WO2018076244A1 - Structure de capture de bio-fluorescence fondée sur un miroir ellipsoïdal et procédé de capture - Google Patents

Structure de capture de bio-fluorescence fondée sur un miroir ellipsoïdal et procédé de capture Download PDF

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Publication number
WO2018076244A1
WO2018076244A1 PCT/CN2016/103599 CN2016103599W WO2018076244A1 WO 2018076244 A1 WO2018076244 A1 WO 2018076244A1 CN 2016103599 W CN2016103599 W CN 2016103599W WO 2018076244 A1 WO2018076244 A1 WO 2018076244A1
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WO
WIPO (PCT)
Prior art keywords
ellipsoidal mirror
mirror
capillary
fluorescence
fluorescent
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PCT/CN2016/103599
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English (en)
Chinese (zh)
Inventor
吴彦林
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西安精英光电技术有限公司
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Priority to PCT/CN2016/103599 priority Critical patent/WO2018076244A1/fr
Publication of WO2018076244A1 publication Critical patent/WO2018076244A1/fr

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    • 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
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • 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

Definitions

  • the invention provides a bio-fluorescence collecting structure and an acquisition method based on an ellipsoidal mirror, which has a compact structure, a clever and rational design, and maximizes the fluorescence collection efficiency and reduces the optical system error.
  • a bioluminescent collection structure based on an ellipsoidal mirror comprising an ellipsoidal mirror, a capillary, a laser fiber collimator, and a fluorescent output fiber, the capillary being mounted at a near focus of the ellipsoidal mirror, the capillary being mounted therein
  • the biological sample to be detected calibrated by the reagent generates biofluorescence under the action of a laser light source, and the end of the fluorescent output fiber is located on the ellipsoid
  • the far focus of the mirror; the excitation laser focuses the beam on the center of the capillary through the laser fiber collimator, the target object in the capillary is excited by the light, and generates fluorescence, and the fluorescent light source generates a fluorescent photon to the omnidirectional space, and the fluorescent photon is After being reflected by the ellipsoidal mirror, all of them converge on the far focus of the ellipsoid and are finally output by the fluorescent output fiber.
  • the ellipsoidal mirror is an aspherical mirror that is optically cold worked and polished with silver in the inner ellipsoid.
  • the bottom of the ellipsoidal mirror is provided with a first inclined hole, and the center line of the first inclined hole passes through the ellipsoid near the focus and is at an angle with the long axis of the ellipsoid.
  • a bottom of the ellipsoidal mirror is provided with a first introduction hole for introducing a capillary and a first extraction hole for taking out a capillary, the first introduction hole and the first extraction hole are close to the ellipsoid and are perpendicular to the ellipsoid Long axis direction.
  • a narrow band filter that can only transmit through the fluorescent band is placed in front of the end of the fluorescent output fiber to reduce excitation light interference.
  • the narrow-band filter is a quartz sheet which is coated by a multi-layer coating, and the transmission wavelength is optimized with the fluorescence wavelength as the center, and is mounted on the long axis of the ellipsoid, and the mounting surface is perpendicular to the long axis of the ellipsoid.
  • the end face of the fluorescent output fiber is perpendicular to the long axis of the ellipsoidal mirror and is plated with a fluorescent wavelength antireflection film.
  • the output beam focus point of the laser fiber collimator is located at the intersection of the central section of the ellipsoidal mirror and the capillary.
  • the output beam focus point of the laser fiber collimator is located at a near focus of the reflecting surface of the ellipsoidal mirror.
  • the beam waist size of the laser fiber collimator is similar to the inner diameter of the capillary to ensure uniform illumination of the target within the capillary.
  • the ellipsoidal mirror is mounted in a mirror mount that adjusts the mounting angle and position of the ellipsoidal mirror through a top ram and a spring member at the device mounting location.
  • the mirror mount is made of metal and is made of brass or stainless steel with a blackened surface.
  • the ellipsoidal mirror is mounted in the mirror mount, and the mirror mount is provided with a second oblique hole coaxially and in communication with the first oblique hole of the ellipsoidal mirror.
  • the ellipsoidal mirror is mounted in a mirror mount, the mirror mount is provided with a second introduction hole and a second extraction hole, and the second introduction hole and the second extraction hole and the first of the ellipsoidal mirror
  • the introduction hole and the first extraction hole are coaxially and in communication with each other for introducing and extracting the capillary.
  • a bio-fluorescence acquisition method based on an ellipsoidal mirror the excitation laser focuses the beam on the center of the capillary through a laser fiber collimator, and the target object in the capillary is excited by the light to generate fluorescence, because of the mechanism and characteristics of the fluorescence, the laser
  • the illuminating point is a fluorescent illuminating light source that radiates fluorescent photons into the omnidirectional space.
  • the ellipsoidal mirror surrounds the light source and places the light source in the near focus of the ellipsoidal mirror.
  • the radiated photons are reflected, and the ellipses are The spherical mirror concentrates all the light emitted by the near focus onto the far focus of the ellipsoid and is finally output by the fluorescent output fiber.
  • the ellipsoidal mirror is provided with a first inclined hole to transmit the excess transmitted excitation laser light to prevent the excitation light from affecting the subsequent optical path.
  • a narrow-band filter that can only transmit through the fluorescent band is placed in front of the end of the fluorescent output fiber, and the excitation light is reduced when the light beam passes.
  • the collecting structure of the present invention comprises an ellipsoidal mirror, a capillary tube, and a fluorescent output optical fiber, the capillary being mounted at a near focus of the ellipsoidal mirror, the capillary being mounted therein
  • the reagent-calibrated biological sample to be detected the end of the fluorescent output fiber being located at a far focus of the ellipsoidal mirror.
  • the beam is focused on the center of the capillary, and the target in the capillary is excited to produce fluorescence.
  • the fluorescent source radiates the fluorescent photon into the omnidirectional space.
  • the invention has compact structure, smart and reasonable design, maximizes fluorescence collection efficiency and reduces optical system error.
  • Figure 1 is a schematic view showing the structure of the assembly of the present invention.
  • FIG. 2 is a schematic structural view of an ellipsoidal mirror, wherein (a) is a plan view, (b) is a perspective view, (c) is a cross-sectional view, and (d) is a cross-sectional view taken at another angle.
  • FIG. 3 is a schematic view of a mirror mount, wherein (a) is a plan view, (b) is a perspective view, (c) is a cross-sectional view, and (d) is a cross-sectional view taken at another angle.
  • FIG. 4 is a schematic view showing a combination of an ellipsoidal mirror and a mount, wherein (a) is a plan view, (b) is a perspective view, (c) is a cross-sectional view, and (d) is a cross-sectional view taken at another angle.
  • 1-ellipsoidal mirror 2-mirror mount; 3-capillary; 4-laser fiber collimator; 5-narrowband filter; 6-fluorescent output fiber.
  • the present invention provides a bio-fluorescence collecting structure based on an ellipsoidal mirror, including an ellipsoidal mirror 1 , a mirror mount 2 , a capillary 3 , a laser fiber collimator 4 , and narrow-band filtering. Sheet 5, fluorescent output fiber 6.
  • the ellipsoidal mirror 1 is an aspherical mirror which is optically cold processed and polished with silver on the inner ellipsoid surface, and the scattering fluorescence is collected by the reflection characteristic of the ellipsoid surface.
  • the ellipsoidal mirror 1 is mounted in the mirror mount 2 and formed integrally by an optical fixing glue.
  • the substrate material of the ellipsoidal mirror 1 is based on an optical glass material, and the appearance is a cylindrical structure with an ellipsoidal groove, which is based on optical cold working or lamination, and is internally optically polished.
  • a silver-plated reflective layer a silver-plated reflective layer; a first oblique hole 11 is formed beside the center of the bottom of the ellipsoidal surface, and the center line of the first inclined hole 11 passes through the ellipsoidal surface and has a close angle with the long axis; the near focus is perpendicular to the long axis In the direction
  • a first introduction hole 13 and a first extraction hole 15 for introduction and extraction of the capillary are respectively opened.
  • the ellipsoidal mirror 1 utilizes the characteristics of two focal points of the ellipsoidal surface (the light emitted by one focus illuminates the ellipsoid, and the reflected light must converge on the other focus of the ellipsoid).
  • the fluorescence detection area of the capillary is taken as a The light source is placed at one of the focal points of the ellipsoidal mirror, and the end face of the fluorescent output fiber is placed at another focus of the ellipsoidal mirror.
  • the mirror mount 2 is a metal structure of a glass optical component (ie, the aforementioned ellipsoidal mirror 1), which is made of brass or stainless steel and has a black surface.
  • the main function is The angle and position of the clamping of the glass optics are ensured, and the mounting angle and position of the optical structure are adjusted by the top ram and the spring member, and the mirror mount 2 also functions to protect the optical structure and the reticle.
  • the inner groove size of the mirror mount 2 closely matches the cylindrical size of the ellipsoidal mirror 1; the bottom has a second inclined hole 21, and the first inclined hole 11 at the bottom of the ellipsoidal mirror 1 Coaxial and connected; a second introduction hole 23 and a second extraction hole 25 for introducing and withdrawing a capillary tube in a direction close to the major axis of the ellipsoidal lens and perpendicular to the long axis, the second introduction hole 23 and the first introduction hole
  • the holes 13 are coaxial and communicated, and the second extraction holes 25 are coaxial with and communicate with the second introduction holes 15.
  • the capillary 3 is used for detecting a biological fluid having an inner diameter of only several tens of micrometers.
  • the capillary 3 is introduced as a standard device, and a fluorescent excitation point on the capillary is used as a light source of the present invention.
  • the capillary is mounted at a near focus of the ellipsoidal mirror perpendicular to the central section of the ellipsoidal mirror.
  • the laser fiber collimator 4 is an optical device for collimating a single mode fiber output.
  • the laser fiber collimator 4 is introduced as a standard device, mainly as an excitation source for laser biodetection.
  • the output beam focus point of the laser fiber collimator 4 is located just at the center of the ellipsoidal mirror 1 and the focus of the capillary 3, which is also the near focus of the ellipsoidal reflecting surface, the beam direction and the first of the bottom of the ellipsoidal mirror
  • the oblique hole is coaxial.
  • the fluorescent output fiber 6 is a quartz or resin fiber with a large core diameter and a large numerical aperture, the end surface is at a position of an ellipsoidal far focus, and the end surface is perpendicular to the long axis of the ellipsoidal mirror, and the fluorescent wavelength is coated with an antireflection film. The effect is to couple the fluorescence output from the mirror and conduct it to a fluorescence detection device or other optical shaping structure.
  • the present invention collects laser-excited fluorescence into an optical fiber through an ellipsoidal mirror.
  • the excitation laser is output by the semiconductor pigtail laser, and the excitation light beam is focused by the laser fiber collimator 4 at the center of the capillary 3.
  • the beam waist size of the laser fiber collimator 4 is similar to the capillary inner diameter to ensure uniform illumination.
  • the biological sample to be detected is marked by the reagent in the capillary, and the target in the capillary is excited by the light to generate fluorescence. Because of the mechanism and characteristics of the fluorescence, it can be considered that the irradiation point of the laser is a fluorescent light source, and the light source radiates to the omnidirectional space.
  • the compact structure and ingenious design ensure maximum fluorescence collection efficiency and reduced optical system errors.
  • the present invention is a reflective optical path collecting system.
  • the reflective optical path has several obvious advantages: the reflective system has a relatively high space utilization rate and is convenient for miniaturization; the reflective focusing system can avoid chromatic aberration that cannot be eliminated by the refractive system.
  • the optical path system system is very convenient to transplant, the fluorescence of different wavelengths can be completely transmitted without color difference; the excitation light introduced by the reflective optical system is weak, and the excitation light is a very good collimation single-mode light output, which can be used for transmission. Directly filter out most of the way The excitation light source interferes.
  • the excitation light input of the present invention is monochromatic light introduced by a single-mode optical fiber, which reduces the difficulty of debugging the moving optical path compared to the spatial optical path input.
  • the single-mode optical fiber beam has good quality and less stray light, which is beneficial to the detection system to improve the letter. Noise ratio.
  • the invention uses the optical fiber output fluorescence, can be used as a standard detection module, and is connected to the subsequent optical path through a standard optical fiber interface, which greatly reduces the difficulty of user debugging.
  • the ellipsoidal mirror of the present invention has the following special features: 1.
  • the ellipsoidal mirror is irradiated onto the ellipsoid by a light emitted from a focus, and the reflected light must be concentrated on the ellipsoid.
  • Another focus this is the basic physical law of ellipsoidal reflection, which is very suitable for collecting the weak light from a very small scattering source to a focus, increasing the focus energy density, enhancing the signal strength, and improving the signal-to-noise ratio of the detection system.
  • planar, spherical, and parabolic reflection systems it can be directly coupled into optical fibers such as small aperture optics.
  • Planar, spherical, and parabolic reflection systems require lens systems for shaping and focusing; 2.
  • the theoretical collection efficiency of conventional lens systems can only be achieved. 50%, actually only 10% or even lower, to achieve high collection efficiency can only increase the lens numerical aperture, which requires the lens close enough to the light source and a very short focal length, but close to the light source will bring assembly difficulties, shielding incident Light path, short focal length means that a large spherical aberration will be introduced, even if these two problems are overcome, Efficiency is still very low, and the ellipsoidal mirror may be a fluorescent light source surrounded by half or even all-around, reflex system does not introduce spherical aberration and chromatic aberration.

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  • Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

L'invention concerne une structure de capture de bio-fluorescence fondée sur un miroir ellipsoïdal et un procédé de capture. La structure de capture comprend un miroir ellipsoïdal (1), un capillaire (3) et une fibre optique de sortie de fluorescence (6). Le capillaire (3) est fixé au niveau d'un foyer proximal du miroir ellipsoïdal (1). Un échantillon d'essai biologique marqué par un réactif est fixé dans le capillaire (3). Une extrémité de la fibre optique de sortie de fluorescence (6) est agencée au niveau d'un foyer distal du miroir ellipsoïdal (1). Lors de l'utilisation, un faisceau lumineux est focalisé au centre du capillaire (3), la matière marquée dans le capillaire (3) est excitée afin d'émettre une lumière, produisant ainsi une fluorescence, la source de lumière de la fluorescence rayonne des photons fluorescents vers des espaces dans toutes les directions, et, à ce moment, les photons rayonnés sont réfléchis, les rayons lumineux émis par le foyer proche sont convergents vers le foyer distal et la fibre optique de sortie de fluorescence (6) agencée au niveau du foyer distal du miroir ellipsoïdal (1) guide la fluorescence dans la fibre optique de sortie de fluorescence (6) puis transmet cette dernière. La structure de capture est structurellement compacte, permet d'augmenter l'efficacité de capture de fluorescence au maximum et de réduire l'erreur d'un système optique.
PCT/CN2016/103599 2016-10-27 2016-10-27 Structure de capture de bio-fluorescence fondée sur un miroir ellipsoïdal et procédé de capture WO2018076244A1 (fr)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108760690A (zh) * 2018-05-23 2018-11-06 哈尔滨工业大学 椭球反射镜聚焦环形孔径照明光学谐波生成激发方法
CN113624733A (zh) * 2021-06-25 2021-11-09 港湾之星健康生物(深圳)有限公司 高效散射光聚光组件

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4871249A (en) * 1987-07-10 1989-10-03 Medical Research Council Light collecting device with chamber including ellipsoidal surface and spherical surface
WO2003085387A1 (fr) * 2002-04-04 2003-10-16 Biomed Photonics Co., Ltd. Dispositif de detection d'une epifluorescence induite par laser
CN2786619Y (zh) * 2005-05-20 2006-06-07 中国科学院安徽光学精密机械研究所 单粒子气溶胶紫外荧光分析光学测量装置
CN102253021A (zh) * 2011-05-03 2011-11-23 杭州电子科技大学 一种线形激光光束增强型重金属含量检测方法
CN103941381A (zh) * 2014-04-04 2014-07-23 浙江卷积科技有限公司 一种三维空间内微弱光收集器
CN104931471A (zh) * 2015-06-08 2015-09-23 北京大学 大气活性自由基的激光诱导荧光检测系统

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4871249A (en) * 1987-07-10 1989-10-03 Medical Research Council Light collecting device with chamber including ellipsoidal surface and spherical surface
WO2003085387A1 (fr) * 2002-04-04 2003-10-16 Biomed Photonics Co., Ltd. Dispositif de detection d'une epifluorescence induite par laser
CN2786619Y (zh) * 2005-05-20 2006-06-07 中国科学院安徽光学精密机械研究所 单粒子气溶胶紫外荧光分析光学测量装置
CN102253021A (zh) * 2011-05-03 2011-11-23 杭州电子科技大学 一种线形激光光束增强型重金属含量检测方法
CN103941381A (zh) * 2014-04-04 2014-07-23 浙江卷积科技有限公司 一种三维空间内微弱光收集器
CN104931471A (zh) * 2015-06-08 2015-09-23 北京大学 大气活性自由基的激光诱导荧光检测系统

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108760690A (zh) * 2018-05-23 2018-11-06 哈尔滨工业大学 椭球反射镜聚焦环形孔径照明光学谐波生成激发方法
CN113624733A (zh) * 2021-06-25 2021-11-09 港湾之星健康生物(深圳)有限公司 高效散射光聚光组件

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