CN204514812U - Confocal laser-scanning microscopy instrument and light path device thereof - Google Patents
Confocal laser-scanning microscopy instrument and light path device thereof Download PDFInfo
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- CN204514812U CN204514812U CN201520234553.3U CN201520234553U CN204514812U CN 204514812 U CN204514812 U CN 204514812U CN 201520234553 U CN201520234553 U CN 201520234553U CN 204514812 U CN204514812 U CN 204514812U
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Abstract
The utility model discloses Confocal laser-scanning microscopy instrument and light path device thereof, light path device is included in the following structure that light path sets gradually: excitation light source system, and described excitation light source system comprises excitation source and beam expanding lens; Raman filter set, Raman filter set described in described Raman filter set has side and receives glancing incidence light and the filter sheet structure forming the emergent light vertical with incident light at opposite side; Microscopic system, described microscopic system comprises the Guan Jing of the microcobjective being arranged at described Raman filter set sample emergent light side respectively and the sample emergent light offside being arranged at described Raman filter set; Monochromator, the Raman signal converged from described pipe mirror is incident upon described monochromator; And single photon counter, described single photon counter detection is from the Raman signal of described monochromator.According to the light path device of the Confocal laser-scanning microscopy instrument of the utility model embodiment, light path debug and simply, light path is shorter, sensing range is wider.
Description
Technical field
The utility model relates to optical instrument field, particularly a kind of Confocal laser-scanning microscopy instrument and light path device thereof.
Background technology
Raman spectrum is a kind of scattering spectrum, and it is that nineteen twenty-eight India physicist C handrasekhara Venkata Raman finds, and therefore obtains the Nobel Prize in physics of nineteen thirty.Raman spectrum as a kind of material molecular structure analytical test means and be widely used, especially after nineteen sixty First laser instrument appearance, the introducing of LASER Light Source, the raising of Technique of Weak Signal Detection and computer application, make Raman spectrum analysis obtain very large progress in many applications.At present, Raman spectrum has been widely used in the fields such as oil, food, agriculture and animal husbandry, macromolecule, pharmacy, biology, medical science.
Raman spectrometer principle of work is as follows: when a branch of frequency is ν
0monochromatic light when inciding solid, liquid or gaseous medium, the faint scattered light of to all the winds radiation can be caused.Stronger in scattered light is Rayleigh (Rayleigh) scattering, its frequency and incident light frequency ν
0identical, its intensity and the order of magnitude are about 10 of incident intensity
-4~ 10
-3.Except Rayleigh scattering, also have Raman scattering, the scattered light frequency ν of Raman scattering and incident light frequency have obvious change: ν=ν
0± | Δ ν |, its intensity is about 10 of Rayleigh scattering
-8~ 10
-6, be 10 of incident intensity
-12~ 10
-9.The frequency of Raman scattering line is by incident light frequency and the intrinsic internal motion frequency synthesis of molecule, so also referred to as associating scattered light.The frequency of Raman line changes with the change of incident light frequency, but remains unchanged relative to frequency-splitting (frequency displacement) the Δ ν of incident light frequency, and has nothing to do with the frequency of incident light, only relevant with the molecular structure of scattering medium itself.The vibration of molecule, rotation and spin determine different media to there being different Raman frequency shift spectrograms.
The light path device of existing Raman spectrometer exist debugging complicated, excite focal beam spot bigger than normal, the liquid such as water etc. that Raman signal is more weak is difficult to carry out the shortcoming such as measuring.
Utility model content
The utility model is intended to solve one of technical matters in correlation technique at least to a certain extent.For this reason, an object of the present utility model is to propose a kind ofly have the Confocal laser-scanning microscopy instrument and light path device thereof that light channel structure is simple, detection signal scope is wide.
According to the light path device of the Confocal laser-scanning microscopy instrument of the utility model embodiment, be included in the following structure that light path sets gradually: excitation light source system, described excitation light source system comprises excitation source and beam expanding lens, Raman filter set, described Raman filter set has side and receives glancing incidence light and the filter sheet structure forming the emergent light vertical with incident light at opposite side, microscopic system, described microscopic system comprises the Guan Jing of the microcobjective being arranged at described Raman filter set sample emergent light side respectively and the sample emergent light offside being arranged at described Raman filter set, monochromator, from monochromator described in the Raman signal directive that described pipe mirror converges, and single photon counter, described single photon counter detection is from the Raman signal of described monochromator, wherein, the laser sent from described excitation source is expanded by described beam expanding lens, through described Raman filter set to described microcobjective, and by described microcobjective, described Laser Focusing is inspired Raman signal on sample, described Raman signal becomes directional light and filters Rayleigh scattering by described Raman filter set again after described microcobjective, after this entrance slit place of described monochromator is focused on through described Guan Jing, and detected by the described single photon counter at exit slit place.
According to the light path device of the Confocal laser-scanning microscopy instrument of the utility model embodiment, all adopt vertical stratification at all turnover positions of light path, therefore light path debug and simply, light path is shorter, thus effectively can reduce the loss of Raman signal, be conducive to expanding sensing range; In addition, owing to adopting beam expanding lens to expand laser, the effective aperture of microcobjective can be used fully; Further, owing to using single photon counter as the detector collecting Raman signal, can by suitably increasing the intensity improving tested Raman signal integral time.
In addition, according to the light path device of the Confocal laser-scanning microscopy instrument of the utility model above-described embodiment, following additional technical characteristic can also be had:
According to embodiment of the present utility model, described Raman filter set comprises the vertical narrow band pass filter, two placed to the Rayleigh optical filter of light splitting part and horizontal positioned, described microcobjective is arranged at described two to light splitting part emergent light side, and described Guan Jing is arranged at the emergent light side of described Rayleigh optical filter.
According to embodiment of the present utility model, described two is 45 ° of plane mirrors to light splitting part.
According to embodiment of the present utility model, described light path device also comprises one or many reflection unit, and described reflection unit accepts the Raman signal from described pipe mirror convergence and reflexed to described monochromator.
According to embodiment of the present utility model, described reflection unit is plane mirror.
According to embodiment of the present utility model, described microcobjective adopts apochromatism flat field infinite distance metallography microscope object lens.
According to embodiment of the present utility model, the numerical aperture NA of described microcobjective can be more than 0.4.Owing to have employed the microcobjective of large-numerical aperture, be conducive to the collection efficiency increasing Raman scattering, even make launching efficiency only have 10
-6even lower Raman signal detection is achieved and commercialization.
According to embodiment of the present utility model, described light path device also comprises solid sample imaging system, described solid sample imaging system comprises push-and-pull reflection unit, CCD imaging device, described push-and-pull reflection unit is used for CCD target surface sample signal being imaged in described CC imaging device, to realize the focusing of solid sample, thus, focusing and the detection of solid sample can be realized, described push-and-pull reflection unit is with carriage, when solid sample need realize focusing, described push-and-pull reflection unit moves in light path, complete defocused, described push-and-pull reflection unit shifts out light path.
According to embodiment of the present utility model, described CCD target surface to described Guan Jing Distance geometry described in the entrance slit of monochromator equal to the distance of described Guan Jing.Easily can realize focusing and the detection of solid sample like this, realize cleverly focusing and the conversion detected by push-and-pull catoptron.
According to embodiment of the present utility model, the diameter focusing on hot spot is on to the sample less than 1.6 μm.Thus, laser energy can be enable more concentrated, thus greatly can improve the exciting light energy of Raman scattering.In one example, by beam expanding lens carry out more than 7 times expand the focusing realizing above-mentioned hot spot.
According to another aspect of the present utility model, also proposed the Confocal laser-scanning microscopy instrument with above-mentioned arbitrary light path device.
Accompanying drawing explanation
Fig. 1 is the structural representation of the light path device of Confocal laser-scanning microscopy instrument according to the utility model embodiment.
Fig. 2 is the structural representation according to the solid sample imaging system in the light path device of the Confocal laser-scanning microscopy instrument of the utility model embodiment.
In figure, 101 is excitation source, and 102 is beam expanding lens, 201 is Raman filter set, and 301 is microcobjective, and 302 is Guan Jing, 801 is monochromator, and 901 is single photon counter, and 401 is sample, 501 is plane mirror, 601 is push-and-pull plane mirror, and 701 is CCD, and 2011 is narrow band pass filter, 2012 is two to light splitting part, and 2013 is Rayleigh optical filter.
Embodiment
Be described below in detail embodiment of the present utility model, the example of described embodiment is shown in the drawings, and wherein same or similar label represents same or similar element or has element that is identical or similar functions from start to finish.Be exemplary below by the embodiment be described with reference to the drawings, be intended to for explaining the utility model, and can not be interpreted as restriction of the present utility model.
Below, be first described with reference to the light path device of accompanying drawing to the Confocal laser-scanning microscopy instrument of the utility model embodiment.
As shown in Figure 1, according to the light path device of the Confocal laser-scanning microscopy instrument of the utility model embodiment, be included in the excitation light source system (comprising 101 and 102) that light path sets gradually, described excitation light source system comprises excitation source 101 and beam expanding lens 102, Raman filter set 201, described Raman filter set 201 has side and receives glancing incidence light and the filter sheet structure forming the emergent light vertical with incident light at opposite side, microscopic system, described microscopic system comprises the pipe mirror 302 of the microcobjective 301 being arranged at described Raman filter set 201 sample emergent light side respectively and the sample emergent light offside being arranged at described Raman filter set 201, monochromator 801, from monochromator 801 described in the Raman signal directive that described pipe mirror 302 converges, and single photon counter 901, described single photon counter 901 detects the Raman signal from described monochromator 801, wherein, the laser sent from described excitation source 101 is expanded by described beam expanding lens 102, through described Raman filter set 201 to described microcobjective 301, and by described microcobjective 301, described Laser Focusing is inspired Raman signal on sample, described Raman signal becomes directional light and filters Rayleigh scattering by described Raman filter set 201 again after described microcobjective 301, after this entrance slit place of described monochromator 801 is focused on through described pipe mirror 302, and detected by the described single photon counter 901 at exit slit place.
As shown in Figure 2, described Raman filter set 201 comprises the Rayleigh optical filter 2013 of the vertical narrow band pass filter 2011, two placed to light splitting part 2012 and horizontal positioned, described microcobjective 301 is arranged at described two to light splitting part 2012 emergent light side, and described pipe mirror 302 is arranged at the emergent light side of described Rayleigh optical filter 2013.
Two to light splitting part 2012 be 45 ° two to light splitting part, more preferably, be 45 ° two to spectroscope.
Microscopic system comprise be arranged at respectively two to light splitting part 2012 emergent light side microcobjective 301 and be arranged at the pipe mirror 302 of emergent light side of Rayleigh optical filter 2013.
The Raman signal converged from pipe mirror 302 is incident upon monochromator 801.
Single photon counter 901 detects the Raman signal from monochromator 801.
In an embodiment of the present utility model, described light path device also comprises one or many reflection unit, and described reflection unit accepts the Raman signal from described pipe mirror convergence and reflexed to described monochromator.
More preferably, described reflection unit is plane mirror 501, alternatively, is 45 ° of plane mirrors.
In an embodiment of the present utility model, select the DPSS laser instrument of overstable, the minimum frequency displacement of 50mW wavelength 532nm as excitation source 101.
Wherein, microscopic system adopts apochromatism flat field infinite distance metallography microscope object lens respectively.
Wherein, the numerical aperture NA of microcobjective 301 can be more than 0.4.Owing to have employed the microcobjective of large-numerical aperture, be conducive to the collection efficiency increasing Raman scattering, even make launching efficiency only have 10
-6even lower Raman signal detection is achieved and commercialization.
According to embodiment of the present utility model, the diameter focusing on hot spot is on to the sample less than 1.6 μm.Thus, laser energy can be enable more concentrated, thus greatly can improve the exciting light energy of Raman scattering.In one example, by beam expanding lens carry out more than 7 times expand the focusing realizing above-mentioned hot spot.
Specifically, according to the light path device of above-described embodiment, the laser sent from excitation source 101 by beam expanding lens 102 carry out 7 times expand after, microcobjective 301(20 × apochromatism, flat field, infinite distance metallography microscope object lens are reflexed to through described two to light splitting part 2012 through after narrow band pass filter 2011, and by microcobjective 301 described Laser Focusing formed the hot spot of diameter about 1.6 μm on sample 401 and inspire Raman signal NA0.4).The Raman signal of scattering becomes directional light more again by described Rayleigh optical filter 2013(OD>6 after microcobjective 301) filter Rayleigh scattering, after this converged at the entrance slit place of monochromator 801 (Czerny-Turner) by 45 ° of plane mirrors 501 through pipe mirror 302, and detected by the single photon counter 901 at the exit slit place of monochromator 801.The wavelength of Raman signal light coordinates single photon counter 901 jointly to determine by monochromator 801.
Wherein, microcobjective 301 adopts 20 × apochromatism, flat field, infinite distance metallography microscope object lens, its numerical aperture NA0.4.
As shown in Figure 2, light path device of the present utility model also comprises solid sample imaging system, described solid sample imaging system comprises push-and-pull reflection unit, CCD imaging device, described push-and-pull reflection unit is with carriage, when solid sample need realize focusing, described push-and-pull reflection unit is arranged in and moves to light path, completes defocused, and described push-and-pull reflection unit shifts out light path.
In an embodiment of the present utility model, described push-and-pull reflection unit is push-and-pull plane mirror 601, described CCD imaging device is CCD701, push-and-pull plane mirror 601 for the sample signal from 45 ° of plane mirrors 501 is imaged in CCD701 target surface, to realize the focusing of solid sample.Thus, focusing and the detection of solid sample can be realized.
Wherein, the target surface of CCD701 is equal to the distance of pipe mirror 302 with the entrance slit of monochromator 801 to the distance (herein it is to be appreciated that this place's distance refers to light path) of pipe mirror 302.Easily can realize focusing and the detection of solid sample like this, realize cleverly focusing and the conversion detected by push-and-pull catoptron.
The concrete operations using the light path device of the utility model embodiment to come tracer liquid sample and solid sample are described below respectively.
For fluid sample, as long as open excitation source 101, its minimum light spot focused on after Raman filter set 201 and microcobjective 301 is made roughly to be positioned at the inside of fluid sample).
The focusing of solid sample need by outer light illumination.During solid sample focusing, stationary annular LED illumination lamp on the shell of microcobjective, regulates LED height, makes it maximum in the operating distance place brightness of microcobjective.Push-and-pull plane mirror 601 is pushed in light path, opens CCD701 over cap, regulate the three-dimensional coordinate of sample 401, to make its energy blur-free imaging in CCD701.Particularly, through solid reflection light through microcobjective 301 imaging, outgoing beam is focused on by pipe mirror 302 through Raman filter set 201, to transfer light path through 45 ° of plane mirrors 501, again after push-and-pull plane mirror 601, imaging in CCD701 target surface, making the expression blur-free imaging of solid dielectric in CCD by regulating the height of sample.
Wherein, when regulating sample 401 position, the position that excitation source 101 tentatively guarantees sample 401 can be opened, and then with three-dimensional trim holder, vernier focusing is carried out to sample 401.Before starting detection after focusing, push-and-pull plane mirror 601 is pulled away from light path, with the entrance slit place enabling Raman signal image in monochromator 801.
According to the light path device of the Confocal laser-scanning microscopy instrument of the utility model embodiment, at least tool one of has the following advantages:
All adopt vertical stratification at all turnover positions of light path, therefore light path debug and simply, light path is shorter, thus effectively can reduce the loss of Raman signal, be conducive to expanding sensing range;
In addition, owing to adopting beam expanding lens 102 pairs of laser to expand, micro-effective aperture can be used fully;
Further, owing to using single photon counter 901 as the detector collecting Raman signal, can by suitably increasing the intensity improving tested Raman signal integral time;
Laser Focusing hot spot can reach 1.6um, greatly improves the exciting light energy of Raman scattering;
The microcobjective of bigger numerical aperture increases the collection efficiency of Raman scattering;
This light path is quick in detection Raman signal, efficient, accurately, even the Raman peaks of the water that Raman signal is extremely weak also can detect and obtain;
Above-mentioned solid focusing method can be applied exactly, carry out micro-imaging measurement.
Find through experiment, the resolution of the light path device of the utility model embodiment can reach 0.8um.
According to another aspect of the present utility model, also proposed the Confocal laser-scanning microscopy instrument with above-mentioned arbitrary light path device.
In the description of this instructions, specific features, structure, material or feature that the description of reference term " embodiment ", " some embodiments ", " example ", " concrete example " or " some examples " etc. means to describe in conjunction with this embodiment or example are contained at least one embodiment of the present utility model or example.In this manual, to the schematic representation of above-mentioned term not must for be identical embodiment or example.And the specific features of description, structure, material or feature can combine in one or more embodiment in office or example in an appropriate manner.In addition, when not conflicting, the feature of the different embodiment described in this instructions or example and different embodiment or example can carry out combining and combining by those skilled in the art.
Although illustrate and described embodiment of the present utility model above, be understandable that, above-described embodiment is exemplary, can not be interpreted as restriction of the present utility model, those of ordinary skill in the art can change above-described embodiment, revises, replace and modification in scope of the present utility model.
Claims (13)
1. a light path device for Confocal laser-scanning microscopy instrument, is characterized in that, is included in the following structure that light path sets gradually:
Excitation light source system, described excitation light source system comprises excitation source and beam expanding lens;
Raman filter set, described Raman filter set has side and receives glancing incidence light and the filter sheet structure forming the emergent light vertical with incident light at opposite side;
Microscopic system, described microscopic system comprises the Guan Jing of the microcobjective being arranged at described Raman filter set sample emergent light side respectively and the sample emergent light offside being arranged at described Raman filter set;
Monochromator, from monochromator described in the Raman signal directive that described pipe mirror converges; And
Single photon counter, described single photon counter detects the Raman signal from described monochromator,
Wherein, the laser sent from described excitation source is expanded by described beam expanding lens, through described Raman filter set to described microcobjective, and by described microcobjective, described Laser Focusing is inspired Raman signal on sample, described Raman signal becomes directional light and filters Rayleigh scattering by described Raman filter set again after described microcobjective, after this focus on the entrance slit place of described monochromator through described Guan Jing, and detected by the described single photon counter at exit slit place.
2. the light path device of Confocal laser-scanning microscopy instrument according to claim 1, it is characterized in that, described Raman filter set comprises the vertical narrow band pass filter, two placed to the Rayleigh optical filter of light splitting part and horizontal positioned, described microcobjective is arranged at described two to light splitting part emergent light side, and described Guan Jing is arranged at the emergent light side of described Rayleigh optical filter.
3. the light path device of Confocal laser-scanning microscopy instrument according to claim 2, is characterized in that, described two is 45 ° of plane mirrors to light splitting part.
4. the light path device of Confocal laser-scanning microscopy instrument according to claim 1, it is characterized in that, described light path device also comprises one or many reflection unit, and described reflection unit accepts the Raman signal from described pipe mirror convergence and reflexed to described monochromator.
5. the light path device of Confocal laser-scanning microscopy instrument according to claim 4, is characterized in that, described reflection unit is plane mirror.
6. the light path device of Confocal laser-scanning microscopy instrument according to claim 1, is characterized in that, described microcobjective adopts apochromatism flat field infinite distance metallography microscope object lens.
7. the light path device of Confocal laser-scanning microscopy instrument according to claim 6, is characterized in that, the numerical aperture NA of described microcobjective is more than 0.4.
8. the light path device of Confocal laser-scanning microscopy instrument according to claim 6, it is characterized in that, described light path device also comprises solid sample imaging system, described solid sample imaging system comprises push-and-pull reflection unit, CCD imaging device, described push-and-pull reflection unit is with carriage, and when solid sample need realize focusing, described push-and-pull reflection unit moves in light path, complete defocused, described push-and-pull reflection unit shifts out light path.
9. the light path device of Confocal laser-scanning microscopy instrument according to claim 8, is characterized in that, the CCD target surface in described CCD imaging device to described Guan Jing Distance geometry described in the entrance slit of monochromator equal to the distance of described Guan Jing.
10. the light path device of Confocal laser-scanning microscopy instrument according to claim 1, is characterized in that, the diameter focusing on hot spot is on to the sample less than 1.6 μm.
The light path device of 11. Confocal laser-scanning microscopy instrument according to claim 10, it is characterized in that, described beam expanding lens realizes more than 7 times expanding.
12. Raman spectrometer light path devices according to claim 1, is characterized in that, the entrance slit width of described monochromator system is 32-40 μm.
13. 1 kinds of Confocal laser-scanning microscopy instrument, is characterized in that, comprise the light path device described in any one of right 1-12.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104749162A (en) * | 2015-04-17 | 2015-07-01 | 江苏天瑞仪器股份有限公司 | Confocal Raman spectrometer and light path device thereof |
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| CN104749162A (en) * | 2015-04-17 | 2015-07-01 | 江苏天瑞仪器股份有限公司 | Confocal Raman spectrometer and light path device thereof |
| CN104749162B (en) * | 2015-04-17 | 2018-03-27 | 江苏天瑞仪器股份有限公司 | Confocal laser-scanning microscopy instrument and its light path device |
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