WO2018076244A1

WO2018076244A1 - Ellipsoidal mirror-based biofluorescence capturing structure and capturing method

Info

Publication number: WO2018076244A1
Application number: PCT/CN2016/103599
Authority: WO
Inventors: 吴彦林
Original assignee: 西安精英光电技术有限公司
Priority date: 2016-10-27
Filing date: 2016-10-27
Publication date: 2018-05-03

Abstract

An ellipsoidal mirror-based biofluorescence capturing structure and a capturing method. The capturing structure comprises an ellipsoidal mirror (1), a capillary (3), and a fluorescence output optical fiber (6). The capillary (3) is mounted at a proximal focus of the ellipsoidal mirror (1). A reagent-marked biological test sample is mounted in the capillary (3). An extremity of the fluorescent output optical fiber (6) is arranged at a distal focus of the ellipsoidal mirror (1). When in use, a light beam is focused at the center of the capillary (3), the marked matter in the capillary (3) is excited to emit a light, thus producing fluorescence, the light source of the fluorescence radiates fluorescent photons to spaces in all directions, at which time, the radiated photons are reflected, light rays emitted from the near focus are converged to the distal focus, and the fluorescence output optical fiber (6) arranged at the distal focus of the ellipsoidal mirror (1) guides the fluorescence into the fluorescence output optical fiber (6) and then transmits same. The capturing structure is structurally compact, increases fluorescence capturing efficiency to the greatest extent, and reduces error of an optical system.

Description

Bio-fluorescence collecting structure and collecting method based on ellipsoidal mirror

[Technical Field]

The invention belongs to the field of optoelectronic technology, and in particular to the field of laser bio-detection.

【Background technique】

Photochemical bioassay is a fast-growing industry in recent years. Photoluminescence is very broad in the fields of genetic detection sequencing, pathogen recognition, and newborn screening because of its ease of use, low consumables, high detection speed, and easy expansion. In the future, large foreign medical device manufacturers have basically monopolized the production and licensing of the above devices.

Domestic biomedical device technology has also begun to get involved in this field in recent years, but there are still big differences in equipment scale, reliability, and service life. On the basis of imitating imported equipment, engineers and technicians have found that many equipments are early because of stereotypes. At that time, there was almost room for optimization and improvement in terms of technical level or process limitations. This also provided domestic manufacturers with the advantages of late-stage technology. For this market demand, pioneering use of aspheric mirrors in the field of laser bio-detection Perform back-fluorescence collection.

Conventional fluorescence acquisition optics, using planar, spherical, parabolic mirrors, or directly using a spherical lens system as an optical shaping device, has the advantage that the above devices are easy to implement in the process because of conventional optical cold working, planar, spherical, parabolic mirrors. And the ball lens processing technology is summarized by the long-term process, these optical components are mature in processing into debugging technology.

[Summary of the Invention]

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.

The invention adopts the following technical solutions:

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.

The first oblique hole disposed at the bottom of the ellipsoidal mirror is coaxial with the beam direction of the laser fiber collimator.

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.

The capillary is disposed perpendicular to a central section of the ellipsoidal mirror.

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. At this time, 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.

Further, 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.

Compared with the prior art, the present invention has at least the following beneficial effects: 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. In use, 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. At this time, the radiated photons are reflected, and all the light emitted by the near focus is emitted. Converging at the far focus, at the far focus of the ellipsoidal mirror, a fluorescent output fiber is provided to guide the fluorescence into the fiber for backward transmission. The invention has compact structure, smart and reasonable design, maximizes fluorescence collection efficiency and reduces optical system error.

[Description of the Drawings]

The drawings described herein are provided to provide a further understanding of the invention, and are not a limitation of the invention.

Figure 1 is a schematic view showing the structure of the assembly of the present invention.

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.

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.

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.

【detailed description】

The present invention will be described in detail with reference to the accompanying drawings and the accompanying drawings.

Referring to FIG. 1 , 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.

Referring to FIG. 2 and FIG. 4, 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. Specifically, 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 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.

Referring to FIG. 3, 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. Specifically, 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. In the present invention, 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. In the present invention, 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 narrow-band filter 5 is a quartz sheet coated by a multi-layer coating, and its main function is to ensure that the wavelength of the fluorescent light passes through and filters out the wavelength of the excitation light. The multi-coated quartz flakes are optimized by the wavelength of the fluorescence wavelength and are mounted on the long axis of the ellipsoid. The mounting surface is perpendicular to the long axis of the ellipsoid and the front end of the fluorescent output fiber.

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. Specifically, 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. Fluorescent photons, surrounded by a half-packed ellipsoidal mirror 1 and placed at a focus (near focus) of the ellipsoidal mirror 1, at which point the radiated photons are reflected, ellipsoidal reflections Mirror 1 can concentrate all the light emitted by the focus to another focus (far focus) because of its geometric characteristics. Further, on the ellipsoidal mirror 1, there is a first inclined hole 11 for transmitting the super-transmitted excitation laser light to prevent the excitation light from affecting the subsequent optical path, and the other focus (distal focus) of the ellipsoidal mirror 1 is An end face of a fluorescent output fiber 6 having a large core diameter and a large value is used to transmit the fluorescent light to the rear of the optical fiber. In order to further reduce the interference of the excitation light, a narrow band filter which can only transmit the fluorescent band is placed in front of the optical fiber end. 5. The compact structure and ingenious design ensure maximum fluorescence collection efficiency and reduced optical system errors.

Compared with the prior art, the invention has the following advantages:

1. The invention has the advantages of compact structure, reasonable design and simple assembly, and the size of the fluorescence collection system is minimized, and the realization is convenient.

2. 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.

3. 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.

4. 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.

In addition to the advantages of the above-mentioned reflection system, 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. Compared with 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.

Claims

A bio-fluorescence collecting structure based on an ellipsoidal mirror, comprising: an ellipsoidal mirror (1), a capillary (3), a laser fiber collimator (4), and a fluorescent output fiber (6), The capillary (3) is mounted on the near focus of the ellipsoidal mirror (1), which is filled with the biological sample to be detected which is calibrated by the reagent, and generates biofluorescence under the action of the laser light source, and the end of the fluorescent output fiber (6) Located at the far focus of the ellipsoidal mirror (1); the excitation laser focuses the beam through the laser fiber collimator to the center of the capillary (3). The target in the capillary is excited by the light to generate fluorescence, and the fluorescent light source is omnidirectional. The space radiates fluorescent photons, which are reflected by the ellipsoidal mirror and all converge on the far focus of the ellipsoid and are finally output by the fluorescent output fiber.
The ellipsoidal mirror-based bio-fluorescence collecting structure according to claim 1, wherein the ellipsoidal mirror is an aspherical mirror that is optically cold-processed and polished with silver in the inner ellipsoidal surface. .
The ellipsoidal mirror-based bio-fluorescence collecting structure according to claim 1 or 2, wherein the bottom of the ellipsoidal mirror is provided with a first inclined hole (11), and the first inclined hole ( 11) The center line passes through the ellipsoid near the focus and is at an angle to the long axis of the ellipsoid.
The ellipsoidal mirror-based bioluminescence collecting structure according to claim 3, characterized in that: the first inclined hole (11) and the laser fiber collimator (4) disposed at the bottom of the ellipsoidal mirror The beam direction is coaxial.
An ellipsoidal mirror-based bioluminescence collecting structure according to claim 1, characterized in that the bottom of the ellipsoidal mirror (1) is provided with a first introduction hole for introducing a capillary (3) (13) And a first extraction hole (15) leading out of the capillary, the first introduction hole (13) and the first extraction hole (15) passing through the near focus of the ellipsoid and perpendicular to the long axis direction of the ellipsoid.
The ellipsoidal mirror-based bioluminescence collecting structure according to claim 1, characterized in that the capillary (3) is disposed perpendicular to a central section of the ellipsoidal mirror (1).
The ellipsoidal mirror-based bioluminescence collecting structure according to claim 1, wherein a narrow band filter capable of transmitting only a fluorescent band is further disposed in front of the end of the fluorescent output fiber (6). Sheet (5) to reduce the interference of the excitation light.
The ellipsoidal mirror-based bio-fluorescence collecting structure according to claim 7, wherein the narrow-band filter (5) is a quartz sheet which is coated by a plurality of layers and transmits fluorescence through a wavelength. The wavelength is center-optimized and mounted on the long axis of the ellipsoid, and the mounting surface is perpendicular to the long axis of the ellipsoid.
The ellipsoidal mirror-based bio-fluorescence collecting structure according to claim 1, wherein the end face of the fluorescent output fiber (6) is perpendicular to the long axis of the ellipsoidal mirror (1), and the fluorescent wavelength is plated. Antireflection film.
The ellipsoidal mirror-based bioluminescence collecting structure according to claim 1, wherein the output beam focus point of the laser fiber collimator (4) is located at a central section of the ellipsoidal mirror (1). The intersection with the capillary.
The ellipsoidal mirror-based bioluminescence collecting structure according to claim 1, wherein the output beam focus point of the laser fiber collimator (4) is located on the ellipsoidal mirror (1) reflecting surface. Near focus.
The ellipsoidal mirror-based bio-fluorescence collecting structure according to claim 1, characterized in that the beam waist size of the laser fiber collimator (4) is similar to the inner diameter of the capillary (3) to ensure uniformity. The target within the capillary is illuminated.
An ellipsoidal mirror-based bioluminescence collecting structure according to any one of claims 1 to 12, characterized in that the ellipsoidal mirror (1) is mounted in a mirror mount (2), The mirror mount (2) adjusts the mounting angle and position of the ellipsoidal mirror (2) through the top ram and spring member of the device mounting position.
The ellipsoidal mirror-based bio-fluorescence collecting structure according to claim 13, characterized in that the mirror mount (2) is a metal structure made of brass or stainless steel, and the surface is blackened.
An ellipsoidal mirror-based bioluminescence collecting structure according to any one of claims 3 to 12, characterized in that the ellipsoidal mirror (1) is mounted in the mirror mount (2), The mirror mount (2) is provided with a second inclined hole (21) coaxially and in communication with the first inclined hole (11) of the ellipsoidal mirror (1).
An ellipsoidal mirror-based bioluminescence collecting structure according to any one of claims 4 to 12, characterized in that the ellipsoidal mirror (1) is mounted in the mirror mount (2), The mirror mount (2) is provided with a second introduction hole (23) and a second extraction hole (25), the second introduction hole (23) and the second extraction hole (25) and the ellipsoidal mirror An introduction hole (13) and a first extraction hole (15) are coaxially and in communication with each other for introducing and extracting the capillary (3).
An acquisition method of an ellipsoidal mirror-based bioluminescence acquisition structure according to claim 1, characterized in that the excitation laser focuses the beam on the center of the capillary (3) through the laser fiber collimator (4). The target in the capillary (3) is excited by the light to generate fluorescence. Because of the mechanism and characteristics of the fluorescence, the laser irradiation point is the fluorescent light source, which radiates the fluorescent photon to the omnidirectional space, and the ellipsoidal mirror (1) The light source is surrounded and the light source is placed in the near focus of the ellipsoidal mirror (1). At this time, the radiated photons are reflected, and the ellipsoidal mirror (1) concentrates all the light emitted by the near focus on the ellipsoid ( The far focus of 1) is finally output by the fluorescent output fiber (6).
The method for collecting an ellipsoidal mirror-based bio-fluorescence collecting structure according to claim 17, characterized in that the ellipsoidal mirror (1) is provided with a first inclined hole (11), which is super-transmissive. The excitation laser is transmitted out to prevent the excitation light from affecting the subsequent optical path.
The method for collecting a bio-fluorescence collecting structure based on an ellipsoidal mirror according to claim 17, wherein a fluorescent wave-emitting band is further disposed in front of the end of the fluorescent output fiber (6). The narrow band filter (5) reduces the interference of the excitation light as the beam passes.