Adjustable three-dimensional fluorescence spectrum system

Abstract

The invention discloses an adjustable three-dimensional fluorescence spectrum system, which comprises a broad spectrum light source, a pair of linear gradual change optical filters F1 and F2, light collimators or lenses, a sample cavity, an imaging device, a structural member and a power supply control circuit, the spectrum directions of the F1 and the F2 intersect, the light collimators or lenses are arranged between the light source and the F1 and between the imaging device and the F2, and the structural member and the control circuit can control the angles of the F1 and the F2; the data acquisition and adjustment mode is that the horizontal direction x of an image output by an imaging device is consistent with the spectral direction of F1, the vertical direction y is consistent with the spectral direction of F2, the diagonal line is an absorption spectrum, the upper and lower half matrixes are three-dimensional fluorescence spectrums, and the included angle between F1 and F2 is smaller than or larger than 90 degrees through rotation so as to adjust the resolution of an emission spectrum or a receiving spectrum. The system solves the problems that an existing device is complex in structure, high in cost, long in scanning time and difficult in rapid detection, and flexible adjustment and efficient detection are achieved.

Description

Technical Field

[0001] This invention provides a spectroscopic system, and particularly relates to an adjustable three-dimensional fluorescence spectroscopic system. Background Technology

[0002] Three-dimensional fluorescence spectroscopy is a matrix spectrum characterized by three-dimensional coordinates of excitation wavelength (y-axis), emission wavelength (x-axis), and fluorescence intensity (z-axis), also called total emission spectrum. Its function is to acquire fluorescence emission spectra at different excitation wavelengths to form three-dimensional projection maps or contour maps, thereby enabling comprehensive analysis of the fluorescence characteristics of a sample. Currently, there are three main types of devices for acquiring three-dimensional fluorescence spectra: first, general fluorescence spectrophotometers, which require separate measurement of emission spectra at different excitation wavelengths, making operation cumbersome; second, microcomputer-controlled automatic scanning and recording fluorescence spectrophotometers, which repeatedly scan the emission spectrum and process data under fixed excitation wavelength increments via microcomputer control, resulting in a relatively complex structure; and third, television fluorometers, composed of orthogonal polychromators, television detectors, and computer interfaces, which can automatically acquire three-dimensional fluorescence spectra, but are more expensive.

[0003] Existing three-dimensional fluorescence spectrometers have significant shortcomings in terms of structure and performance: on the one hand, their emission spectral scanning structures are complex, such as television fluorometers which contain a variety of precision components, resulting in high manufacturing costs and making them difficult to widely adopt; on the other hand, their scanning time is long, whether it is a manually operated general fluorescence spectrophotometer or an automatically scanning device, they all need to perform spectral scanning point by point or segment by segment, which cannot achieve rapid detection and is difficult to meet the needs of scenarios with high detection efficiency requirements. Summary of the Invention

[0004] To address the aforementioned issues, this application provides an adjustable three-dimensional fluorescence spectroscopy system, which solves the problems of complex structure, high cost, long scanning time, and difficulty in rapid detection in existing devices, thereby achieving flexible adjustment and efficient detection.

[0005] To solve the above-mentioned technical problems, the present invention provides the following technical solution: an adjustable three-dimensional fluorescence spectroscopy system, including a broadband light source, a pair of linear gradient filters F1 and F2, an optical collimator or lens, a sample cavity, an imaging device, structural components and a power supply and control circuit;

[0006] The spectral directions of the pair of linear graded filters F1 and F2 intersect.

[0007] A collimator or lens is provided between the light source and the linear gradient filter F1 to uniformly project light into the sample cavity;

[0008] A collimator or a lens is provided between the imaging device and the linear variable filter F2, which is used to project the light absorbed by the sample in the sample chamber or emitted by fluorescence excitation of the sample onto the pixels at corresponding positions on the surface of the imaging device one by one. The image collected by the imaging device corresponds to the spectral data of the sample.

[0009] The structural member is used for fixing and moving each component. The power supply control circuit is used to control the operation of the system, and the structural member and the control circuit can control the angles of the linear variable filters F1 and F2.

[0010] The data acquisition and adjustable mode of this system is as follows:

[0011] (1) The horizontal direction x of the image output by the imaging device is consistent with the spectral direction of F1, and the vertical direction y is consistent with the spectral direction of F2. The diagonal direction of the image output by the imaging device is the absorption spectrum of the sample, and the upper half matrix and the lower half matrix are three-dimensional fluorescence spectra.

[0012] (2) By rotation, the included angle between the spectral directions of the linear variable filters F1 and F2 is made less than 90° or greater than 90° to correspondingly adjust the spectral resolution of the emission spectrum or the reception spectrum.

[0013] Preferably: The broad-spectrum light source is composed of a small number of LEDs with different wavelengths and photosensitive elements attached with different filters, or is composed of a xenon lamp and a rotatable filter with a specific wavelength.

[0014] Preferably: The broad-spectrum light source is a xenon lamp or an LED array.

[0015] Preferably: The spectral directions of the pair of linear variable filters F1 and F2 are perpendicular.

[0016] Preferably: The linear variable filter is made by an ion-assisted method or an ion beam sputtering method.

[0017] Preferably: The imaging device is a CCD or a CMOS camera.

[0018] Preferably: The layout of the collimator and the lens in the optical path is optimized to reduce stray light.

[0019] Preferably: The system further includes an efficient spectral data processing algorithm for realizing spectral extraction and analysis.

[0020] Preferably: The horizontal direction x corresponds to the spectral distribution of the incident light, and the vertical direction y corresponds to the spectral distribution of the fluorescence or transmitted light emitted by the sample; the upper half matrix (y>x) reflects the fluorescence emission characteristics of the sample at different excitation wavelengths, and the lower half matrix (y<x) reflects the non-fluorescent process.

[0021] Preferably, when the angle between the spectral directions of linear gradient filters F1 and F2 is less than 90°, the resolution of the emission spectrum is improved; when the angle is greater than 90°, the resolution of the reception spectrum is improved.

[0022] The change in light intensity along the diagonal direction (x = y) reflects the absorption characteristics of the sample for different wavelengths of light.

[0023] One or more technical solutions provided in the embodiments of this application have at least the following technical effects or advantages compared with the prior art:

[0024] Existing three-dimensional fluorescence spectroscopy designs suffer from problems such as complex emission spectral scanning structures, high costs, long scanning times, and difficulties in rapid detection. This invention addresses these issues by providing an adjustable three-dimensional fluorescence spectroscopy system. This system includes a broad-spectrum light source, a pair of linear graded filters, an optical collimator or lens, a sample chamber, an imaging device, necessary structural components, and a power supply and control circuit. It employs a small number of LEDs of different wavelengths and photosensitive elements with different filters attached. Utilizing a pair of linear graded filters with intersecting (preferably perpendicular) spectral directions, the light source and the linear graded filter F1... Optical path optimization is achieved through optical collimators or lenses between the imaging device and the linear graded filter F2, ensuring that the horizontal x-direction of the output image from the imaging device is aligned with the spectral direction of F1, and the vertical y-direction is aligned with the spectral direction of F2. This results in the absorption spectrum of the sample along the diagonal, and the three-dimensional fluorescence spectrum along the upper and lower half-matrix. Simultaneously, the spectral resolution of the emission or reception spectrum can be adjusted by rotating the linear graded filters F1 and F2 so that the angle between their spectral directions is less than 90° or greater than 90°. This achieves the effects of low system design cost, simple structure, fast detection speed, and flexible adjustment method.

[0025] Other advantages, objectives and features of the invention will be set forth in part in the description which follows, and in part will be apparent to those skilled in the art from the following examination or study, or may be learned from the practice of the invention. Attached Figure Description

[0026] Figure 1 This is a schematic diagram of the installation of an adjustable three-dimensional fluorescence spectroscopy system according to the present invention;

[0027] Figure 2 This is a schematic diagram of the broad-spectrum light source structure of an adjustable three-dimensional fluorescence spectroscopy system according to the present invention;

[0028] Figure 3 This is a schematic diagram of the structure and spectral direction of a linear gradient filter (F1 / F2) for an adjustable three-dimensional fluorescence spectroscopy system according to the present invention;

[0029] Figure 4This is a schematic diagram of the spectral information distribution of the output image of the imaging device of the adjustable three-dimensional fluorescence spectroscopy system of the present invention;

[0030] Figure 5 This is a schematic diagram illustrating the relationship between the angle adjustment of the linear gradient filter and the spectral resolution of an adjustable three-dimensional fluorescence spectroscopy system according to the present invention. Detailed Implementation

[0031] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0032] It should be noted that the terms "vertical," "horizontal," "up," "down," "left," "right," and similar expressions used in this article are for illustrative purposes only and do not represent the only possible implementation.

[0033] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains; the terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to limit the invention; the term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0034] like Figure 1 As shown, an adjustable three-dimensional fluorescence spectroscopy system includes a broadband light source, a pair of linear gradient filters, a sample chamber, an imaging device and structural components, and a power supply and control circuit.

[0035] The resolution of emission and fluorescence spectra is achieved by using a small number of LEDs of different wavelengths and photosensitive elements with different filters to acquire data of specific fluorescence spectra. The system has low cost, simple structure and fast detection speed.

[0036] The spectral directions of a pair of linear graded filters F1 and F2 intersect, preferably perpendicularly. A collimator or lens can be added between the light source and the linear graded filter F1 to uniformly project light into the sample cavity. A collimator or lens can be added between the imaging device and the linear graded filter F2 to project the light absorbed by the sample or emitted by fluorescence excitation in the sample cavity onto the corresponding pixels on the surface of the imaging device. The image acquired by the imaging device corresponds to the spectral data of the sample.

[0037] This system can adjust the spectral resolution of the emission or reception spectrum by changing the angle between the linear graded filters F1 and F2. The adjustment method is as follows:

[0038] (1) The horizontal x direction of the image output by the imaging device is consistent with the F1 spectral direction, and the vertical y direction is consistent with the F2 spectral direction. Then the diagonal direction of the image output by the imaging device is the absorption spectrum of the sample, and the upper half matrix and the lower half matrix are the three-dimensional fluorescence spectrum.

[0039] (2) By rotating, the spectral direction angle between linear gradient filters F1 and F2 can be made less than 90° or greater than 90°, so the spectral resolution of the emission spectrum or the receiving spectrum can be adjusted accordingly.

[0040] In this embodiment, a broadband light source provides light with a wide wavelength range. After being collimated by the light collimator or lens between the light source and F1, the light is uniformly projected onto F1. F1 modulates the incident light spectrally and then illuminates the sample in the sample cavity. The sample absorbs light or generates fluorescence, which is separated by F2. The fluorescence is then precisely projected onto the corresponding pixel of the imaging device through the light collimator or lens between F2 and the imaging device. The structural components fix each part and work with the power supply control circuit to achieve the angle adjustment of F1 and F2. From the perspective of implementation key points and innovation, the light source composed of a small number of LEDs of different wavelengths and photosensitive elements with different filters simplifies the structure and reduces costs. It can achieve the acquisition of specific fluorescence spectrum data without complex scanning mechanisms, which greatly improves the detection speed. The spectral directions of F1 and F2 intersect (preferably perpendicular), and combined with the image distribution characteristics of the imaging device, the diagonal corresponds to the absorption spectrum and the upper and lower half of the matrix corresponds to the three-dimensional fluorescence spectrum, realizing the synchronous acquisition of multispectral information. The design of adjusting the resolution by rotating F1 and F2 to change the included angle allows the system to flexibly optimize the accuracy of the emission spectrum or the receiving spectrum according to the needs. This solves the problems of complex structure, high cost, slow scanning and limited adjustment in the existing technology. The synergistic effect of each structure significantly improves the practicality and economy of the system while ensuring the detection effect.

[0041] It should be noted that, for example Figure 2 and Figure 3 As shown, (1) Broad spectrum light source: The light source designed in this system is a broad spectrum light source, which is designed to consist of a small number of LEDs of different wavelengths and photosensitive elements with different filters attached, or a xenon lamp combined with a rotatable filter of a specific wavelength; providing light with a wide wavelength range to excite the sample to produce fluorescence or absorption spectrum.

[0042] Preferably, this system selects a light source with high stability and a wide spectral range, such as a xenon lamp or an LED array, to ensure the uniformity and coverage of the excitation light.

[0043] (2) Linear graded filters F1 and F2: This system uses a pair of linear graded filters F1 and F2 as spectroscopic elements. F1 is used to modulate the spectral distribution of the incident light, and F2 is used to separate the fluorescence or transmitted light emitted by the sample. The spectral directions of this system intersect, preferably perpendicularly; through the intersection of the spectral directions, preferably perpendicularly, spatial separation of light of different wavelengths is achieved.

[0044] Preferably, the system uses a high-quality linear graded filter to ensure the accuracy and efficiency of spectral separation. A linear graded filter is an optical device whose spectral characteristics change linearly with position. It is formed by depositing multiple layers of films with varying thicknesses on a substrate surface using processes such as ion-assisted methods or ion beam sputtering. Compared to traditional narrowband filters, linear graded filters have near-continuous spectral channels, thus achieving higher spectral resolution through spectral dispersion. Compared to prism and grating-type spectral imagers, spectral imagers based on linear graded filters offer higher integration, higher stability, and higher resolution. They also feature a compact structure, small size, light weight, and lower research and manufacturing costs.

[0045] (3) Sample chamber: Place the sample to be tested, and the light source illuminates the sample through F1. After the sample absorbs or excites fluorescence, the light is received by the imaging device through F2.

[0046] (4) Imaging device: The image acquired by the imaging device in this system corresponds to the spectral data of the sample, converting the spectral information into a spatially distributed image. Each pixel in the image corresponds to the light intensity information of a specific wavelength.

[0047] Preferably, the imaging device is a high-sensitivity, low-noise imaging device such as a CCD or CMOS camera to improve the signal-to-noise ratio of the spectral data.

[0048] (5) Optical collimator or lens: to ensure the uniformity and efficiency of the optical path and improve the signal-to-noise ratio and resolution of the system.

[0049] A collimator or lens can be added between the light source and the linear gradient filter F1 to project light evenly into the sample cavity;

[0050] A collimator or lens can be added between the imaging device and the linear gradient filter F2, so that the light absorbed by the sample in the sample cavity or emitted by fluorescence can be projected one-to-one onto the corresponding pixels on the surface of the imaging device.

[0051] Preferably, the layout of the optical collimator and lens can be optimized in the optical path design to reduce stray light in the optical path and improve the optical efficiency of the system.

[0052] (6) Structural components and power supply control circuit: The structural components are used for fixing and moving each part; the control circuit part controls the operation of the system. The structural components and control circuit can control the angles of the linear gradient filters F1 and F2; the system develops an efficient spectral data processing algorithm in the software algorithm to achieve fast and accurate spectral extraction and analysis.

[0053] When using, such as Figure 1 , 4 As shown in Figure 5, the following precautions should be taken when adjusting this system:

[0054] (1) The horizontal x direction of the image output by the imaging device is consistent with the F1 spectral direction, and the vertical y direction is consistent with the F2 spectral direction. Then the diagonal direction of the image output by the imaging device is the absorption spectrum of the sample, and the upper half matrix and the lower half matrix are the three-dimensional fluorescence spectrum.

[0055] (2) By rotating, the spectral direction angle between linear gradient filters F1 and F2 can be made less than 90° or greater than 90°, so the spectral resolution of the emission spectrum or the receiving spectrum can be adjusted accordingly.

[0056] In this system, the spectral data is extracted as follows: the horizontal direction (x-axis) is consistent with the spectral direction of F1, corresponding to the spectral distribution of the incident light; the vertical direction (y-axis) is consistent with the spectral direction of F2, corresponding to the spectral distribution of the fluorescence or transmitted light emitted by the sample; the diagonal direction reflects the absorption spectrum of the sample; the upper half matrix and the lower half matrix correspond to the excitation and emission information of the three-dimensional fluorescence spectrum, respectively.

[0057] In this system, the spectral resolution is adjusted by rotating F1 and F2, changing the angle between their spectral directions (less than 90° or greater than 90°). An angle less than 90° improves the resolution of the emission spectrum, suitable for scenarios requiring high-precision fluorescence analysis. An angle greater than 90° improves the resolution of the received spectrum, suitable for scenarios requiring high-precision absorption spectral analysis.

[0058] (1) In this system, the diagonal direction reflects the absorption spectrum of the sample:

[0059] In the two-dimensional image output by the imaging device, the diagonal direction (i.e., the direction where x = y) indicates that the incident light wavelength (x-axis) and the emitted light wavelength (y-axis) are the same. When light passes through the sample, it is neither absorbed nor undergoes wavelength shift (e.g., no fluorescence emission occurs). Therefore, the change in light intensity along the diagonal direction directly reflects the sample's absorption characteristics for different wavelengths of light. When incident light passes through the sample, the sample absorbs light of specific wavelengths. The intensity of the absorbed light is related to the sample's absorption characteristics and follows Beer-Lambert's law:

[0060] I(λ)=I0(λ)·e -α(λ)·c·d

[0061] Where: I(λ) is the intensity of the emitted light; I0(λ) is the intensity of the incident light; α(λ) is the absorption coefficient of the sample; c is the sample concentration; d is the optical path length. In the diagonal direction (x = y), the ratio of the intensity of the emitted light I(λ) to the intensity of the incident light I0(λ) reflects the absorption characteristics of the sample:

[0062]

[0063] By measuring this ratio, the absorption spectrum of the sample can be obtained. If the sample absorbs light strongly at a certain wavelength, the light intensity at the diagonal position corresponding to that wavelength is low. If the sample absorbs light weakly at a certain wavelength, the light intensity at the diagonal position corresponding to that wavelength is high. The absorption spectrum of the sample can be plotted based on the light intensity distribution in the diagonal direction.

[0064] This design utilizes the characteristics of the linear variable filter and the imaging device to achieve the simultaneous measurement of the absorption spectrum and the fluorescence spectrum, with high experimental efficiency and flexibility.

[0065] (2) In this system, the upper half matrix and the lower half matrix respectively correspond to the excitation and emission information of the three-dimensional fluorescence spectrum.

[0066] Upper half matrix (y > x): Directly corresponds to the three-dimensional fluorescence spectrum, reflecting the fluorescence emission characteristics of the sample at different excitation wavelengths;

[0067] Lower half matrix (y < x): Mainly reflects non-fluorescence processes, such as scattering or Raman scattering.

[0068] The fluorescence spectrum usually includes two key parts: the excitation spectrum, which describes the fluorescence emission intensity of the sample at different excitation wavelengths; the emission spectrum, which describes the wavelength distribution of the fluorescence emitted by the sample at a specific excitation wavelength. In the three-dimensional fluorescence spectrum, the excitation wavelength x-axis, the emission wavelength y-axis, and the fluorescence intensity z-axis together form a three-dimensional data space. In the diagonal direction (x = y) of the two-dimensional image output by the imaging device in this system: it reflects the absorption spectrum of the sample (the incident light and the emitted light have the same wavelength), and the non-diagonal direction (x ≠ y) reflects the fluorescence spectrum of the sample (the incident light and the emitted light have different wavelengths). According to the relationship between x and y, the image can be divided into:

[0069] Upper half matrix (y > x), the wavelength of the emitted light (y-axis) is greater than the wavelength of the incident light (x-axis): This corresponds to fluorescence emission because the wavelength of fluorescence emission is usually longer than the excitation wavelength (Stokes shift). The upper half matrix reflects the fluorescence emission characteristics of the sample at different excitation wavelengths.

[0070] Lower half matrix (y < x), where the wavelength of the outgoing light (y-axis) is less than the wavelength of the incident light (x-axis): This usually corresponds to non-fluorescent processes such as scattering or Raman scattering. The lower half matrix may contain minor information of the fluorescence spectrum in some cases, but mainly reflects non-fluorescent characteristics.

[0071] The upper half matrix (y > x) directly corresponds to the excitation-emission matrix of the three-dimensional fluorescence spectrum: x-axis, excitation wavelength (determined by F1); y-axis, emission wavelength (determined by F2); light intensity (z-axis), fluorescence intensity. By analyzing the upper half matrix, the following information of the sample can be extracted: Optimal excitation wavelength: The excitation wavelength with the maximum fluorescence intensity; Optimal emission wavelength: The emission wavelength with the maximum fluorescence intensity; Shape and position of the fluorescence peak: Reflect the fluorescence characteristics of the sample.

[0072] The lower half matrix (y < x) mainly reflects non-fluorescent processes, but in some cases, it may also contain the following information: Raman scattering: The wavelength of the outgoing light is less than the wavelength of the incident light; Second harmonic generation: Signals generated by nonlinear optical effects; System noise or stray light: Needs to be removed through data processing.

[0073] This system can extract the three-dimensional fluorescence spectrum information of the sample by analyzing the upper half matrix, including the excitation spectrum, emission spectrum and fluorescence intensity distribution. This design cleverly utilizes the characteristics of the linear variable filter and the imaging device to achieve efficient and flexible spectral measurement.

[0074] (3) In this system, by changing the spectral direction angle (less than 90° or greater than 90°) between the linear variable filters F1 and F2, the spectral resolution of the emission spectrum or the reception spectrum can be adjusted.

[0075] A linear variable filter is an optical element whose transmission wavelength changes linearly with spatial position. For example, for F1, the spectral direction is along the x-axis, and the transmission wavelength increases gradually from left to right. For F2, the spectral direction is along the y-axis, and the transmission wavelength increases gradually from bottom to top. When the spectral direction angle between F1 and F2 is 90°, the directions of their transmission wavelength changes are perpendicular to each other, forming a two-dimensional spectral separation plane.

[0076] Spectral resolution refers to the ability of the system to distinguish two adjacent wavelengths, which is mainly affected by the following factors: Spectral gradient of the filter: The rate of change of the filter transmission wavelength with spatial position; Spatial resolution of the imaging device: The minimum spatial distance that the imaging device can resolve; Optical path design: The angle between the filters and the collimation of the optical path.

[0077] By rotating F1 and F2 to change their spectral direction angle (less than 90° or greater than 90°), the spectral resolution can be adjusted. The specific working principle is as follows:

[0078] When the angle is less than 90°, the spectral directions are closer to parallel, and the transmission wavelength changes of F1 and F2 are more consistent. In imaging devices, light of the same wavelength is distributed into a smaller spatial region. This improves the resolution of the system's emission spectrum because the wavelength distribution of the emitted light is more finely separated. However, it reduces the resolution of the received spectrum because the wavelength distribution of the incident light is compressed.

[0079] When the angle is greater than 90°, the spectral direction is closer to perpendicular, and the transmission wavelength changes of F1 and F2 are more orthogonal. In imaging devices, light of the same wavelength is distributed over a larger spatial area. This improves the resolution of the received spectrum because the wavelength distribution of the incident light is more finely separated. Conversely, it reduces the resolution of the emitted spectrum because the wavelength distribution of the emitted light is broadened.

[0080] Assume: the spectral gradient of F1 is G1 (unit: nm / mm), the spectral gradient of F2 is G2 (unit: nm / mm), and the included angle is θ. On the imaging device, the relationship between spectral resolution Δλ and spatial resolution Δx is:

[0081]

[0082] When θ < 90°, cosθ > 0, and the spectral resolution is mainly determined by the emission spectrum; when θ > 90°, cosθ < 0, and the spectral resolution is mainly determined by the reception spectrum. By adjusting the angle between F1 and F2, the system's measurement of different spectral characteristics can be optimized: for high emission spectral resolution, setting the angle to less than 90° is suitable for fluorescence spectral measurements, especially for scenarios requiring the differentiation of similar fluorescence peaks. For high reception spectral resolution, setting the angle to greater than 90° is suitable for absorption spectral measurements, especially for scenarios requiring the differentiation of similar absorption peaks.

[0083] This system allows for adjustment of spectral resolution by changing the angle between the spectral directions of F1 and F2. This design provides flexible spectral resolution adjustment capabilities, enabling the system to adapt to different experimental needs.

[0084] It should be noted that during the use of this device, the following additional components are required: a sample cell (such as a quartz cuvette, used to hold liquid samples and placed inside the sample chamber), a heat dissipation device (such as an aluminum heat sink, installed at the LED or xenon lamp of the broad-spectrum light source to avoid overheating and affecting the stability of the light source), a data transmission line (such as a USB or gigabit network cable, connecting the imaging device and the computer to realize image data transmission), a computer equipped with spectral analysis software (used to process the image data output by the imaging device and convert it into a three-dimensional fluorescence spectrum), an optical platform (such as a cast iron platform, used to fix various structural components to ensure the stability of the optical path), and a calibration standard sample (such as Rhodamine B solution, used to calibrate the spectral accuracy of the system). Among these, the substrate of the linear gradient filters F1 and F2 can be made of quartz material, the coating layer is formed by alternating stacking of silicon dioxide and magnesium fluoride, the sample chamber can be made of 304 stainless steel, the lens of the collimator can be made of K9 optical glass, and the shell of the imaging device can be made of 6061 aluminum alloy to ensure the compatibility of each component and the overall stability of the system.

[0085] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. An adjustable three-dimensional fluorescence spectroscopy system, characterized in that, It includes a broad-spectrum light source, a pair of linear gradient filters F1 and F2, a collimator or lens, a sample chamber, an imaging device, a structural member, and a power supply control circuit; The spectral directions of the pair of linear gradient filters F1 and F2 intersect; A collimator or lens is provided between the light source and the linear gradient filter F1 for uniformly projecting light into the sample chamber; A collimator or lens is provided between the imaging device and the linear gradient filter F2 for projecting the light absorbed by the sample in the sample chamber or emitted by fluorescence excitation onto the pixels at corresponding positions on the surface of the imaging device one by one, and the image collected by the imaging device corresponds to the spectral data of the sample; The structural member is used for fixing and moving each component, and the power supply control circuit is used for controlling the system operation, and the structural member and the control circuit can control the angles of the linear gradient filters F1 and F2; The data acquisition and adjustable mode of this system is as follows: (1) The horizontal direction x of the image output by the imaging device is consistent with the spectral direction of F1, and the vertical direction y is consistent with the spectral direction of F2. The diagonal direction of the image output by the imaging device is the absorption spectrum of the sample, and the upper half matrix and the lower half matrix are three-dimensional fluorescence spectra; (2) By rotating, the included angle between the spectral directions of the linear gradient filters F1 and F2 is made less than 90° or greater than 90° to correspondingly adjust the spectral resolution of the emission spectrum or the reception spectrum.

2. The system according to claim 1, characterized in that, The broad-spectrum light source is composed of a small number of LEDs with different wavelengths and photosensitive elements attached with different filters, or is composed of a xenon lamp and a rotatable filter with a specific wavelength.

3. The system according to claim 1, characterized in that, The broad-spectrum light source is a xenon lamp or an LED array.

4. The system according to claim 1, characterized in that, The spectral directions of the pair of linear gradient filters F1 and F2 are perpendicular.

5. The system according to claim 1, characterized in that, The linear gradient filter is made by an ion-assisted method or an ion beam sputtering method process.

6. The system according to claim 1, characterized in that, The imaging device is a CCD or a CMOS camera.

7. The system according to claim 1, characterized in that, The layout of the collimator and the lens in the optical path is optimized to reduce stray light.

8. The system according to claim 1, characterized in that, This system also includes an efficient spectral data processing algorithm for realizing spectral extraction and analysis.

9. The system according to claim 1, characterized in that, The horizontal direction x corresponds to the spectral distribution of the incident light, and the vertical direction y corresponds to the spectral distribution of the fluorescence or transmitted light emitted by the sample; the upper half matrix (y>x) reflects the fluorescence emission characteristics of the sample at different excitation wavelengths, and the lower half matrix (y<x) reflects the non-fluorescent process.

10. The system according to claim 1, characterized in that, When the included angle between the spectral directions of the linear gradient filters F1 and F2 is less than 90°, the resolution of the emission spectrum is improved; when the included angle is greater than 90°, the resolution of the reception spectrum is improved. The change in the light intensity in the diagonal direction (x = y) reflects the absorption characteristics of the sample to light with different wavelengths.

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