CN210090310U - Optical system of optical tweezers-Raman spectrum single particle detector - Google Patents
Optical system of optical tweezers-Raman spectrum single particle detector Download PDFInfo
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- CN210090310U CN210090310U CN201822256673.XU CN201822256673U CN210090310U CN 210090310 U CN210090310 U CN 210090310U CN 201822256673 U CN201822256673 U CN 201822256673U CN 210090310 U CN210090310 U CN 210090310U
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Abstract
The utility model discloses an optical tweezers-raman spectrum single particle detector optical system relates to optical tweezers technique and raman spectrum field. The utility model discloses a computer, computer electric connection have laser instrument unit, optical coupling and control unit and microscope, fixedly connected with dynamic detection device and spectrum appearance on the microscope, and the laser instrument unit includes infrared laser instrument and visible light laser instrument, and the optical coupling includes the optics flat board with the control unit, and the equal fixed mounting of infrared laser instrument and visible light laser instrument is on the optics flat board, and electric connection computer. The utility model discloses a creatively integrate to an optical system with "optical tweezers" and "micro-Raman spectrum", strengthened current Raman spectrometer system's function, realized the normal position of suspended particle, harmless single particle Raman spectrum and detected, reached "1 +1> 2" effect.
Description
Technical Field
The utility model belongs to the technical field of optical tweezers and micro-Raman spectroscopy, especially, relate to optical tweezers-Raman spectroscopy single particle detector optical system.
Background
A spectral detector (P2) is arranged in an existing commercial micro-Raman spectrometer (sp), and the micro-Raman spectrometer (sp) can only measure fixed tiny samples and cannot measure suspended particles (such as cells, bacteria and aerosol) in liquid or gas state. This is because the raman spectroscopy measurement requires a long time for excitation collection due to the very weak intrinsic characteristic of the raman spectroscopy signal, and single suspended particles in liquid or gas, such as suspended cells (e.g., blood cells) in biological culture fluid, suspended particulate matter PM2.5 in the atmosphere, etc., cannot be measured in situ, because these particles are constantly brownian in their natural state, and the laser cannot be aligned with the excitation for a long time. The current practice is to suck the cells with a microneedle and to fix the aerosol particles with a membrane filter, but both will destroy the original state of the cells or aerosol particles, and the sample requires a complicated preparation process. Particularly, if the cell is sucked by the micro-needle, the fluidity of the protein on the cell membrane is damaged, the cell is damaged, the original state of the cell is changed, and the obtained result is not real. Particularly, cancer cells in medicine have typical "heterogeneity", namely genes of each cell are not completely the same, the research result of a large number of group cells cannot reflect the real situation of each cell, and the single-cell Raman spectrum research is an important means for precise medical treatment and personalized treatment at present. For the above reasons, it is still impossible to accurately study these suspended particles in situ and without damage at the single particle level by raman spectroscopy, which severely limits the development of the fields of life sciences and environments, materials, etc.
SUMMERY OF THE UTILITY MODEL
An object of the utility model is to provide optical tweezers-raman spectrum single particle detector optical system, the utility model discloses a creatively integrate to an optical system with "optical tweezers" and "micro-raman spectrum", strengthened current raman spectrum appearance system's function, realized the normal position of suspended particle, harmless single particle raman spectrum and detected, reached "1 +1> 2" effect.
In order to solve the technical problem, the utility model discloses a realize through following technical scheme:
the utility model relates to an optical tweezers-Raman spectrum single particle detector optical system, which comprises a computer, wherein the computer is electrically connected with a laser unit, an optical coupling and control unit and a microscope, and a dynamic detection device and a spectrometer are fixedly connected on the microscope;
the laser unit comprises an infrared laser and a visible laser;
the optical coupling and control unit comprises an optical flat plate, and the infrared laser and the visible laser are both fixedly arranged on the optical flat plate and are electrically connected with the computer;
the optical flat plate is fixedly provided with a fifth plane reflector matched with the visible light laser, laser is shot into the fifth plane reflector to be reflected and deflected, a first beam expander, a second beam expander, a first electric shutter, a first dichroic mirror, a coupling lens and a first lifting mirror are sequentially arranged on a laser reflection deflection light path, the laser is shot into the first lifting mirror to be reflected and deflected, a second lifting mirror is arranged on the surface of the reflection deflection light path, and the laser is shot into the second lifting mirror to be reflected and deflected to the microscope;
the optical flat plate is fixedly provided with a plate surface on a laser light path of the infrared laser, and a third beam expander, a fourth beam expander, a second electric shutter and a first plane reflector are sequentially arranged on the plate surface;
the microscope is provided with a laser beam inlet, and the light path of the second lifting mirror points to the laser beam inlet.
Furthermore, the microscope comprises a body, wherein a lighting lamp is arranged on the upper portion of the body, a condenser lens, an electric sample stage, an objective lens, a third dichroic mirror, a fourth dichroic mirror and a prism are sequentially arranged in the vertical direction of the lighting lamp, a dynamic detection device and an eyepiece are arranged on the side edge of the body, the central axis of the eyepiece points to the prism, a laser beam inlet is formed in one side edge of the body, a second dichroic mirror is fixedly installed in the laser beam inlet, laser is injected into the second dichroic mirror to be subjected to reflection deflection, a tube lens is arranged on a reflection deflection light path, and the tube lens is located between the third dichroic mirror and the second dichroic mirror;
the laser generates reflection deflection through the second lifting mirror, and a reflection deflection light path penetrates through a laser beam inlet of the microscope and is incident on the second dichroic mirror;
a spectrometer access port and a reflecting mirror are arranged on one side of the fourth dichroic mirror, the reflecting mirror is located in the spectrometer access port, and the spectrometer is installed on the spectrometer access port;
the body is fixedly provided with a high-voltage fluorescence exciter.
Furthermore, the dynamic detection device is a CCD detector and is fixedly installed on a microscope three-ocular interface.
Further, the first electric shutter, the second electric shutter, the sixth beam expanding lens, the spectrometer, the spectrum detector and the electric sample stage are all electrically connected with the computer.
Furthermore, the sixth beam expander is connected to the optical flat plate in a sliding manner through an electric sliding rail.
The utility model discloses following beneficial effect has:
1. the utility model discloses a set up infrared laser instrument, the third beam expander, the fourth beam expander, second electric shutter, first plane mirror, the second plane mirror, fifth beam expander and sixth beam expander constitute the outer light path of "optical tweezers system" jointly, through removing the sixth beam expander along the optical axis, can make optical tweezers control the particle along microscope axial displacement, simultaneously by visible light laser instrument, first beam expander, second beam expander and first electric shutter constitute raman spectrum's excitation light path jointly, infrared bundle closes through first dichroic mirror with raman excitation light beam, through coupling lens, in the leading-in microscope. The optical tweezers and the microscopic Raman spectrum are creatively integrated into an optical system, so that the function of the conventional Raman spectrometer system is enhanced, the in-situ and lossless single-particle Raman spectrum detection of suspended particles is realized, and the effect of 1+1>2 is achieved.
2. The utility model discloses can conveniently realize the dual function of "discernment and sorting", for example, measure the raman spectroscopy of single cell after, according to spectrum display alright discern the kind of cell at once (like cancer cell or the cell of normal tissue), then, directly control contactless, the harmless required cell of branch electing with the optical tweezers to do benefit to further research such as unicellular sequencing.
3. The utility model discloses an optics integrated design has realized the perfect coupling of optical tweezers, micro-raman spectroscopy and microscope imaging, has simplified optical system, has improved the stability and the experimental efficiency of system.
4. The utility model discloses because "optical tweezers" and "raman spectroscopy"'s "excitation light source" adopt same "microscope", "computer" and "microscopic dynamic monitoring", saved the common part in two sets of independent equipment, consequently, greatly reduced the cost.
Of course, it is not necessary for any particular product to achieve all of the above-described advantages at the same time.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to these drawings without creative efforts.
Fig. 1 is a light path structure diagram of the optical system of the optical tweezers-raman spectrum single particle detector of the present invention;
fig. 2 is a schematic structural view of a microscope according to the present invention;
fig. 3 is a schematic diagram of the optical system of the optical tweezers-raman spectroscopy single particle detector of the present invention;
FIG. 4 is a Raman spectrum of a single polystyrene bead trapped in a solution environment with optical tweezers; FIG. 5 shows the Raman spectrum of a single yeast living cell measured systematically.
Detailed Description
The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by a person of ordinary skill in the art without creative efforts belong to the protection scope of the present invention.
In the description of the present invention, it is to be understood that the terms "upper", "lower", "middle", "wall", "inner", "outer", "end", "side", and the like, indicate positional or positional relationships, are merely for convenience in describing the present invention and to simplify the description, and do not indicate or imply that the components or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore should not be construed as limiting the present invention.
Referring to fig. 1, the optical system of the optical tweezers-raman spectroscopy single particle detector comprises a computer, wherein the computer is electrically connected with a laser unit, an optical coupling and manipulation unit, a microscope, a spectrometer and a spectral detector, a dynamic detection device 6 and the spectrometer are fixedly connected to the microscope, the microscope guides laser beams of the laser unit into the microscope through the optical coupling and manipulation unit, and the laser beams are converged on a sample by a microscope objective with a high numerical aperture. The Raman scattering light is received by the same microscope objective, the Raman scattering light is separated from incident light by an optical dichroic mirror filter, the Raman scattering light is guided into a spectrometer through an optical element, is split by the spectrometer and then is projected onto a target surface of a spectrum detector (CCD or PMT) for receiving, and finally, the spectrum data is transmitted to a computer for processing and displaying;
the laser unit comprises an infrared laser H1 and a visible light laser H2;
the optical coupling and control unit comprises an optical flat plate, and the infrared laser H1 and the visible light laser H2 are both fixedly arranged on the optical flat plate and are electrically connected with a computer, so that the control is convenient;
a fifth plane mirror M5 matched with a visible light laser H2 is fixedly installed on the optical flat plate, laser is shot into the fifth plane mirror M5 to be reflected and deflected, a first beam expander L1, a second beam expander L2, a first electric shutter S1, a first dichroic mirror D1, a coupling lens L7 and a first lifting mirror M3 are sequentially arranged on a laser reflection deflection light path, the laser is shot into a first lifting mirror M3 to be reflected and deflected, a second lifting mirror M4 is arranged on the plate surface of the reflection deflection light path, and the laser is shot into a second lifting mirror M4 to be reflected and deflected to point to the microscope;
a third beam expander L3, a fourth beam expander L4, a second electric shutter S2 and a first plane reflector M1 are sequentially arranged on a plate surface on a laser light path fixedly mounted on an infrared laser H1, laser enters a first plane reflector M1 to be reflected and deflected, a fifth beam expander L5, a sixth beam expander L6 and a second plane reflector M2 are fixedly mounted on a plate surface of the reflection and deflection light path, the laser enters a second plane reflector M2 to be reflected and deflected, the reflection and deflection light path points to a first dichroic mirror D1 to be reflected and deflected, two laser beams are combined into a whole in the first dichroic mirror D1 to enter a coupling lens L7, and the first dichroic mirror D1 is bandpass, namely, high reflection of infrared and ultraviolet and high transmission of visible light;
all optical elements are fixed on a 500X 500mm optical flat plate by optical mechanisms, and all the optical elements are on the same horizontal plane and keep collimation and coaxiality.
The beam expansion ratio of the fifth beam expander L5 to the sixth beam expander L6 is 1: 1, adjusting the position of a sixth beam expander L6 to change the longitudinal position of the laser on the sample;
the microscope is provided with a laser beam inlet 14, and the optical path of the second lifting mirror M4 points to the laser beam inlet 14.
Referring to fig. 2, the microscope includes a body 1, a lighting lamp 2 is arranged on the upper portion of the body 1, a condenser lens 3, an electric sample stage 4, an objective lens 5, a third dichroic mirror 10, a fourth dichroic mirror 9 and a prism 8 are sequentially arranged along the vertical direction of the lighting lamp 2, a dynamic detection device 6 and an eyepiece lens 7 are arranged on the side edge of the body 1, the central axis of the eyepiece lens 7 points to the prism 8, a laser beam inlet 14 is arranged on one side edge of the body 1, a second dichroic mirror 12 is fixedly installed in the laser beam inlet 14, laser is incident into the second dichroic mirror 12 to undergo reflection deflection, a tube lens 11 is arranged on a light path of the reflection deflection, and the tube lens 11 is located between the third dichroic mirror 10 and the second dichroic mirror 12;
the laser beam is reflected and deflected through a second lifting mirror M4, and a reflected and deflected light path penetrates through a laser beam inlet 14 of the microscope and is incident on a second dichroic mirror 12;
a spectrometer access port 15 and a reflecting mirror 13 are arranged on one side of the fourth dichroic mirror 9, and the spectrometer is installed in the spectrometer access port 15;
a high-voltage fluorescence exciter 16 is fixedly arranged on the body 1;
the two laser beams are combined through a first dichroic mirror D1, the first dichroic mirror D1 has the functions of high infrared reflection, high ultraviolet and high visible light transmission, the laser beams are guided into a microscope through a coupling lens L7, the laser beams are coupled into a fluorescence channel of the microscope through a second dichroic mirror 12 and are gathered on a sample through an objective lens 5.
Preferably, the dynamic detection device 6 is a CCD detector, and is fixedly mounted on the microscope eyepiece.
Preferably, the first electrically operated shutter S1, the second electrically operated shutter S2, the sixth beam expander L6, the spectrometer sp, the spectrum detector P2 and the electrically operated sample stage 4 are electrically connected to the computer, and the first electrically operated shutter S1 and the second electrically operated shutter S2 are used for controlling the opening and closing of the infrared laser H1 and the visible laser H2.
Preferably, the sixth beam expander L6 is slidably connected to the optical flat via a motorized slide rail.
The infrared laser H1, the third beam expander L3, the fourth beam expander L4, the second electric shutter S2, the first plane mirror M1, the second plane mirror M2, the fifth beam expander L5 and the sixth beam expander L6 jointly form an outer light path of an 'optical tweezers system', and the optical tweezers manipulation particles can be moved along the z axis of the microscope by moving the sixth beam expander L6 along the optical axis.
The visible laser H2, the first beam expander L1, the second beam expander L2, and the first electrically operated shutter S1 together form an excitation optical path of a raman spectrum.
Please refer to fig. 3 to 5 together, fig. 3 is a schematic diagram of the optical path of the optical tweezers-raman spectroscopy single particle detection system of the present invention, fig. 4 is a raman spectrum of a single polystyrene bead trapped by the optical tweezers in the solution environment, fig. 5 is a raman spectrum of a single yeast living cell measured by the system, the measurement effect is good, and by using the present invention, the dual functions of "identification and sorting" can be conveniently realized, for example, after measuring the raman spectrum of a single cell, the cell type can be identified immediately according to the spectrum display, and then, the required cell can be directly sorted out without damage by the optical tweezers operation, so as to facilitate the further research of single cell sequencing.
The preferred embodiments of the present invention disclosed above are intended only to help illustrate the present invention. The preferred embodiments are not intended to be exhaustive or to limit the invention to the precise embodiments disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and its practical applications, to thereby enable others skilled in the art to best understand the invention for and utilize the invention. The present invention is limited only by the claims and their full scope and equivalents.
Claims (5)
1. The optical system of the optical tweezers-Raman spectrum single particle detector comprises a computer and is characterized in that the computer is electrically connected with a laser unit, an optical coupling and control unit and a microscope, and a dynamic detection device (6) and a spectrometer are fixedly connected to the microscope;
the laser unit comprises an infrared laser (H1) and a visible laser (H2);
the optical coupling and manipulation unit comprises an optical flat plate, and the infrared laser (H1) and the visible laser (H2) are both fixedly arranged on the optical flat plate and are electrically connected with the computer;
the optical flat plate is fixedly provided with a fifth plane reflector (M5) matched with a visible light laser (H2), the laser is injected into the fifth plane reflector (M5) to be reflected and deflected, a first beam expander (L1), a second beam expander (L2), a first electric shutter (S1), a first dichroic mirror (D1), a coupling lens (L7) and a first lifting mirror (M3) are sequentially arranged on a laser reflection deflection light path, the laser is injected into a first lifting mirror (M3) to be reflected and deflected, a second lifting mirror (M4) is arranged on the plate surface of the reflection deflection light path, and the laser is injected into a second lifting mirror (M4) to be reflected and deflected to point to the microscope;
a third beam expander (L3), a fourth beam expander (L4), a second electric shutter (S2) and a first plane reflector (M1) are sequentially arranged on a plate surface on a laser light path fixedly mounted with an infrared laser (H1), laser is irradiated into the first plane reflector (M1) to be reflected and deflected, a fifth beam expander (L5), a sixth beam expander (L6) and the second plane reflector (M2) are fixedly mounted on the plate surface of the reflected and deflected light path, the laser is irradiated into the second plane reflector (M2) to be reflected and deflected, and the reflected and deflected light path points to a first dichroic mirror (D1) to be reflected and deflected and is irradiated into a coupling lens (L7);
the microscope is provided with a laser beam inlet (14), and the light path of the second lifting mirror (M4) points to the laser beam inlet (14).
2. The optical system of the optical tweezers-Raman spectrum single particle detector according to claim 1, wherein the microscope comprises a body (1), a lighting lamp (2) is arranged on the upper portion of the body (1), a condenser lens (3), an electric sample stage (4), an objective lens (5), a third dichroic mirror (10), a fourth dichroic mirror (9) and a prism (8) are sequentially arranged in the vertical direction of the lighting lamp (2), a dynamic detection device (6) and an eyepiece (7) are arranged on the side edge of the body (1), the central axis of the eyepiece (7) points to the prism (8), a laser beam inlet (14) is arranged on one side edge of the body (1), a second dichroic mirror (12) is fixedly arranged in the laser beam inlet (14), laser enters the second dichroic mirror (12) to generate reflection deflection, and a tube mirror (11) is arranged on a reflection deflection light path, the tube mirror (11) is arranged between the third dichroic mirror (10) and the second dichroic mirror (12);
the laser generates reflection deflection through a second lifting mirror (M4), and a reflection deflection light path penetrates through a laser beam inlet (14) of the microscope and is incident on a second dichroic mirror (12);
a spectrometer access port (15) and a reflecting mirror (13) are arranged on one side of the fourth dichroic mirror (9), the reflecting mirror (13) is positioned in the spectrometer access port (15), and the spectrometer is installed on the spectrometer access port (15);
the body (1) is fixedly provided with a high-voltage fluorescence exciter (16).
3. The optical system of the optical tweezers-raman spectroscopy single particle detector according to claim 1, wherein the dynamic detection device (6) is a CCD detector and is fixedly mounted on a microscope trio-ocular interface.
4. The optical tweezers-raman spectroscopy single particle detector optical system according to claim 1, wherein the first motorized shutter (S1), the second motorized shutter (S2), the sixth beam expander lens (L6), the spectrometer, the spectral detector, and the motorized sample stage (4) are all in electrical communication with a computer.
5. The optical system of claim 1, wherein the sixth beam expander (L6) is slidably connected to the optical flat via a motorized sled.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201822256673.XU CN210090310U (en) | 2018-12-29 | 2018-12-29 | Optical system of optical tweezers-Raman spectrum single particle detector |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201822256673.XU CN210090310U (en) | 2018-12-29 | 2018-12-29 | Optical system of optical tweezers-Raman spectrum single particle detector |
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| CN210090310U true CN210090310U (en) | 2020-02-18 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113820301A (en) * | 2021-11-25 | 2021-12-21 | 之江实验室 | Method and device for identifying microorganism species by using Raman optical tweezers |
-
2018
- 2018-12-29 CN CN201822256673.XU patent/CN210090310U/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113820301A (en) * | 2021-11-25 | 2021-12-21 | 之江实验室 | Method and device for identifying microorganism species by using Raman optical tweezers |
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Granted publication date: 20200218 Termination date: 20201229 |