The present disclosure is based on the application with the CN filing No. 202010879955.4 filed on Aug. 27, 2020, and claims the priority thereto, and the disclosure of this CN application is incorporated herein in entirety.
The present disclosure relates to the technical field of optical spectra, and specifically relates to an objective lens, an optical imaging device, an optical system, and a detection method of an optical system.
Optical spectrum technologies, such as vibrational spectrum and fluorescence/photoluminescence spectrum, use an optical system to focus excitation beams in a sample region, and parse the physicochemical properties of samples by analyzing variation in frequency and intensity of scattered light (reflected or transmitted light) relative to excitation light. The optical spectrum detection technologies have advantages such as no damage to the samples, low environmental requirements, high sensitivity, and easiness in being carried in existing analysis devices to realize correlation functions, and have wide applications in production and life.
Linearly polarized vertical excitation of a sample is the most common way in optical spectrum technologies. For example, in the fields such as food hygiene detection and public safety detection, the sample is located on a flat substrate surface after pretreatment. A large number of sample molecules that are arranged in disorder on microscopic present uniform distribution on macroscopic, and such averaging effect reduces the sensitivity of the samples to an incident angle and a polarization state of excitation light. However, when the tested samples present periodic arrangement or an amount of the samples is decreased, the polarization state and the excitation angle of the excitation light gradually have significant influences on the spectrum, and bring adverse effects to the detection.
In the related art, the polarization state and the polarization angle of the optical detection device are determined, wherein when the polarization state and the polarization angle of the optical detection device are exactly consistent with the polarization state and the polarization angle required by the samples, the detection of the samples can be realized. However, the samples have different characteristics, and the required polarization states and polarization angles of excitation light are also different. All the optical detection devices in the related art only can detect specific samples.
The inventors found that there is an urgent need in the industry to research and develop an optical spectrum device that can be applied to detection of various samples with different characteristics.
The present disclosure provides an objective lens, an optical imaging device, an optical system, and a detection method of an optical system, for optimizing the structure of the objective lens.
Some embodiments of the present disclosure provide an objective lens, including:
In some embodiments, the optical focusing assembly includes:
In some embodiments, the respective lenses of the lens group are arranged in a row and concave surfaces of the respective lenses face a side where the focal point is located; and, along a direction toward the focal point, refractive indexes of the respective lenses are gradually increased.
In some embodiments, the lens group has at least one of the lenses, and at least one of the concave surface and a convex surface of the lens is a quadratic curved surface.
In some embodiments, the optical focusing assembly includes:
In some embodiments, a curvilinear function of the cambered surface satisfies a parabolic equation.
In some embodiments, the objective lens further includes:
In some embodiments, the total reflection lens includes a hemispheric prism, a convex surface of the hemispherical prism is away from the side where the focal point is located, and a plane of the hemispherical prism faces the side where the focal point is located, and a plane of the total reflection lens is provided with an enhancement layer configured to excite a surface electromagnetic wave mode.
In some embodiments, the enhancement layer includes a first metal layer deposited on the plane of the total reflection lens or a two-dimensional material film layer prepared on the plane of the total reflection lens.
In some embodiments, a material of the first metal layer is selected from one or more of the group consisting of: gold, silver, copper, platinum, and palladium; and alternatively, a material of the two-dimensional material film layer is selected from one or more of the group consisting of: graphene, boron nitride, a transition metal chalcogenide, and a heterojunction composed of a transition metal chalcogenide.
In some embodiments, the total reflection lens includes a hemispherical prism, wherein a convex surface of the hemispherical prism is away from the side where the focal point is located, a plane of the hemispherical prism faces the side where the focal point is located, a semi-reflective and semi-transmissive film layer is deposited on a plane of the total reflection lens, and the semi-reflective and semi-transmissive film layer is configured to make a part of light reflected and make the remaining light transmitted, so as to form a surface total reflection on a surface of the semi-reflective and semi-transmissive film layer away from the housing.
In some embodiments, a material of the semi-reflective and semi-transmissive film layer is selected from one or more of the group consisting of: gold, silver, copper, aluminum, iron, platinum, palladium, and rhodium.
In some embodiments, the semi-reflective and semi-transmissive film layer has a thickness of 5 nanometers˜50 nanometers.
In some embodiments, the total reflection lens uses a hemispherical prism, wherein a convex surface of the hemispherical prism is away from the side where the focal point is located, a plane of the hemispherical prism faces the side where the focal point is located, and a waveguide material layer is deposited on the plane of the total reflection lens.
In some embodiments, the waveguide material layer includes:
In some embodiments, the objective lens further includes:
In some embodiments, the total reflection lens includes:
In some embodiments, an angle of an excitation angle of the optical focusing assembly is 0°˜85°.
In some embodiments, a coverage angle adjustment of the optical focusing assembly is 0.5°˜5°.
In some embodiments, a clear aperture of the objective lens is 1.2 times-10 times a clear aperture of a microscopic objective lens.
Some other embodiments of the present disclosure provide an optical imaging device, including the objective lens provided in any one of the technical solutions of the present disclosure.
In some embodiments, the optical imaging device further includes:
The substrate is made of metal; and alternatively, a third metal layer or a two-dimensional material film layer is provided at a side of the substrate facing the housing.
A material of the third metal layer is selected from one or more of the group consisting of: gold, silver, copper, platinum, and palladium.
A material of the two-dimensional material film layer is selected from one or more of the group consisting of: graphene, boron nitride, a transition metal chalcogenide, and a heterojunction composed of a transition metal chalcogenide.
In some embodiments, the focal point of the optical focusing assembly is located on a surface of the substrate.
In some embodiments, the substrate is located outside the focal point of the optical focusing assembly.
In some embodiments, the substrate is located outside the focal point of the optical focusing assembly, and micro-nanospheres are sandwiched between the substrate and the total reflection lens of the optical focusing assembly.
In some embodiments, the optical imaging device further comprises: nanoparticles, deposited on the substrate, wherein the nanoparticles are located in a focal point region of the optical focusing assembly.
In some embodiments, the optical imaging device further includes:
In some embodiments, the optical imaging device further includes:
In some embodiments, the optical imaging device further includes:
In some embodiments, the polarizer further includes:
In some embodiments, the optical imaging device further includes:
In some embodiments, a polarization direction of light output from the half-wave plate coincides with a fast axis of the spiral wave plate, and the polarizer outputs radially polarized light.
In some embodiments, the polarization direction of the light output from the half-wave plate is perpendicular to the fast axis of the spiral wave plate, and the polarizer outputs angularly polarized light.
In some embodiments, the optical system includes one of the following: an optical sectioning microscope and a two-photon microscope.
Some further embodiments of the present disclosure provide an optical system, including the optical imaging device provided in any one of the technical solutions of the present disclosure.
In some embodiments, the optical system further includes:
In some embodiments, the illumination device includes:
In some embodiments, the illumination device further includes:
In some embodiments, the optical system further includes:
In some embodiments, the optical system further includes an illumination device, wherein the illumination device is configured to provide visible illumination light, and wherein the illumination device includes:
In some embodiments, two sets of optical imaging devices are included, wherein focal points of the two sets of optical imaging devices coincide; and at least one set of the optical imaging devices is mounted to a delay line, so that a light path difference of laser emitted by the two sets of optical imaging devices is adjustable.
In some embodiments, the optical system further includes:
Some further embodiments of the present disclosure provide a detection method of an optical system, implemented by using the optical system provided in any one of the technical solutions in the present disclosure, wherein the detection method of an optical system includes steps of:
In some embodiments, making characteristics of the excitation light satisfy excitation requirements of the sample includes a step of:
In some embodiments, in the annular vector polarized excitation mode, a polarization state of the excitation light is adjusted to one of the following: radial polarization, angular polarization, and linear polarization.
In some embodiments, in the annular vector polarized excitation total reflection mode, the polarization state of the excitation light is adjusted to one of the following: radial polarization and angular polarization.
In some embodiments, in the annular vector polarized excitation KR-SPP mode, the polarization state of the excitation light is adjusted to radial polarization.
In some embodiments, in the annular vector polarized excitation Otto-SPP mode, the polarization state of the excitation light is adjusted to radial polarization.
In some embodiments, in the annular vector polarized excitation planar waveguide mode, the polarization state of the excitation light is adjusted to one of the following: radial polarization, angular polarization, and linear polarization.
In some embodiments, in the annular vector polarized excitation optical sectioning microscope mode, the polarization state of the excitation light is adjusted to one of the following: radial polarization and angular polarization.
In some embodiments, in the annular vector polarized excitation sectioning two-photon microscope mode, the polarization state of the excitation light is adjusted to one of the following: radial polarization and angular polarization
The objective lens provided in the above technical solutions has the housing and the optical focusing assembly, wherein the optical focusing assembly realizes light focusing on the basis of one of the reflection principle and the refraction principle, wherein when the excitation angle of the sample is within the range of 0°˜85°, the optical focusing assembly can realize focusing, therefore, it can be suitable for different optical detection requirements, with a quite wide application range.
The technical solutions provided in the present disclosure are illustrated in more detail below with reference to
Nouns and terms used herein are interpreted.
Optical spectrum technologies, such as vibrational spectrum and fluorescence/photoluminescence spectrum, refers to using the optical systems to focus excitation beams in a sample region and parsing the physicochemical properties of samples by analyzing variation in frequency, intensity and the like of scattered light (reflected or transmitted light) relative to excitation light. The optical spectrum detection technologies have the advantages such as no damage to the samples, low environmental requirements, high sensitivity, and easiness in being carried in existing analysis devices to realize correlation functions, and have wide applications in production and life.
Polarization states include linear polarization, elliptical polarization, circular polarization, and vector polarization. In the above, the vector polarization includes radial polarization, angular polarization, and the like. In the field of optical spectrum detection, the polarization states mainly used are linear polarization, radial polarization, and angular polarization. The inventors found that the polarization state and the excitation angle directly affect the optical spectrum characteristics. For example, in Surface-Enhanced Raman Scattering (SERS) spectroscopy, single-molecule level detection can be realized by optimizing the polarization state and the excitation angle.
Referring to
Dimensions of inner diameters of the first opening 11a and the second opening 11b of the inner cavity 11 can be set as desired by adjusting excitation angle θ and an angle coverage area Δθ. The so-called excitation angle θ refers to an included angle, after a light beam passing through the optical focusing assembly 2, between a central line of the light beam and an optical axis of the objective lens 100, as shown in
The optical focusing assembly 2 can focus light to a set position outside the housing 1 based on the reflection principle (the objective lens 100 is structured as a parabolic reflection-type objective lens 100) and the refraction principle (the objective lens 100 is structured as a refraction-type objective lens 100). It will be described respectively later.
In some embodiments, the excitation angle of the optical focusing assembly 2 has an angle of 0°˜85° The optical focusing assembly 2 has a particularly large range of excitation angle, and can be adapted to excitation light with various excitation angles, thus greatly improving an application range of the objective lens 100.
In some embodiments, the optical focusing assembly 2 has a coverage angle adjustment area Δθ of 0.5°˜5°, as shown in
There are multiple implementation modes for the optical focusing assembly 2, which will be classified and described in detail below.
Referring to
In this case, it can be realized through the lens group 21 that the excitation angle θ of incident light from a last lens (e.g. the fifth lens 215 in lens group of five 21, and a fourth lens 214 in lens group of four 21 described later) can be up to 80°. A wall thickness d of annular light beam can be controlled, by a beam expander 300 and a beam changer 400 which are described later, to be 0.4 mm at the minimum, and in this case, the excitation angle coverage area Δθ of the most marginal light focused on the sample is not more than 2°, and the adjustment precision is very high.
Referring to
Referring to
In the above formula (1), c is a reciprocal of curvature radius of the lens surface, h is a distance from a point on the lens surface to a vertex of the lens surface (namely, vertex of the curved surface where the lens surface is located), and k is a constant of the quadric curved surface.
Referring to
In the situation as shown in
Formulas of quadratic curved surface of the lens group 21 composed of the four lenses are as follows:
In the above formulas (2)˜(4), c is a reciprocal of curvature radius of the lens surface; k is a constant of the quadric curved surface; h is a distance from a point on the curved surface to a vertex of the lens surface; and x and y are coordinates of a point on the curved surface relative to the vertex of the lens surface.
Referring to
Referring to
The objective lens 100 provided in various embodiments of the present disclosure can realize an annular vector polarized excitation total reflection mode, and is particularly suitable for optical detection in a molecular level, in which an object to be detected has a too small dimension and is difficult to observe by naked eyes. The sample 900 to be detected is directly deposited or adsorbed on a plane of the total reflection lens 3. When different samples need to be detected, the total reflection lens 3 is directly replaced. An optical detection method will be described in detail below in conjunction with a light path where the objective lens 100 is located.
Referring to
The objective lens 100 provided in various embodiments of the present disclosure can realize the annular vector polarized excitation total reflection mode, and is particularly suitable for optical detection in a molecular level, in which an object to be detected has a very small dimension and is difficult to observe by naked eyes. The sample 900 to be detected is directly deposited or adsorbed on a plane of the second lens portion 32, wherein when different samples need to be detected, the second lens portion 32 is directly replaced, which can simplify the detection operation. The optical detection method will be described in detail below in conjunction with the light path in which the objective lens 100 is located.
Referring to
In spectrum detection, by placing a sample at the focal point of the objective lens 100, the sample can be effectively excited, and a spectral signal that needs to be detected is produced. In the total internal reflection operation mode, the sample is directly adhered to a sphere center of the hemispherical prism, and the center of the hemispherical prism coincides with the focal point of the objective lens 100. Besides, in the total internal reflection mode of the objective lens 100, the sample also may be disposed at a certain distance from the sphere center of the total internal reflection lens 3, then relatively high excitation efficiency also can be obtained. The distance from the sample to the surface of total reflection lens 3 is generally not more than 20 micrometers.
In the technical solutions provided in various embodiments of the present disclosure, the total reflection lens 3 is used to improve excitation efficiency of excitation light, and energies of excitation beams are concentrated in an annular light beam ring with an extremely narrow wall thickness. By controlling the light beam polarization, the excitation angle and the excitation angle coverage area can be flexibly controlled according to characteristics of the sample. The present disclosure is particularly suitable for applications having particular requirements on the excitation angle and polarization, such as shell-isolated nanoparticle-enhanced spectrum, single-particle-enhanced spectrum, and probe-enhanced spectrum and other fields, and effectively improves the sensitivity of spectrum detection. In addition, the above technical solution reduces the angle control capability achieved by a conventional large and complex corner device in a highly integrated optical device, greatly simplifies the device structure, and can be directly applied to an existing microscopic spectral system.
Referring to
Referring to
The enhancement layer 4 is, for example, a first metal layer deposited on the plane of the total reflection lens 3 or a two-dimensional material film layer prepared on the plane of the total reflection lens 3.
In some embodiments, the material of the first metal layer is selected from one or more of the group consisting of: gold, silver, copper, platinum, and palladium. By providing the first metal layer, a support metal layer may be provided for propagation of surface electromagnetic field.
In some other embodiments, the material of the two-dimensional material film layer is selected from one or more of the group consisting of: graphene, boron nitride, a transition metal chalcogenide, and a heterojunction composed of a transition metal chalcogenide. The heterojunction thickness is 0.2 nanometers˜50 nanometers. By providing the two-dimensional material film, a support material layer can be provided for propagation of surface electromagnetic field.
Referring to
Referring to
Referring to
Referring to
Referring to
In the waveguide mode, a material of the second metal layer 61 is metal such as gold, silver, copper, aluminum, iron, platinum, palladium, and rhodium, with a thickness of 5 nanometers˜50 nanometers. A material of the waveguide medium layer 62 is a medium material transparent under visible and infrared wavelengths, such as silicon dioxide and a doping material thereof, aluminum oxide and a doping material thereof, zinc selenide and a doping material thereof, and cadmium sulfide and a doping material thereof. The waveguide medium layer 62 has a thickness of 50 nanometers˜30 nanometers.
Referring to
In some embodiments, the optical focusing assembly 2 includes a metal plate, the metal plate includes a cambered surface, and the metal plate is integrated with the housing 1.
The metal plate is a plate with sufficiently low surface roughness, specifically a metal mirror surface. Light is directly reflected by the metal plate and then focused at the focal point.
Referring to
The parabolic equation is:
x
2=2=p×y Formula (5)
In the above, x-axis is an optical axis direction of the objective lens 100, y-axis is perpendicular to x-axis, and p represents a distance from the focal point to a directrix, 2 mm≤p≤100 mm. A position of the focal point of the parabolic surface is (0, p/2), and a position of a focal plane 160 is y=2/p. The parabolic surface is a part of y>p/2, wherein a light beam incident along an axial direction is reflected by the parabolic surface and then focused at the focal point, wherein an incident angle ranges from 30° to 90°.
In
Relationships between p, r, d, θ, and Δθ satisfy the following formula (6) and formula (7):
In the above formulas (6) and (7), r is a radius of the annular light beam, and d is a wall thickness of the annular light beam.
The formulas (6) and (7) satisfied by the relationships between p, r, d, θ, and Δθ are applicable to various embodiments in which the optical focusing assembly 2 realizes gathering according to the reflection principle and realizes focusing according to the refraction principle.
The total reflection lens 3 and various film layers provided on the total reflection lens 3 described above are also applicable to the reflection-type objective lens 100. The description is made by way of example below, wherein for the part not explained, reference is made to the description about the part of the refraction-type objective lens 100 in the above.
Referring to
The total reflection lens 3 may be a hemispheric prism, and the hemispheric prism is in an integral structure or a pieced structure. By providing the total reflection lens 3, light forms total reflection on a side of the plane of the total reflection lens 3, which is particularly suitable for scenes requiring optical detection in the total reflection mode.
Referring to
Referring to
The optical imaging device 750 may be any optical imaging device that requires the objective lens 100, such as a spectral analyzer and a microscope.
Referring to
The substrate 8 may be provided regardless of whether the objective lens 100 is provided with the total reflection lens 3 or not. As shown in
Referring to
A material of the nanoparticles 9 located in the focal point region is precious metal such as gold, silver, and copper, and the nanoparticles 9 have a dimension of 20 nanometers˜500 nanometers.
Referring to
Referring to
Specifically, the objective lens 100 of the optical imaging device 750 is provided with the total reflection lens 3, the substrate 8 is spaced apart from the total reflection lens 3, and the substrate 8 is located outside the focal point of the objective lens 100. The sample 900 to be detected is disposed on the surface of the substrate 8 facing the total reflection lens 3.
Referring to
In some other embodiments, the objective lens 100 of the optical imaging device 750 is provided with the total reflection lens 3, the substrate 8 is spaced apart from the total reflection lens 3, and the substrate 8 is located outside the focal point of the objective lens 100. The micro-nanospheres 10 are sandwiched between the substrate 8 and the total reflection mirror 3. The sample is disposed on the surface of the substrate 8 facing the total reflection lens 3. The micro-nanospheres 10 are configured to control the spacing between the substrate 8 and the total reflection lens 3.
Referring to
A structure of the reflection-type objective lens in
In
Refer to the above description about
Referring to
The substrate 8 is made of a metal material or a non-metal material. If the substrate 8 is made of a metal material, a metal layer and a two-dimensional material film layer may not be provided on the surface of the substrate 8. If the substrate 8 is made of a non-metal material, optionally, a third metal layer 13 or a two-dimensional material film layer 14 is provided at a side of the substrate 8 facing the housing 1.
The material of the third metal layer 13 is selected from one or more of the group consisting of: gold, silver, copper, platinum, and palladium.
The material of the two-dimensional material film layer 14 is selected from one or more of the group consisting of: graphene, boron nitride, a transition metal chalcogenide, and a heterojunction composed of a transition metal chalcogenide.
The optical imaging device 750 provided in the above technical solution can be used in an optical sectioning microscope described later.
Referring to
The laser device 200 emits excitation light beam so as to excite samples. The laser emitted by the laser device 200 is single-wavelength light with a wavelength of, for example, 325 nm-1064 nm. The light beam is single-mode TEM 00, and the light beam energy distribution satisfies Gaussian equation. The light beam is linearly polarized, with a polarization ratio being not less than 30:1. A diameter of the light beam ranges from 1 mm-3 mm.
Referring to
The beam expanding rate of the beam expander 300 satisfies the following formula (8):
In the above, d is a light beam diameter after beam expanding, d′ is an initial light beam diameter, f is a focal length of a front lens 301, and f is a focal length of a rear lens 302.
The laser emitted by the laser device 200, after passing through the beam expander 300, becomes solid laser. The radius of the solid laser is expanded to the radius d by the beam expander 300. The excitation light beam with the expanded radius is converted by the beam changer 400 into an annular light beam with the radius r. The annular light beam means that a light spot obtained on a baffle or a detector which is arranged in front of the light beam is annular. The annular light beam is converted into an annular vector polarized light beam through the polarizer 500. The annular vector polarized light beam is incident into the objective lens 100 through a third reflecting mirror 550. After passing through the objective lens 100, vector light beam is focused in the sample region at the focal plane (160) in a conical shape according to an angle θ. The beam changer 400 adjusts r, and further adjusts an incident tapered angle θ. The beam expander 300 adjusts d, and further adjusts a coverage area Δθ of the incident tapered angle, i.e., a coverage angle adjustment area Δθ. According to different sample characteristics, the polarizer 500 is adjusted so as to select a suitable polarization state of excitation light, and the beam expander 300 and the beam changer 400 are adjusted so as to adjust the excitation angle θ and the excitation angle coverage area Δθ. The excitation angle θ can be adjusted within the range of 0˜85°, and the adjustment range is quite wide, therefore the applicable scenarios are greatly expanded, the coverage angle adjustment area Δθ is 0.5°˜5°, and the minimum adjustment amount can reach 1° or less, with quite high adjustment precision. The above technical solution has important application values in the field of optical spectra related to the excitation angle and the polarization state, such as the fields of plasmon-enhanced spectra and nano infrared spectra.
Referring to
The first axicon 401 and the second axicon 402 have the same refractive index, both being n. The tapered angles of the first axicon 401 and the second axicon 402 are equal, both being α. The solid light beam with the radius d, after passing through the beam changer 400, is converted into an annular light beam with the radius r. An edge thickness of the annular light beam is d″. r and d″ satisfy the following formula (9) and formula (10):
In the above, L is a minimum axial distance between the first axicon 401 and the second axicon 402 along an optical axis direction of each axicon; by driving the second axicon 402 to move along an electric slide rail 404 by the stepper motor 403, a position of the second axicon 402 can be adjusted, the magnitude of the distance L between the first axicon 401 and the second axicon 402 is further adjusted, and finally, the radius r of the annular light beam is adjusted.
Referring to
The wave plate has two main axes: a fast axis and a slow axis. Refractive indexes on the two axes are different, and therefore velocities of light thereof are also different. When the linearly polarized light is incident on the half-wave plate 502, and the polarization direction does not coincide with the fast axis or the slow axis, emergent light is still linearly polarized light, but rotates with respect to a polarization direction of the incident light. When circularly polarized light is incident on the half-wave plate 502, clockwise (counterclockwise) circularly polarized light will be converted into counterclockwise (clockwise) circularly polarized light.
The half-wave (λ/2) plate is configured to rotate a linear polarization direction of the incident light beam. When the half-wave plate 502 is mounted on a rotary mounting base, it can be used as a polarization optical rotator capable of continuous adjustment. In addition, when used in cooperation with a polarization beam splitter, the half-wave plate can be used as a beam splitter with a variable proportion. An included angle between a polarization direction of emergent light and a polarization direction of incident light is twice an included angle between a polarization direction of the incident light and a main axis of the wave plate. When the polarization direction of the incident light coincides with the fast axis or the slow axis, the polarization direction remains unchanged.
The distance between the linear polaroid 501, the half-wave plate 502, and the spiral wave plate 503 is set as required. By adjusting a rotation angle between the linear polaroid 501, the half-wave plate 502, and the spiral wave plate 503, the polarization state of the annular excitation light is adjusted.
The polarizer 500 converts the linearly polarized annular light beam into a radially or angularly polarized light beam, as the structure of the polarizer 500 shown in
To obtain linearly polarized light, the polarizer 500 satisfies the following relationships: the fast axis of the one-half wave plate is perpendicular to the fast axis of the spiral wave plate 503, and when an included angle between the fast axis of the half-wave plate 502 and the polarization direction of the linear polaroid 501 is β, an included angle between the polarization direction of emergent light and the linear polarization direction is 2β.
To obtain the radially polarized light, the polarizer 500 satisfies the following relationship: after passing through the one-half wave plate (i.e., the half-wave plate 502), the polarization direction of the linearly polarized light beam coincides with the fast axis of the spiral wave plate 503.
To obtain the angularly polarized light, the polarizer 500 satisfies the following relationship: after passing through the one-half wave plate (i.e., the half-wave plate 502), when the polarization direction of the linearly polarized light beam is perpendicular to the fast axis of the spiral wave plate 503, the emergent light is angularly polarized light.
With continued reference to
In some embodiments, the polarization direction of light output from the half-wave plate 502 coincides with the fast axis of the spiral wave plate 503, and the polarizer 500 outputs the radially polarized light.
In some embodiments, the polarization direction of light output from the half-wave plate 502 coincides with the fast axis of the spiral wave plate 503, and the polarizer 500 outputs the angularly polarized light.
Referring to
Referring to
Referring to
Referring to
Referring to
When it is necessary to reflect the light from the semi-reflective and semi-transmissive lens 605 into the objective lens 100, the second reflecting mirror 606 is located in the light path between the third reflecting mirror 550 and the objective lens 100. It can be understood that the light path is reversible, and when it is necessary to reversely propagate the light path in the direction described above, it is also required to move the second reflecting mirror 606 into the light path. When it is necessary to propagate light from the objective lens 100 into the spectrum detection device 700, the second reflecting mirror 606 is moved out of the light path between the third reflecting mirror 550 and the objective lens 100. The movement of the second reflecting mirror 606 is a linear reciprocating movement, which can be achieved by providing a linear reciprocating driving device, such as a linear motor. In order to improve the automation degree of the device and the precision of the movement control, a control device such as a displacement sensor or PLC may also be provided, so as to achieve precise control for the displacement of the second reflecting mirror 606.
A specific light path of the illumination device 600 during operation is described below: firstly, the second reflecting mirror 606 is moved into the light path, and subsequently, the illumination device 600 is adjusted to operate. Bandwidth visible light emitted from the visible light source 601 becomes parallel light after passing through the lens 602, then it is converted into annular light after passing through the annular baffle 603, and is changed in direction by the first reflecting mirror 604, after that, it is emitted to the semi-reflective and semi-transmissive lens 605, and then a part of light is irradiated to the movable second reflecting mirror 606, and enters the objective lens 100 of the optical imaging device 750. In the embodiment shown in
With continued reference to
The spectrum detection device 700 collects scattered light of the sample formed on the surface of the substrate 8, for subsequent optical detection and analysis. In this case, the second reflecting mirror 606 is located outside the light path. A collection sequence of light path of the spectrum detection device 700 is: the scattered light on the surface of the substrate 8, after passing through the objective lens 100, is transmitted to the third reflecting mirror 550, and a dichroscope 550a of the third reflecting mirror 550 transmits the light to an filter lens 703, and then the light is focused by the second focusing mirror 702, and finally enters the spectrometer 701. In the above, referring to
It should be noted first that, in the present embodiment and various embodiments below, basic steps of the optical detection method all include following several steps: adjusting the objective lens of the optical imaging device by the illumination device 600 so as to realize focusing; adjusting the polarization state, the excitation angle, and the coverage angle adjustment area of the laser light path of the imaging device 750 so as to make characteristics of the excitation light satisfy excitation requirements of the sample; and collecting an image of the sample under the laser excitation and analyzing the same. According to different specific detection requirements, manners of adjusting the excitation light are also different, and specific situations of various embodiments will be described in detail below. For parts which are not mentioned, refer to the detection process described in this paragraph, if without conflict.
With reference to
Step S1100: making a white light illumination device 600 operate until a clear image of nanoparticle or probe is observed on the imaging sensor 608.
In this step, the second reflecting mirror 606 is located in the light path. The light path of the illumination device 600 is as follows: bandwidth visible light emitted from the visible light source 601 of the illumination device 600 becomes parallel light after passing through the lens 602, then becomes annular light after passing through the annular baffle 603, after that, it passes through the first reflecting mirror 604, the semi-reflective and semi-transmissive lens 605, and the movable second reflecting mirror 606, and then enters the objective lens 100 of the optical imaging device 750. If it is the excitation nanoparticle 9, the light is focused near the nanoparticles 9. If it is the excitation probe, the light is focused near the tip of the probe. An inverted spectrometer structure is commonly used in the method of probe enhanced spectroscopy, and the excitation light polarization uses a radial polarization state. The light path of the illumination device 600 is likewise applicable to various embodiments below.
Taking the nanoparticles 9 as an example: light scattered by the nanoparticles 9 is collected by the objective lens 100, and subsequently passes through the second reflecting mirror 606 that is movable and the semi-reflective and semi-transmissive lens 605, then passes through the first focusing mirror 607, and finally, imaging is performed by the imaging lens on the surface of the imaging sensor 608. Whether or not the sample is located at the focal point of the objective lens 100 is judged by observing the imaging.
Step S1200: making the optical imaging device 750 and the spectrum detection device 700 operate, and in this case, the illumination device 600 no longer functioning. The laser emitted by the optical imaging device 750 will irradiate the region of the nanoparticles 9 or the region of the excitation enhanced spectral probe 12 at a high angle in a ring shape.
Step S1300: adjusting the angle coverage area of the excitation light. A beam expansion ratio is calculated according to a required radius of the laser light beam, and the beam expander 300 meeting the requirement of the beam expansion ratio is selected. For example, the radius of the laser light beam is 0.25 mm, and the beam expansion ratio of the beam expander 300 is 1.6, so that the output light beam has a radius of 0.4 mm.
Step S1400: adjusting an excitation angle of the excitation light. Specifically, by adjusting the position of the electric slide rail of the beam changer 400, the distance between the first axicon 401 and the second axicon 402 is changed, the radius of the annular light beam is adjusted, and further the excitation angle is adjusted.
Step S1500: determining a light polarization state of the excitation light, and adjusting the polarizer 500 to make the excitation light satisfy a required polarization state.
Specifically, according to properties and target characteristics of the sample 900 to be detected, the following configurations are making respectively:
An execution sequence of the above steps S1300, S1400, and S1500 is not limited.
Step S1600: focusing the collected sample signals from the region of nanoparticle or probe into the spectrometer 701.
In the above step S1200 to step S1600, after the polarization state, the excitation angle, and the coverage angle area are all determined, the optical imaging device 750 may operate, and the light path of the optical imaging device 750 during the operation is as follows: in this step, the second reflecting mirror 606 is located outside the light path. The diameter of the excitation light beam emitted by the laser device 200 is increased by the beam expander 300 to 2d. Subsequently, the beam changer 400 converts the light beam with the increased diameter into an annular light beam with a light beam wall thickness d and a light beam radius r. The polarizer 500 can control the polarization state of the annular light beam to switch between linear polarization and vector polarization, e.g., to switch to radial polarization. The third reflecting mirror 550 reflects the annularly polarized light beam to the objective lens 100, and then the light beam is focused on the surface of the substrate 8 at the excitation angle θ. In this case, the single nanoparticle 9 on the surface of the substrate 8 is located at the center of the focal point. In this case, the single nanoparticle 9 will enhance local electromagnetic field nearby, improve the excitation efficiency of sample molecules in the local field, and meanwhile amplify spectral signal of the molecules in the local field radiating to a far field. The spectral signal, after passing through the objective lens 100, is converted into a parallel light beam, which, after being transmitted through the third reflecting mirror 550, is filtered out the excitation light part thereof by the filter lens 703, and only the signal light part is left. Wavelengths of the signal light left, after being focused by the second focusing lens 702, enter the spectrometer 701 and are recorded.
Step S1700: analyzing a spectrogram collected by the spectrometer 701 to obtain chemical components of the sample.
In the detection method provided in the above technical solutions, the excitation angle θ and the excitation angle coverage area Δθ are respectively controlled by the beam expander 300 and the beam changer 400, and the excitation light polarization is controlled by the polarizer 500, therefore, the control over various vector polarized light fields such as any direction of linear polarization, angular polarization, and radial polarization at the focusing plane is realized.
With reference to
Before describing the detection method, an optical system on which the method is based will be described. The optical system on which the present detection method is based is substantially the same as the optical system used in
With reference to
Step S2100: making the white light illumination device 600 operate: moving the substrate 8 to below the objective lens 100, wherein when the substrate 8 is directly below the objective lens 100, a clear image of the surface of the sample can be observed on the surface of the substrate 8. For detailed description of the illumination device 600, reference is made to the above description, and it will not be repeated herein.
Step S2200: making the optical imaging device 750 and the spectrum detection device 700 operate, and in this case, the illumination device 600 no longer functioning. The laser emitted by the optical imaging device 750 will irradiate the region of the nanoparticles 9 or the region of the excitation enhanced spectral probe 12 at a high angle in a ring shape.
Step S2300: adjusting the angle coverage area of the laser. A beam expansion ratio is calculated according to a required radius of the laser light beam, and the beam expander 300 meeting the requirement of the beam expansion ratio is selected. For example, the radius of the laser light beam is 0.25 mm, and the beam expansion ratio of the beam expander 300 is 1.6, so that the output light beam has a radius of 0.4 mm.
Step S2400: adjusting an excitation angle of the laser. Specifically, by adjusting the position of the electric slide rail of the beam changer 400, the distance between the first axicon 401 and the second axicon 402 is changed, and the radius of the annular light beam is adjusted, further the excitation angle is adjusted, so that the excitation angle θ is larger than a total internal reflection critical angle.
The plane of the hemispherical prism coincides with the focal plane of the parabolic reflective objective lens 100, and the sphere center coincides with the focal point of the parabolic reflective lens. In this case, the excitation angle θ and the excitation angle coverage area Δθ are adjusted, so that the incident light is annularly focused at the region of the sphere center of the hemispherical prism. When excitation angle θ is greater than the total internal reflection critical angle, total reflection occurs in the region of the sphere center of the hemispherical prism, generating evanescent waves on an air side of the region of the sphere center. The nanoparticles 9 and the enhanced spectral probe in the evanescent wave range can enhance the local electromagnetic field intensity, and improve the excitation efficiency of sample molecules in the local field. The higher the excitation efficiency is, the more the energies of excitation light are coupled with the sample, thus improving the detection sensitivity of the device.
In some embodiments, in the detection method described in the present embodiment, the total reflection lens 3 (i.e. the structure described in
Step S2500, determining a polarization state of the laser, and adjusting the polarizer 500 to make the light satisfy a required polarization state.
Specifically, according to properties and target characteristics of the sample 900 to be detected, the following configurations are making respectively:
An execution sequence of the above steps S2300, S2400, and S2500 is not limited.
Step S2600: collecting sample signals from the region of particle or probe and then focusing the same into the spectrometer 701.
Step S2700, analyzing a spectrogram collected by the spectrometer 701 to obtain chemical components of the sample.
As can be seen from the detection process described above, the general process of optical detection is the same regardless of whether the total reflection lens 3 is provided, but the excitation angle and the polarization state may be different.
Referring to
In this detection method, a metal or two-dimensional material thin film that supports surface plasmon (i.e., SPP) is prepared on the surface of the substrate 8, and the sample is placed on the thin film. A lower surface of the substrate 8 is attached to the hemispherical prism through a refractive index coupling liquid. In this case, a clear image of the sample on the surface of the substrate 8 can be directly observed.
Referring to
step S3100: making a white light illumination device 600 operate until a clear image of the surface of the sample is observed on the surface of the substrate 8. For the detailed description of the illumination device 600, reference is made to the above description, and it will not be repeated herein.
Step S3200: making the optical imaging device 750 and the spectrum detection device 700 operate, and in this case, the illumination device 600 no longer functioning.
Step S3300: adjusting an excitation angle of the laser, so that the excitation angle reaches an SPP excitation angle. Specifically, by adjusting the position of the electric slide rail of the beam changer 400, the distance between the first axicon 401 and the second axicon 402 is changed, the radius of the annular light beam is adjusted, and the excitation angle is further adjusted, so that the excitation angle reaches an SPP excitation angle.
Step S3400: determining a polarization state of the laser, and adjusting the polarizer 500 to make the light satisfy a required polarization state.
In this mode, the sample responds optimally to polarization perpendicular to the surface of the substrate 8, therefore, radial polarization is used. In this case, the polarizer 500 is adjusted so that a linear polarization direction of the light beam passing through the one-half wave plate (i.e., the half-wave plate 502) coincides with the fast axis of the spiral wave plate 503.
An execution sequence of the above step S3300 and step S3400 is not limited.
Step S3500: collecting scattered light signals of the sample from the particle or probe region and then focusing the same into the spectrometer 701.
Step S3600: analyzing a spectrogram collected by the spectrometer 701 to obtain chemical components of the sample 900.
The above-described detection method is particularly suitable for detection of ultra-thin samples such as unimolecules and unimolecular layers.
Referring to
The Otto-SPP mode refers to a detection mode for implementing the optical detection principle of excited surface plasmon proposed by Otto. In the optical system on which this detection method is based, there is a distance between the substrate 8 thereof and the focal plane, i.e., reference can be made to the above structure described about
With reference to
Step S4200: making the optical imaging device 750 and the spectrum detection device 700 operate, and in this case, the illumination device 600 no longer functioning.
Step S4300: adjusting the angle coverage area of the laser. A beam expansion ratio is calculated according to a required radius of the laser light beam, and the beam expander 300 meeting the requirement of the beam expansion ratio is selected. For example, the radius of the laser light beam is 0.25 mm, and the beam expansion ratio of the beam expander 300 is 1.6, so that the output light beam has a radius of 0.4 mm.
Step S4400: adjusting an excitation angle of the laser. Specifically, by adjusting the position of the electric slide rail of the beam changer 400, the distance between the first axicon 401 and the second axicon 402 is changed, and the radius of the annular light beam is adjusted, and further the excitation angle is adjusted, so that the excitation angle reaches an SPP excitation angle.
Step S4500: determining a polarization state of the laser, and adjusting the polarizer 500 to make the light satisfy a required polarization state.
In this mode, the sample responds optimally to polarization perpendicular to the surface of the substrate 8, thus radial polarization is used. In this case, the polarizer 500 is adjusted so that a linear polarization direction of the light beam passing through the one-half wave plate (i.e., the half-wave plate 502) coincides with the fast axis of the spiral wave plate 503.
Step S4600: collecting sample signals from the region of nanoparticle 9 or excitation enhanced spectral probe 12 and then focusing the same into the spectrometer 701.
Step S4700: analyzing a spectrogram collected by the spectrometer 701 to obtain chemical components of the sample.
The optical detection method in the annular vector polarized excitation Otto-SPP mode is particularly suitable for detection of ultra-thin samples such as unimolecules and unimolecular layers. The spectrum detection has higher sensitivity, and can reach the detection sensitivity of single molecule.
Referring to
The optical system structure on which this detection mode is based will be described first.
Referring to
The first illumination device 600a has the same structure and function as the illumination device 600 described above with respect to the respective optical systems. The second illumination device 600b is of a structure substantially the same as that of the illumination device 600 described above, except that it is used to illuminate the microscopic objective lens 800. See the above for the structure and positional relationship between components of the first illumination device 600a, which will not be repeated herein. Only the second illumination device 600b is described.
Referring to
Referring to
With continued reference to
The optical detection method in the annular vector polarized excitation planar waveguide mode includes the following steps:
step S5100: enabling the first illumination device 600a and the second illumination device 600b until a clear image of the sample on the surface of the substrate 8 is directly observed.
Specific operation steps are: moving the second reflecting mirrors 606 and 606′ both into the light path, and adjusting the objective lens 100 and the microscopic objective lens 800, so that the surface of the substrate 8 can be seen clearly on both the imaging sensors 608 and 608′.
Step S5200: making the optical imaging device 750 and the spectrum detection device 700 operate, and in this case, the illumination device 600 no longer functioning.
Specifically, the second reflecting mirrors 606 and 606′ are both moved out of the light path, and in this case, the laser radiated from the laser device 200, after being expanded and shaped, is focused on the surface of the substrate 8 at a certain angle.
Step S5300: adjusting an excitation angle of the laser. Specifically, by adjusting a position of the electric slide rail of the beam changer 400, the distance between the first axicon 401 and the second axicon 402 is changed, the radius of the annular light beam is adjusted, and further the excitation angle is adjusted, so that the excitation angle reaches the waveguide mode excitation angle.
When the excitation light angle θ satisfies the excitation condition of the waveguide medium layer 62, an evanescent field is generated on the surface of the waveguide medium layer 62, and the local electromagnetic field of the nanoparticles 9 or the enhanced spectral probe located within the range of the evanescent field is further improved, further improving the excitation efficiency of the spectrum.
Step S5400: determining a polarization state of the laser, and adjusting the polarizer 500 to make the laser satisfy a required polarization state.
Specifically, according to properties and target characteristics of the sample 900 to be detected, the following configurations are made respectively:
Through the adjustments in step S5300 and step S5400, the excitation angle and the polarization state of the laser are further enabled to meet the waveguide excitation condition.
Step S5500: collecting sample signals from the region of nanoparticle 9 or excitation enhanced spectral probe 12 and then focusing the same into the spectrometer 701. Spectral signals originated from the substrate 8 are recorded by the spectrometer 701 after passing through the objective lens 100 and subsequent light path. Meanwhile, another other part of spectral signals originated from the substrate 8 are recorded by the spectrometer 701′ after passing through the microscopic objective lens 800 and subsequent light path.
Step S5600: analyzing a spectrogram collected by the spectrometers 701 and 701′ to obtain chemical components of the sample.
In the above optical detection method, the amount of samples may be very small, more excitation angles are supported, and more polarization modes are supported, therefore, it has wider application scenarios.
Referring to
In this mode, the optical system includes the illumination device 600 and the optical imaging device 750. Referring to
How to use the optical system provided in the present embodiment to realize the optical detection is described below.
Step S6100: making the white light illumination device 600 operate, i.e., the illumination device 600 functioning: moving the substrate 8 to below the objective lens 100, wherein when the substrate 8 is directly below the objective lens 100, a clear image of the surface of the sample can be observed on the surface of the substrate 8. Specific operation steps are as follows: the second reflecting mirror 606 is moved into the light path, the visible light source 601 emits broadband white light, which is changed into parallel light after passing through the lens 602, then changed into annular light after passing through the annular baffle 603, after that, it is reflected by the first reflecting mirror 604, then reflected by the second reflecting mirror 606 and the third reflecting mirror 550, entering the objective lens 100, and finally forming a light spot on the substrate 8. The position of the sample or the objective lens 100 is moved until the sample 900 is seen clearly.
Step S6200: adjusting the optical system so that the optical imaging device 750 and the spectrum detection device 700 both operate, in this case, the illumination device 600 no longer functioning. In this case, the second reflecting mirror 606 is moved out of the light path.
Step S6300: adjusting an excitation angle. Specifically, by adjusting a position of the electric slide rail of the beam changer 400, the distance between the first axicon 401 and the second axicon 402 is changed, the radius of the annular light beam is adjusted, and further the excitation angle is adjusted, so that the excitation angle reaches the maximum excitation angle tolerable by the objective lens 100.
Step S6400: using the optical imaging device 750 for illumination, i.e., laser illumination.
Step S6500: adjusting and controlling the polarization state of the laser to switch between the radial polarization and the angular polarization, so as to obtain fluorescent imaging of the sample in each polarization state.
Specifically, according to properties and target characteristics of the sample 900 to be detected, the following configurations are made respectively:
Step S6600: comparing imaging of the sample 900 in different polarization states, and analyzing an internal structure of the sample 900.
Referring to
Reference is made to the description of
In this mode, there is a further set of optical imaging device 750 in the optical system on the basis of the optical system described above with reference to
Referring to
How to use the optical system provided in the present embodiment to implement optical detection is introduced below.
Step S7100: making the white light illumination device 600 operate, i.e., the illumination device 600 functioning: moving the substrate 8 to below the objective lens 100, wherein when the substrate 8 is directly below the objective lens 100, a clear image of the surface of the sample can be observed on the surface of the substrate 8. For the illumination device 600, specific operation steps are as follows: moving the second reflecting mirror 606 into the light path, the visible light source 601 emits broadband white light, which is changed into parallel light after passing through the lens 602, then changed into annular light after passing through the annular baffle 603, after that, it is reflected by the first reflecting mirror 604, then reflected by the second reflecting mirror 606 and the third reflecting mirror 550, and entering the objective lens 100. The position of the sample or the objective lens 100 is moved until the sample is seen clearly.
Step S7200: making the two optical imaging devices 750 and the spectrum detection device 700 both operate, and in this case, the illumination device 600 no longer functioning. In this case, the second reflecting mirror 606 is moved out of the light path.
Step S7300: adjusting excitation angles of respective lasers. Specifically, by adjusting the position of the electric slide rail of the beam changer 400, the distance between the first axicon 401 and the second axicon 402 is changed, the radius of the annular light beam is adjusted, and further the excitation angle is adjusted, so that the excitation angle reaches the maximum excitation angle tolerable by the objective lens 100.
Step S7400: using two paths of laser to illuminate. Relative positions of the two paths of laser are adjusted so that the light path difference is 0 when the two paths of light are combined at the beam combiner 1000.
Step S7500: adjusting and controlling the polarization state to switch between the radial polarization and the angular polarization, to obtain fluorescent imaging of the sample in each polarization state.
Specifically, according to properties and target characteristics of the sample 900 to be detected, the following configurations are made respectively:
Specific operations of step S7300-step S7500 are as follows: at this time, the second reflecting mirror 660 has been moved out of the light path. The two laser devices 200 and 200′ respectively emit laser. The two paths of laser, after being adjusted in the light beam and the polarization state, are combined at the beam combiner 1000, and the combined laser is reflected by the third reflecting mirror 550 and enters the objective lens 100, then it is focused on the substrate 8. A delayer 2000 is adjusted, a light path difference between a left optical arm and a lower optical arm at the beam combiner 1000 is moved, wherein when the light path difference between the two is 0, two-photon coupling will be generated in the sample at the substrate 8, and fluorescence photons radiated from the sample, after passing through the objective lens 100, the dichroscope 550a, and the fluorescence filter 609, are focused on the imaging sensor 608 by an imaging lens 607. In the above imaging process, by performing the adjustments in step S7300-step S7500, different imaging effects can be obtained.
Step S7600: comparing imaging of the sample 900 in different polarization states, and analyzing an internal structure of the sample 900.
The optical detection in the annular vector polarized excitation optical sectioning microscope mode mainly aims at life science samples such as cells and tissues.
The optical system provided in various technical solutions above limits the energy of excitation light within a range of 1-degree excitation angle, and significantly improves the light energy excitation efficiency; moreover, the excitation angle is adjustable in 0-80 degrees, which fully satisfies the requirements of the samples on the excitation angle; the polarization state is flexibly adjustable, which fully satisfies the requirements of the samples on the excitation polarization; moreover, this optical system realizes that the angle control capability achieved by the conventional large and complex corner device is reduced to the integrated optical imaging device 750, simplifies the structure of the device, and can be combined with existing microscopic spectrum systems or develop portable spectrum devices.
In the description of the present disclosure, it should be understood that orientation or positional relations indicated by terms such as “center”, “longitudinal”, “transverse”, “front”, “rear”, “left”, “right”, “vertical”, “horizontal”, “top”, “bottom”, “inner”, and “outer” are based on orientation or positional relations as shown in the accompanying drawings, merely for facilitating the description of the present disclosure and simplifying the description, rather than indicating or implying that related devices or elements have to be in the specific orientation or be configured and operated in a specific orientation, therefore, they should not be construed as limitation on the scope of protection of the present disclosure.
Finally, it should be explained that the above embodiments are merely used for illustrating the technical solutions of the present disclosure, rather than limiting the present disclosure; although the detailed description is made to the present disclosure with reference to preferred embodiments, those ordinarily skilled in the art should understand that they still could modify the embodiments of the present disclosure or make equivalent substitutions to some of the technical features therein; and these modifications or substitutions, without departing from the spirit of the technical solution of the present disclosure, should be all covered within the scope of the technical solutions claimed in the present disclosure.
Number | Date | Country | Kind |
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202010879955.4 | Aug 2020 | CN | national |
Filing Document | Filing Date | Country | Kind |
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PCT/CN2021/108972 | 7/28/2021 | WO |