schematic diagram that the present invention offers optical window；
Fig. 3 is the excitation source unit light line structure schematic diagram of the present invention。
Detailed description of the invention
Below in conjunction with accompanying drawing, the present invention is carried out detailed description。It should be appreciated, however, that being provided only of accompanying drawing is more fully understood that the present invention, they should not be interpreted as limitation of the present invention。
As shown in Figure 1 and Figure 2, single molecular spectrum and imaging measurement system under the acquisition shearing field of the present invention, shear including an excitation source unit 1, and apply and flow measurement unit 2, optical microphotograph unit 3, single molecular fluorescence image-generating unit 4 and/or a spectral measurement unit, spectral measurement unit can adopt single molecular fluorescence emission spectroscopy measurements unit 5 and/or a fluorescence correlation spectroscopy measuring unit 6, wherein, shear bottom applying and flow measurement unit 2 and an exciting light optical window 21 is set；
The laser that excitation source unit 1 sends is as excitation light irradiation testing sample, and testing sample is polymer molecule, and dye molecule is to receive on polymer molecular chain in the way of chemical bonding；Shear and apply accurately to measure its macrorheology characteristic with flow measurement unit 2 for testing sample applies mechanical shearing simultaneously；Optical microphotograph unit 3 is for being incorporated into, by exciting light optical window 21, the testing sample being positioned at shearing flow after the match by the exciting light of LASER Light Source unit 1 outgoing, the dye molecule making testing sample is stimulated generation fluorescence signal, and the fluorescence signal collection of generation is transmitted into single molecular fluorescence image-generating unit 4, single molecular fluorescence emission spectroscopy measurements unit 5 and/or fluorescence correlation spectroscopy measuring unit 6 respectively；Single molecular fluorescence image-generating unit 4 is for carrying out real-time fluorescence imaging to the single fluorescence molecule spread in slower system (gel, macromolecular solid body thin film etc.), obtain the unimolecule real space movable information in spreading slower system, namely obtain diffusion and the orientation information of this fluorescence molecule；Single molecular fluorescence emission spectroscopy measurements unit 5 is for the emission spectrum to fluorescence molecule single in system to be measured, and it is carried out intensity analysis obtains, by the change of spectrum, the information that the counter ion counterionsl gegenions near dye molecule are distributed；Fluorescence correlation spectroscopy measuring unit 6 is for obtaining the association spectrum of fluorescence signal, and its being associated property analysis obtains diffusion and the dimension information of polymer molecule。
In a preferred embodiment, as shown in Figure 2, shear and apply to adopt, with flow measurement unit 2, the torque rheometer that accurate dynamic shearing can be implemented, torque rheometer infrabasal plate is provided with the exciting light optical window 21 of a high permeability and short operating distance, short operating distance is in order to the microscope water immersion objective with high-NA matches, its operating distance very short, maximum 0.2mm。
In a preferred embodiment, excitation source unit 1 can adopt continuous laser or femtosecond pulse, purpose is to excite more kinds of fluorescence molecule, the wavelength of selected laser instrument needs to match with the dye molecule excited, excitation source unit 1 in the present embodiment includes the laser instrument 10 of three kinds of different wave lengths (as example, but it is not limited to this, can select according to actually used), some reflecting mirrors 11, some diaphragms 12, first beam expanding lens 13 and the second beam expanding lens 14, the incidence end of the second beam expanding lens 14 can arrange one for regulating the neutral-density filter 15 of the power of exciting light, the wavelength of three laser instrument 10 respectively 1060nm, 532nm and 473nm, three laser instrument 10 reflect or transmission and diaphragm 12 carry out two-stage through the first beam expanding lens 13 and the second beam expanding lens 14 successively and expand excitation beam enlarged-diameter to about 2.0cm to guarantee that it is sized larger than the microscope objective light well of optical microphotograph unit 3 through some reflecting mirrors 11 respectively successively, make full use of the numerical aperture of object lens thus obtaining and produce minimum to excite space。
In a preferred embodiment, as shown in Figure 1, optical microphotograph unit 3 can adopt inverted fluorescence microscope structure, launches light high-pass filtering sheet 34, pin hole 35, beam splitter 36, slit 37, first～tertiary focusing lens 38 and an exciting light band pass filter 39 including microscope objective 31, dichroic mirror the 32, first～the 3rd reflecting mirror 33,；Exciting light is transmitted into be positioned at through microcobjective 31 shears the testing sample applied with within flow measurement unit 3, the fluorescence excited through testing sample is transmitted into dichroic mirror 32 after microcobjective 35 is collected, it is transmitted into the first reflecting mirror 33 through the emitted smooth high-pass filtering sheet 34 of fluorescence signal of dichroic mirror 32 outgoing, the fluorescence reflected through the first reflecting mirror 33 is transmitted into single molecular fluorescence emission spectroscopy measurements unit 5 through the first condenser lens 38 and slit 37, it is transmitted into the second reflecting mirror 33 through the second condenser lens 38 through the fluorescence of the first reflecting mirror 33 transmission, the light reflected through the second reflecting mirror 33 is sent to single molecular fluorescence image-generating unit 4, it is transmitted into beam splitter 36 through pin hole 35 through the light of the second reflecting mirror 33 transmission, it is transmitted into fluorescence correlation spectroscopy measuring unit 6 through tertiary focusing lens 38 through a part of light of beam splitter 36 outgoing；It addition, the exit end of excitation source unit 1 arranges exciting light band pass filter 39。
The glitch that after-pulsing in order to effectively eliminate detector itself produces, present invention additionally comprises one second fluorescence correlation spectroscopy measuring unit the 6, the 4th reflecting mirror 33 and the 4th condenser lens 38, the 4th condenser lens 38 is reflexed to through the 4th reflecting mirror 33, through light emission the second fluorescence correlation spectroscopy measuring unit 6 of the 4th condenser lens 38 outgoing through another part light of beam splitter 36 outgoing。
In a preferred embodiment, single molecular fluorescence image-generating unit 4 can adopt EMCCD camera。
In a preferred embodiment, single molecular fluorescence emission spectroscopy measurements unit 5 can adopt spectroanalysis instrument。
In a preferred embodiment, each fluorescence correlation spectroscopy measuring unit 6 all include a single-photon detector with a data collecting card, the fluorescence signal that detection receives is converted to the signal of telecommunication and is sent to computer being associated property analysis acquisition correlation function data through data collecting card by single-photon detector, and then obtains the diffusion coefficient of testing sample, size, directed movement speed and mean concentration information。
In a preferred embodiment, present invention additionally comprises a high precision displacement platform, torque rheometer can being placed on the top of microscope objective 31, thus realizing the accurate linkage of each device during use, high precision displacement platform is the displacement platform that can adopt prior art, does not repeat them here。
In a preferred embodiment, the concentration of polymer molecule can be 10-9M。
The measurement process adopting the present invention to obtain single molecular spectrum and imaging measurement system under shearing field is described in detail below by specific embodiment。
Embodiment 1: obtaining shearing field and place an order molecular fluorescence spectroscopy and fluorescence correlation spectroscopy, detailed process is:
1, the present invention is placed on an optical table, flow graph is moved to directly over microscope objective 31 by high precision position moving stage, adjust each optics and make to meet the optic path condition of the present invention,
2, testing sample is positioned over the microscope slide center being positioned at flow graph, and microscope slide adopts thickness to be the quartz slide of 0.13～0.17mm；
3, open the laser instrument of 1 in excitation source unit so that the laser that laser instrument sends carries out two-stage through beam expanding lens and expands, and the laser after expanding is carried out collimation make directional light；
4, microscope objective 31 is switched to water immersion objective, microscope objective 31 adds 20-100 μ L ultra-pure water, and adjusted makes up to suitable focal position；
5, open flow graph and implement dynamic shearing and flow measurement；
6, adjustment pin hole 35 or slit 37 position make the fluorescence signal that fluorescence correlation spectroscopy measuring unit 6 or single molecular fluorescence emission spectroscopy measurements unit 5 detect best；
7, open the information that single molecular fluorescence emission spectroscopy measurements unit 5 obtains the fluorescence emission spectrum of individual molecule, and obtain the counter ion counterionsl gegenions distributed intelligence near dye molecule by analyzing the change of spectrum。
8, open fluorescence correlation spectroscopy measuring unit 6 and obtain the associated light spectral curve of fluorescence signal, and obtained diffusion and the dimension information of polymer molecule by data fitting。
Embodiment 2: complete the measurement of single molecular fluorescence imaging under shearing field, detailed process is:
1, the present invention is placed on an optical table, flow graph is moved to directly over microscope objective 31 by high precision position moving stage, adjust each optics and make to meet the optic path condition of the present invention,
2, testing sample is positioned over the microscope slide center being positioned at flow graph, and microscope slide adopts thickness to be the quartz slide of 0.13～0.17mm；
3, open the laser instrument of excitation source unit 1, make the laser of laser instrument 10 outgoing make exciting light focus at the negative focal plane place of microscope objective 31 after beam expanding lens adjustment, so that be directional light by outgoing after microscope objective 31；
4, microscope objective 31 is switched to oil immersion objective, microscope objective 31 adds 20-100 μ L mirror oil, and adjusted makes up to best focus position；
5, open flow graph and implement dynamic shearing and flow measurement；
6, open single molecular fluorescence image-generating unit 4 and single fluorescence molecule is carried out real-time fluorescence imaging, thus obtaining the diffusion of molecule and the information of orientation。
The various embodiments described above are merely to illustrate the present invention; wherein the structure of each parts, connected mode and processing technology etc. all can be varied from; every equivalents carried out on the basis of technical solution of the present invention and improvement, all should not get rid of outside protection scope of the present invention。
Claims (8)
1. one kind obtains single molecular spectrum and imaging measurement system under shearing field, it is characterised in that this measurement system includes:
One for the excitation source unit as excitation light irradiation testing sample；
One for measuring shearing applying and the flow measurement unit of its macrorheology characteristic simultaneously to testing sample applying mechanical shearing, and described shearing applying and flow measurement unit bottom arrange an exciting light optical window；And
One optical microphotograph unit, described optical microphotograph unit is for being incorporated into, through described exciting light optical window, the testing sample being positioned under shearing field by the exciting light of described excitation source unit outgoing, the dye molecule making testing sample is stimulated generation fluorescence signal, and the fluorescence signal collection of generation is transmitted into a single molecular fluorescence image-generating unit, a single molecular fluorescence emission spectroscopy measurements unit and/or a fluorescence correlation spectroscopy measuring unit respectively；Described single molecular fluorescence image-generating unit is for carrying out real-time fluorescence imaging to the single fluorescence molecule spread in slower system, it is thus achieved that the unimolecule real space movable information in the system that diffusion is slower；Described single molecular fluorescence emission spectroscopy measurements unit is for emission spectrum to fluorescence molecule single in system to be measured, and it is carried out intensity analysis obtains, by the change of spectrum, the information that the counter ion counterionsl gegenions near dye molecule are distributed；Described fluorescence correlation spectroscopy measuring unit is for obtaining the association spectrum of fluorescence signal, and its being associated property analysis obtains diffusion and the dimension information of polymer molecule。
2. single molecular spectrum and imaging measurement system under a kind of acquisition shearing field as claimed in claim 1, it is characterised in that described shearing applying and flow measurement unit adopt a torque rheometer, and described torque rheometer infrabasal plate arranges described exciting light optical window。
3. single molecular spectrum and imaging measurement system under a kind of acquisition shearing field as claimed in claim 1, it is characterized in that, described excitation source unit adopts continuous laser or femtosecond pulse, and the wavelength of described continuous laser or femtosecond pulse needs to match with the dye molecule excited；Described excitation source unit includes the laser instrument of more than one different wave length, some reflecting mirrors, some diaphragms, the first beam expanding lens, the second beam expanding lens, the incidence end of described second beam expanding lens arranges a neutral-density filter, and the light that described laser instrument sends is respectively through some described reflecting mirrors or diaphragm and carry out two-stage through described first beam expanding lens and the second beam expanding lens successively and expand excitation beam enlarged-diameter to guarantee that it is sized larger than the microscope objective light well of described optical microphotograph unit。
4. single molecular spectrum and imaging measurement system under a kind of acquisition shearing field as claimed in claim 2, it is characterized in that, described excitation source unit adopts continuous laser or femtosecond pulse, and the wavelength of described continuous laser or femtosecond pulse needs to match with the dye molecule excited；Described excitation source unit includes the laser instrument of more than one different wave length, some reflecting mirrors, some diaphragms, the first beam expanding lens, the second beam expanding lens, the incidence end of described second beam expanding lens arranges a neutral-density filter, and the light that described laser instrument sends is respectively through some described reflecting mirrors or diaphragm and carry out two-stage through described first beam expanding lens and the second beam expanding lens successively and expand excitation beam enlarged-diameter to guarantee that it is sized larger than the microscope objective light well of described optical microphotograph unit。
5. one as claimed in claim 1 or 2 or 3 or 4 obtains single molecular spectrum and imaging measurement system under shearing field, it is characterized in that, described optical microphotograph unit adopts inverted fluorescence microscope structure, including a microscope objective, a dichroic mirror, the first～the 3rd reflecting mirror, a transmitting light high-pass filtering sheet, an exciting light band pass filter, a pin hole, a beam splitter, a slit and first～tertiary focusing lens；Described exciting light is transmitted into through described microcobjective and is positioned at the described testing sample sheared and apply with within flow measurement unit, the fluorescence excited through testing sample is transmitted into described dichroic mirror after described microcobjective is collected, it is transmitted into described first reflecting mirror through described transmitting light high-pass filtering sheet through the fluorescence signal of described dichroic mirror outgoing, the fluorescence reflected through described first reflecting mirror is transmitted into described single molecular fluorescence emission spectroscopy measurements unit through described first condenser lens and slit, it is transmitted into described second reflecting mirror through described second condenser lens through the fluorescence of described first reflecting mirror transmission, the light reflected through described second reflecting mirror is sent to described single molecular fluorescence image-generating unit, it is transmitted into described beam splitter through pin hole through the light of described second reflecting mirror transmission, it is transmitted into described fluorescence correlation spectroscopy measuring unit through described tertiary focusing lens through a part of light of described beam splitter outgoing；It addition, the exit end of described excitation source unit arranges described exciting light band pass filter。
6. single molecular spectrum and imaging measurement system under a kind of acquisition shearing field as claimed in claim 5, it is characterized in that, this measurement system also includes the second fluorescence correlation spectroscopy measuring unit, the 4th reflecting mirror and the 4th condenser lens, described 4th condenser lens is reflexed to through described 4th reflecting mirror, through the light emission of described 4th condenser lens outgoing to described second fluorescence correlation spectroscopy measuring unit through another part light of described beam splitter outgoing。
7. the one as described in claim 1 or 2 or 3 or 6 obtains single molecular spectrum and imaging measurement system under shearing field, it is characterized in that, each described fluorescence correlation spectroscopy measure Dan Jun include a single-photon detector with a data collecting card, the fluorescence signal that detection receives is converted to the signal of telecommunication and is sent to a computer through described data collecting card by described single-photon detector。
8. the one as described in claim 1 or 2 or 3 or 6 obtains single molecular spectrum and imaging measurement system under shearing field, it is characterized in that, described testing sample is polymer molecule, and described dye molecule is to receive on polymer molecular chain in the way of chemical bonding, described polymer molecule concentration is 10-9M。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610031083.XA CN105699344A (en) | 2016-01-18 | 2016-01-18 | Measuring system for obtaining single molecular spectrum and imaging under shear field |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610031083.XA CN105699344A (en) | 2016-01-18 | 2016-01-18 | Measuring system for obtaining single molecular spectrum and imaging under shear field |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105699344A true CN105699344A (en) | 2016-06-22 |
Family
ID=56226560
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610031083.XA Pending CN105699344A (en) | 2016-01-18 | 2016-01-18 | Measuring system for obtaining single molecular spectrum and imaging under shear field |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105699344A (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107518879A (en) * | 2017-10-11 | 2017-12-29 | 北京数字精准医疗科技有限公司 | A kind of fluoroscopic imaging device and method |
| CN107744382A (en) * | 2017-11-20 | 2018-03-02 | 北京数字精准医疗科技有限公司 | Optical molecular image navigation system |
| CN108072636A (en) * | 2016-11-14 | 2018-05-25 | 中国科学院化学研究所 | A kind of single annular polymeric molecule spreads the measuring method of information in solid/liquid interfaces |
| CN108333151A (en) * | 2017-01-19 | 2018-07-27 | 北京大学 | A kind of super-resolution micro imaging system and imaging method based on femtosecond pulse shaping |
| CN108535229A (en) * | 2018-04-12 | 2018-09-14 | 中国科学院化学研究所 | A kind of obtain is limited the measurement method of single polymer molecule diffusion and motor pattern in melt |
| CN108550128A (en) * | 2018-04-20 | 2018-09-18 | 中国科学院化学研究所 | A kind of single molecular fluorescence out-of-focus image processing method |
| CN108760771A (en) * | 2018-07-24 | 2018-11-06 | 电子科技大学 | A method of tracer being carried out to drug using near-field microwave microscopy |
| CN110057751A (en) * | 2018-01-19 | 2019-07-26 | 清华大学 | The making apparatus and method of optical particle probe |
| CN110231321A (en) * | 2019-06-25 | 2019-09-13 | 中国科学院长春应用化学研究所 | Super-resolution micro imaging system is scanned based on nano-pore-lenticule |
| CN111337469A (en) * | 2020-04-27 | 2020-06-26 | 中国科学院化学研究所 | Method for acquiring microscopic motion information under shear field by utilizing trace light intensity distribution of fluorescent wide-field imaging |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030008296A1 (en) * | 2000-03-27 | 2003-01-09 | Olympus Optical Co., Ltd. | Method for determining polymorphism |
| CN101019018A (en) * | 2004-09-06 | 2007-08-15 | 浜松光子学株式会社 | Fluorescent microscope and fluorescent correlation spectral analysis device |
| CN101718696A (en) * | 2009-12-10 | 2010-06-02 | 上海交通大学 | Lasing fluorescence scanning imaging-fluorescence correlation spectrum unimolecule detecting instrument |
| US20120252056A1 (en) * | 2009-10-13 | 2012-10-04 | Universite De Technologies De Troyes | Imaging method and system using substrate functionalization |
-
2016
- 2016-01-18 CN CN201610031083.XA patent/CN105699344A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030008296A1 (en) * | 2000-03-27 | 2003-01-09 | Olympus Optical Co., Ltd. | Method for determining polymorphism |
| CN101019018A (en) * | 2004-09-06 | 2007-08-15 | 浜松光子学株式会社 | Fluorescent microscope and fluorescent correlation spectral analysis device |
| US20120252056A1 (en) * | 2009-10-13 | 2012-10-04 | Universite De Technologies De Troyes | Imaging method and system using substrate functionalization |
| CN101718696A (en) * | 2009-12-10 | 2010-06-02 | 上海交通大学 | Lasing fluorescence scanning imaging-fluorescence correlation spectrum unimolecule detecting instrument |
Non-Patent Citations (4)
| Title |
|---|
| CHRISTINE M. PAPADAKIS 等: "Polymers in focus: fluorescence correlation spectroscopy", 《COLLOID POLYM SCI》 * |
| SILKE RATHGEBER 等: "Microrheology with Fluorescence Correlation Spectroscopy", 《LANGMUIR》 * |
| 王生勤 等: "单分子荧光技术与聚合物单链物理性质的研究", 《高分子通报》 * |
| 郑锴锴 等: "获取剪切场下单个高分子的光谱信息-高分子流变显微光谱仪", 《2015年全国高分子学术论文报告会论文摘要集——主题C 高分子物理与软物质》 * |
Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108072636A (en) * | 2016-11-14 | 2018-05-25 | 中国科学院化学研究所 | A kind of single annular polymeric molecule spreads the measuring method of information in solid/liquid interfaces |
| CN108333151A (en) * | 2017-01-19 | 2018-07-27 | 北京大学 | A kind of super-resolution micro imaging system and imaging method based on femtosecond pulse shaping |
| CN108333151B (en) * | 2017-01-19 | 2021-01-26 | 北京大学 | Super-resolution microscopic imaging system and method based on femtosecond pulse shaping |
| CN107518879A (en) * | 2017-10-11 | 2017-12-29 | 北京数字精准医疗科技有限公司 | A kind of fluoroscopic imaging device and method |
| CN107744382A (en) * | 2017-11-20 | 2018-03-02 | 北京数字精准医疗科技有限公司 | Optical molecular image navigation system |
| CN110057751A (en) * | 2018-01-19 | 2019-07-26 | 清华大学 | The making apparatus and method of optical particle probe |
| CN110057751B (en) * | 2018-01-19 | 2021-06-29 | 清华大学 | Apparatus and method for fabricating optical particle probe |
| CN108535229B (en) * | 2018-04-12 | 2020-10-27 | 中国科学院化学研究所 | Measurement method for obtaining diffusion and movement modes of single polymer molecules in limited melt |
| CN108535229A (en) * | 2018-04-12 | 2018-09-14 | 中国科学院化学研究所 | A kind of obtain is limited the measurement method of single polymer molecule diffusion and motor pattern in melt |
| CN108550128B (en) * | 2018-04-20 | 2020-08-04 | 中国科学院化学研究所 | Single-molecule fluorescence defocusing image processing method |
| CN108550128A (en) * | 2018-04-20 | 2018-09-18 | 中国科学院化学研究所 | A kind of single molecular fluorescence out-of-focus image processing method |
| CN108760771A (en) * | 2018-07-24 | 2018-11-06 | 电子科技大学 | A method of tracer being carried out to drug using near-field microwave microscopy |
| CN108760771B (en) * | 2018-07-24 | 2022-10-14 | 电子科技大学 | Method for tracing medicine by using near-field microwave microscope |
| CN110231321A (en) * | 2019-06-25 | 2019-09-13 | 中国科学院长春应用化学研究所 | Super-resolution micro imaging system is scanned based on nano-pore-lenticule |
| CN110231321B (en) * | 2019-06-25 | 2020-05-12 | 中国科学院长春应用化学研究所 | Super-resolution microscopic imaging system based on nanopore-microlens scanning |
| CN111337469A (en) * | 2020-04-27 | 2020-06-26 | 中国科学院化学研究所 | Method for acquiring microscopic motion information under shear field by utilizing trace light intensity distribution of fluorescent wide-field imaging |
| CN111337469B (en) * | 2020-04-27 | 2021-06-08 | 中国科学院化学研究所 | Method for acquiring microscopic motion information under shear field by utilizing fluorescence wide field imaging |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105699344A (en) | Measuring system for obtaining single molecular spectrum and imaging under shear field | |
| US11698334B2 (en) | Flow cytometer with optical equalization | |
| CN105388140B (en) | Measuring instrument for site invisible fingerprint display and contained substance thereof | |
| CN103743718A (en) | Laser spectrum analyzer combining confocal micro-Raman spectrometer with laser-induced breakdown spectrometer | |
| JPS6394156A (en) | Method and instrument for analyzing cell in fluid | |
| US10539495B2 (en) | Systems and methods for generating an image of an inspection object using an attenuated beam | |
| CN104749162B (en) | Confocal laser-scanning microscopy instrument and its light path device | |
| JP2005534944A (en) | Fluorescence correlation spectrometer | |
| CN105675560B (en) | A kind of method of single polymer molecule fluorescence emission spectrum information under acquisition shearing field | |
| CN113008849B (en) | Ultraviolet-near infrared broadband micro-region photoluminescence spectrum testing device | |
| CN108801863A (en) | The femtosecond optical optical tweezers system of colloidal particle dynamics and image-forming information in solution can be obtained | |
| US10215630B2 (en) | Raman spectroscopy systems and raman spectroscopy methods | |
| JP2022532496A (en) | Phase calibration for imaging flow cytometry | |
| CN105675561A (en) | Method for acquiring diffusive and directional movement information of single polymer molecules in shear fields | |
| CN106885795A (en) | A kind of fluorescence lifetime information acquisition method and system for moving single-particle | |
| JP2005043278A (en) | Fluorescence correlation spectrometric apparatus and method of measuring diffusion coefficient for heterogeneous sample | |
| Świt et al. | Beam characterization of a microfading tester: evaluation of several methods | |
| Paiè et al. | Structured-light-sheet imaging in an integrated optofluidic platform | |
| EP1260810A2 (en) | Method for calibrating a sample analyzer | |
| JP5320593B2 (en) | Molecular analysis using laser-induced shock waves | |
| Dey | Fluorescence microscope, confocal microscope and other advanced microscopes: basic principles and applications in pathology | |
| TWI661222B (en) | Optical measuring device and optical measuring method | |
| Bennet | Multi-parameter quantitative mapping of microfluidic devices | |
| JP4016328B2 (en) | Viscosity measuring method and measuring apparatus | |
| JP2836859B2 (en) | Three-dimensional spatial and time-resolved absorption spectrum measurement device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20180205 Address after: 100190 Haidian District, Zhongguancun, North Street, No. 1, No. 2, Beijing Applicant after: Institute of Chemistry, Chinese Academy of Sciences Applicant after: University of Chinese Academy of Sciences Address before: 100190 Haidian District, Zhongguancun, North Street, No. 1, No. 2, Beijing Applicant before: Institute of Chemistry, Chinese Academy of Sciences |
|
| TA01 | Transfer of patent application right | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160622 |
|
| RJ01 | Rejection of invention patent application after publication |