A full spectrum fluorescence lifetime rapid measurement device

Abstract

This invention provides a rapid full-spectrum fluorescence lifetime measurement device, aiming to solve the problems of slow measurement speed and difficulty in simultaneously obtaining fluorescence information at multiple wavelengths due to count rate limitations in time-correlated single-photon counting-based fluorescence lifetime measurement methods. The measurement device includes an optical system and a detection circuit system. The optical system is used to split and shape fluorescence signals of different wavelengths, separating wavelengths in one dimension and uniformly distributing light intensity in another. The detection circuit system is used to receive the shaped two-dimensionally distributed fluorescence signal, perform photon counting, obtain the time difference between the excitation light signal and the fluorescence signal, accumulate the number of fluorescence photons in each pixel unit, obtain a fluorescence lifetime decay histogram at different wavelengths, and finally obtain full-spectrum fluorescence lifetime information. This invention utilizes a time-domain-based fluorescence lifetime measurement method, combining fluorescence spot splitting and shaping with a multi-channel detector and signal processing system, to achieve rapid measurement of the full-spectrum fluorescence lifetime of a sample.

Description

Technical Field

[0001] This invention relates to the field of time-resolved fluorescence lifetime measurement technology, and in particular to a rapid measurement device for full-spectrum fluorescence lifetime. Background Technology

[0002] Under the illumination of an excitation source, molecules in a substance absorb energy and transition to an excited state, then return to the ground state via radiative transition, releasing photons and emitting fluorescence. The fluorescence lifetime of a substance is related to the polarity and viscosity of its microenvironment, and is not affected by fluorophore concentration, sample thickness, photobleaching, or excitation light intensity. Therefore, measuring the fluorescence lifetime of a sample can provide parameters of the molecular microenvironment, such as pH and ion concentration, enabling more precise and in-depth functional measurements of biological samples. Subtle changes in emission spectra and fluorescence lifetimes can reveal intermolecular interactions and the influence of environmental parameters, such as fluorescence resonance energy transfer and temperature.

[0003] Among existing fluorescence lifetime measurement techniques, time-correlated single-photon counting (TRCS) offers advantages such as high measurement accuracy and suitability for measuring the fluorescence lifetime of multi-component samples and those with weak fluorescence intensity. The measurement requires synchronized pulsed electrical signals and pulsed lasers. After the measurement begins, the synchronized pulsed electrical signal triggers a timer, and simultaneously, the pulsed laser excites the sample to emit fluorescence. The timer stops when the first fluorescent photon signal reaches the detector. The timer records the time interval between the arrival of the synchronized pulsed electrical signal and the fluorescent photon signal at the detector, and includes this time interval in the corresponding time channel within the pulse period. Therefore, under high-repetition-rate pulsed laser excitation, after a certain accumulation time, a histogram is obtained with time on the x-axis and photon count on the y-axis. After smoothing, a fluorescence decay curve is obtained, as shown below. Figure 1 As shown.

[0004] Within an excitation cycle, a typical single-channel time-correlated single-photon counting method records at most one photon. If multiple photons occur within a single excitation cycle, the measured fluorescence lifetime will shift towards a shorter lifetime compared to the true fluorescence lifetime, a phenomenon known as the "photon accumulation effect." This photon accumulation effect limits the count rate of time-correlated single-photon counting. Typically, the ratio of the fluorescence photon count rate to the excitation pulse repetition frequency needs to be below 1%–5%, otherwise, fluorescence lifetime measurement will be distorted. Therefore, to ensure a sufficient number of photons, a sufficiently long measurement time is required to improve the accuracy of the results. On the other hand, in single-channel time-correlated single-photon counting, a single measurement often only detects a single wavelength of a sample, making it impossible to simultaneously obtain fluorescence lifetime information at other detection wavelengths. To obtain lifetime information for the same sample at multiple detection wavelengths, multiple measurements of the sample are required. This significantly limits the application of fluorescence lifetime in studies such as fluorescence resonance energy transfer and interactions between multiple fluorescent substances.

[0005] Therefore, there is a need for a full-spectrum fluorescence lifetime measurement device that can measure fluorescence information at multiple wavelengths simultaneously with high speed. Summary of the Invention

[0006] To address the shortcomings of existing technologies, this invention provides a rapid measurement device for full-spectrum fluorescence lifetime.

[0007] The present invention provides a rapid measurement device for full-spectrum fluorescence lifetime, comprising: an optical system and a detection circuit system.

[0008] The optical system includes, in sequence along the optical path: a collimation component for collimating fluorescence into an approximately parallel beam; a beam splitting component for separating fluorescence of different wavelengths in one dimension, i.e., the X direction; and an optical shaping component for shaping a fluorescence spot with a Gaussian intensity distribution into a spot with a uniform intensity distribution, i.e., a rectangular fluorescence spot, in another dimension, i.e., the Y direction.

[0009] The detection circuit system includes a multi-channel detector and a signal processing system, and performs the following steps:

[0010] The rectangular fluorescent spot, processed by the optical system, is received. This rectangular fluorescent spot exhibits wavelength separation in one dimension (X-direction) and uniform intensity distribution in another dimension (Y-direction). By converting the optical signal into an electrical signal, photon counting is performed on each pixel unit in both the X and Y directions of the multi-channel detector.

[0011] The time difference between the arrival of the electrical pulse synchronization signal and the fluorescence signal for each pixel unit is obtained.

[0012] Based on the photon count of each pixel unit and the time difference between the arrival of the electrical pulse synchronization signal and the fluorescence signal for that pixel unit, a cumulative histogram of the photon count of each pixel unit over time is obtained.

[0013] Each pixel unit in the Y direction of the multi-channel detector receives fluorescence of the same wavelength. By processing the photon count accumulation histogram data in the Y direction in parallel, the fluorescence lifetime decay histogram at the same wavelength is quickly obtained. Different pixel units in the X direction of the multi-channel detector correspond to different fluorescence wavelengths, thereby further quickly obtaining the full-spectrum fluorescence lifetime decay histogram, i.e., full-spectrum fluorescence lifetime information.

[0014] Preferably, in the optical system, the beam-splitting component is a dispersive element, which is a grating or a prism.

[0015] Preferably, in the optical path of the optical system, the optical shaping component is located after the beam splitter, or the optical shaping component is located in front of the beam splitter.

[0016] Preferably, when the optical shaping component is in front of the beam splitter, the optical shaping component is a Powell prism.

[0017] Preferably, when the optical shaping component is located after the beam splitter, the optical shaping component is a plano-concave cylindrical mirror and a Powell prism, or a plano-convex cylindrical mirror and a Powell prism.

[0018] Preferably, the multi-channel detector is a planar array multi-pixel single-photon detector or a two-dimensional combination of multiple single-pixel single-photon detectors.

[0019] Preferably, the multi-channel detector is configured to receive the rectangular light spot and realize photon counting on each pixel unit, and the signal processing system is configured to obtain the time difference between the arrival of the electrical pulse synchronization signal and the fluorescence signal of each pixel unit, and to obtain the cumulative histogram of the photon count of each pixel unit over time; or the multi-channel detector is configured to realize photon counting on each pixel unit and obtain the time difference between the arrival of the electrical pulse synchronization signal and the fluorescence signal of each pixel unit, and the signal processing system is configured to obtain the cumulative histogram of the photon count of each pixel unit over time.

[0020] Preferably, the measuring device can measure fluorescence at the same wavelength without the spectral splitting component. Each pixel unit in the multiple pixel units in the Y direction of the multi-channel detector receives fluorescence at the same wavelength. By processing the photon count accumulation histogram data in the Y direction in parallel, the fluorescence lifetime decay histogram at the same wavelength can be obtained quickly.

[0021] The present invention has the following beneficial effects:

[0022] 1. Compared with the traditional single-channel time-correlated single-photon counting fluorescence lifetime measurement method, this invention overcomes the limitation of low photon count rate in the single-channel time-correlated single-photon counting measurement method by performing fluorescence spectral dispersion and shaping processing and combining it with a multi-channel detector and corresponding circuits. Furthermore, it can simultaneously detect fluorescence at multiple wavelengths, significantly improving the measurement speed of full-spectrum fluorescence lifetime.

[0023] 2. In this invention, the reshaping of the fluorescent spot makes the light intensity distribution of the light signal received by the multi-channel detector uniform in one dimension (Y direction). This effectively avoids the state of saturation caused by the light intensity being strong in the middle and weak at both ends, resulting in the photon count rate being too low at both ends and the photon count rate being too high in the middle channel. This allows all pixel units of the multi-channel detector to be fully utilized in this direction. Attached Figure Description

[0024] Figure 1 This is a schematic diagram illustrating the principle of measuring fluorescence lifetime using time-correlated single-photon counting.

[0025] Figure 2 This is a schematic diagram of the structure of a rapid measurement device for full-spectrum fluorescence lifetime according to Embodiment 1 of the present invention;

[0026] Figure 3 This is a schematic diagram of the structure of a rapid measurement device for full-spectrum fluorescence lifetime according to Embodiment 2 of the present invention;

[0027] Figure 4 is a simulation result of fluorescence spot information with wavelengths of 500-700 nm on a multi-channel detector in an embodiment of the present invention. (a) is a wavelength distribution diagram, and (b) is an intensity distribution diagram. The wavelength interval between adjacent fluorescence spots is 10 nm.

[0028] Figure 5 This is a schematic diagram of the structure of a rapid full-spectrum fluorescence lifetime measurement system according to Embodiment 3 of the present invention;

[0029] Figure 6 This is a schematic diagram of the structure of a full-spectrum fluorescence lifetime rapid imaging system according to Embodiment 4 of the present invention.

[0030] In the figure, 1 is the collimating mirror, 2 is the first reflecting mirror, 3 is the beam splitting assembly, 4 is the focusing assembly, 5 is the plano-concave cylindrical mirror, 6 is the Powell prism, 7 is the multi-channel detector, 8 is the signal processing system, 9 is the pulsed laser, 10 is the dichroic mirror, 11 is the second reflecting mirror, 12 is the first focusing lens, 13 is the sample cell, 14 is the second focusing lens, 15 is the X-axis motor and its reflecting mirror, 16 is the Y-axis motor and its reflecting mirror, 17 is the scanning lens, 18 is the sleeve lens, 19 is the pinhole, and 20 is the host computer. Detailed Implementation

[0031] The embodiments of the present invention will be described below with reference to the accompanying drawings. It should be noted that the embodiments described in this specification are not exhaustive and do not represent the only embodiments of the present invention. The corresponding embodiments below are only for clearly illustrating the inventive content of this patent and are not intended to limit its implementation. For those skilled in the art, different variations and modifications can be made based on these embodiments. Any variations or modifications that fall within the technical concept and inventive content of the present invention and are obvious are also within the protection scope of the present invention.

[0032] Figure 2 This is a schematic diagram of a rapid full-spectrum fluorescence lifetime measurement device according to Embodiment 1 of the present invention. The rapid full-spectrum fluorescence lifetime measurement device in this embodiment includes an optical system and a detection circuit system.

[0033] The optical system comprises, in sequence along the optical path, a collimation component, a beam splitter, and an optical shaping component. The collimation component collimates the fluorescence into an approximately parallel beam. Here, the fluorescence is generated by the light source irradiating the sample. Preferably, the collimation component is a collimating lens. The beam splitter separates fluorescence of different wavelengths in one dimension, i.e., the X-direction. Preferably, the beam splitter is a dispersive element, preferably a grating or prism. The optical shaping component shapes the fluorescence spot with a Gaussian intensity distribution into a spot with a uniform intensity distribution in another dimension, i.e., the Y-direction. In this embodiment, the optical shaping component is located after the beam splitter. Preferably, when the optical shaping component is located after the beam splitter, it is a plano-concave cylindrical mirror and a Powell prism, or a plano-convex cylindrical mirror and a Powell prism.

[0034] The detection circuit system includes a multi-channel detector and a signal processing system. In this embodiment, the multi-channel detector receives the rectangular light spot and performs photon counting on each pixel unit. The signal processing system obtains the time difference of each pixel unit, accumulates the fluorescence signal, and thus obtains the fluorescence lifetime decay results at different wavelengths, achieving rapid measurement of the full-spectrum fluorescence lifetime. In another embodiment, the timing function for obtaining the time difference of each pixel unit can also be implemented by the multi-channel detector.

[0035] The following is combined Figure 2 Provide a detailed description of the optical system. For example... Figure 2 As shown, fluorescence enters the optical system of the full-spectrum fluorescence lifetime rapid measurement device. After being reflected by collimating lens 1, it becomes an approximately parallel beam and reaches the beam splitter 3 through the first reflecting mirror 2. After being split by the beam splitter 3, fluorescence of different wavelengths is separated in the X direction and reflected to the focusing component 4. After separation, the fluorescence of each wavelength converges in the X and Y directions through the focusing component 4. Subsequently, it passes through the plano-concave cylindrical mirror 5, which diverges only in the Y direction, and becomes a beam collimated in the Y direction and converged in the X direction, completing the fluorescence beam splitting process. After collimation in the Y direction, the fluorescence of each wavelength enters the Powell prism 6, which shapes the circular light spots of each wavelength into rectangular light spots with uniform intensity distribution in the Y direction, completing the fluorescence spot shaping process. The plano-concave cylindrical mirror 5 and the Powell prism 6 are located in front of the focal position of the focusing component 4, wherein the focal position of the plano-concave cylindrical mirror 5 is the same as the focal position of the focusing component 4.

[0036] After being processed by the optical system, the fluorescence reaches the detection circuit system. The detection circuit system includes a multi-channel detector 7 and a signal processing system 8. The detection circuit system is described in detail below.

[0037] A rectangular fluorescent spot with wavelengths separated along the X direction and uniform light intensity distribution along the Y direction is formed on the multi-channel detector 7, as shown in Figures 4(a) and 4(b). The multi-channel detector 7 is located at the focal point of the focusing component 4 in the optical system. The multi-channel detector 7 receives the rectangular fluorescent spot, processes the optical signal into a digital signal, and performs photon counting on each pixel unit. In each preferred embodiment, the multi-channel detector 7 is a planar array multi-pixel single-photon detector or a two-dimensional combination of multiple single-pixel single-photon detectors. That is, the multi-channel detector 7 has multiple pixel units in both the X and Y directions, and each pixel unit can be represented as I. (x,y) In this way, the multi-channel detector 7 can simultaneously detect fluorescence of multiple wavelengths in the X direction, and it can also utilize multiple pixel units to simultaneously count photons of the same wavelength of fluorescence in the Y direction. For example, at position x1, I... (x1，y1) ~I (x1，yn) Multiple pixel units simultaneously count photons of fluorescence at the same wavelength.

[0038] The signal processing system 8 obtains the time difference between the arrival of the electrical pulse synchronization signal and the fluorescence signal for each pixel unit. Furthermore, based on the photon count for each pixel unit and the corresponding time difference between the arrival of the electrical pulse synchronization signal and the fluorescence signal for each pixel unit, it obtains a cumulative histogram of the photon count over time for each pixel unit.

[0039] In this multi-channel detector, each pixel unit in the Y-direction receives fluorescence of the same wavelength. Each pixel unit can independently perform photon counting and timing functions. The output signals of all pixel units in this direction are transmitted in parallel to the data acquisition unit for parallel processing. In this way, the signal processing task is distributed among multiple pixel units in the Y-direction, reducing the photon counting load of each pixel unit. This avoids the photon accumulation effect in Time-Correlated Single Photon Counting (TCSPC) and enables rapid readout of pixel data, thereby quickly obtaining the fluorescence lifetime decay histogram at that wavelength. In addition, through the fluorescence spot shaping process, the light intensity distribution of the light signal received by the multi-channel detector is made uniform in one dimension (Y-direction). This effectively avoids the state of excessively low photon count rates at both ends and excessively high photon count rates in the middle channel caused by strong light intensity in the middle and weak light intensity at both ends. This ensures that all pixel units in the Y-direction of the multi-channel detector are fully utilized.

[0040] Furthermore, in the X direction of the multi-channel detector, by utilizing the characteristic that different pixels in the X direction of the multi-channel detector 7 correspond to different fluorescence wavelengths, the full-spectrum fluorescence lifetime decay histogram, i.e., full-spectrum fluorescence lifetime information, can be obtained more quickly.

[0041] In the optical system of the full-spectrum fluorescence lifetime rapid measurement device described in this embodiment, the plano-concave cylindrical mirror 5, which diverges the beam only in the Y direction, can also be a plano-convex cylindrical mirror that converges the beam only in the Y direction. The plano-convex cylindrical mirror is located behind the focal position of the focusing component 4, and its focal position is the same as that of the focusing component 4. A Powell prism 6 for shaping the Y direction and a multi-channel detector 7 are placed sequentially behind the plano-convex cylindrical mirror. A rectangular fluorescence spot with wavelengths separated in the X direction and uniform light intensity distribution in the Y direction is formed on the multi-channel detector 7.

[0042] Figure 3 This is a schematic diagram of a rapid full-spectrum fluorescence lifetime measurement device according to Embodiment 2 of the present invention. The rapid full-spectrum fluorescence lifetime measurement device in this embodiment includes an optical system and a detection circuit system.

[0043] The optical system comprises, in sequence along the optical path, a collimating component, a beam splitting component, and an optical shaping component. The collimating component collimates the fluorescence into an approximately parallel beam. Preferably, the collimating component is a collimating lens. The beam splitting component separates fluorescence of different wavelengths in one dimension, i.e., the X-direction. Preferably, the beam splitting component is a dispersive element, preferably a grating or a prism. The optical shaping component shapes the fluorescence spot with a Gaussian intensity distribution into a spot with a uniform intensity distribution in another dimension, i.e., the Y-direction. In this embodiment, the optical shaping component is located before the beam splitting component. Preferably, when the optical shaping component is located before the beam splitting component, the optical shaping component is a Powell prism.

[0044] The following is combined Figure 3 Provide a detailed description of the optical system. For example... Figure 3 As shown, the fluorescence enters the optical system of the full-spectrum fluorescence lifetime rapid measurement device. After being reflected by the collimating lens 1, it becomes an approximately parallel beam and enters the optical path of the first reflecting mirror 2. The Powell prism 6, which shapes the Y-direction, reshapes the circular spot into a rectangular spot with uniform intensity distribution in the Y-direction, completing the fluorescence spot shaping process. The shaped fluorescence enters the beam splitter 3. After being split by the beam splitter 3, fluorescence of different wavelengths is separated in the X-direction and reflected to the focusing component 4. After separation, the fluorescence of each wavelength converges in the X and Y directions through the focusing component 4, completing the fluorescence beam splitting process.

[0045] After being processed by the optical system, the fluorescence enters the detection circuit system. The detection circuit system includes a multi-channel detector 7 and a signal processing system 8. The structure of the detection circuit system in this embodiment is the same as that in Embodiment 1, and will not be described again here.

[0046] Figure 5 This is a schematic diagram of a rapid full-spectrum fluorescence lifetime measurement system according to Embodiment 3 of the present invention. The measurement system includes... Figure 2The full-spectrum fluorescence lifetime rapid measurement device shown can realize the rapid measurement of the full-spectrum fluorescence lifetime of a sample.

[0047] like Figure 5 As shown, the excitation light emitted by the pulsed laser 9 enters the fluorescence excitation and collection components 10-19, is reflected when it reaches the dichroic mirror 10, and enters the first focusing lens 12 through the second reflecting mirror 11. The first focusing lens 12 focuses the excitation light onto the sample surface in the sample cell 13, completing the sample excitation. The fluorescence emitted by the sample returns along the original path, is transmitted through the dichroic mirror 10, and enters the full-spectrum fluorescence lifetime rapid measurement device 1-8 through the second focusing lens 14. The optical system performs fluorescence spectral dispersion and spot shaping, and the detection circuit system performs rapid detection and transmission of fluorescence information. The signal processing system 8 simultaneously monitors the transmitted digital signal and the pulse synchronization signal of the pulsed laser 9, and transmits the information to the host computer 20. The host computer 20 performs statistical analysis and further calculations on the sample fluorescence information to obtain the full-spectrum fluorescence lifetime information of the sample.

[0048] Preferably, in the fluorescence excitation and collection components 10-19, the sample cell 13 can be a two-dimensional displacement stage, wherein the two-dimensional displacement stage can move in two dimensions in a direction parallel to the horizontal plane to realize the movement of the sample. For example, the two-dimensional displacement stage controlled by the host computer 20 can be used to realize the rapid measurement of the full spectrum fluorescence lifetime of different samples in a multi-well plate.

[0049] Figure 6 This is a schematic diagram of the structure of a full-spectrum fluorescence lifetime rapid imaging system according to Embodiment 4 of the present invention. The imaging system includes... Figure 3 The full-spectrum fluorescence lifetime rapid measurement device shown can realize rapid imaging of the full-spectrum fluorescence lifetime of samples.

[0050] like Figure 6 As shown, the excitation light emitted by the pulsed laser 9 enters the fluorescence excitation and collection components 10-19, is reflected when it reaches the dichroic mirror 10, and passes through the second reflecting mirror 11 into the two-dimensional galvanometer system composed of the X-axis motor and its reflecting mirror 15 and the Y-axis motor and its reflecting mirror 16. It then passes sequentially through the scanning lens 17 and the sleeve lens 18, and is focused by the first focusing lens 12 onto the sample surface in the sample cell 13, completing the sample excitation. The fluorescence emitted by the sample returns along the original path, is transmitted through the dichroic mirror 10, and enters the full-spectrum fluorescence lifetime rapid measurement device 1-8 via the second focusing lens 14 and the pinhole 19. The optical system performs fluorescence dispersion and spot shaping, and the detection circuit system performs rapid detection and transmission of fluorescence information. The signal processing system 8 simultaneously monitors the transmitted digital signal and the pulse synchronization signal of the pulsed laser 9, and transmits the information to the host computer 20. The host computer 20 performs statistical analysis and further calculations on the sample fluorescence information to obtain the full-spectrum fluorescence lifetime information of the sample.

[0051] Preferably, in the fluorescence excitation and collection components 10-19, the X-axis motor and its reflector 15, the Y-axis motor and its reflector 16, the scanning lens 17, the sleeve lens 18, and the pinhole 19 are laser confocal scanning microscopy imaging components, which can be controlled by the host computer 20 to achieve rapid imaging of the full spectrum fluorescence lifetime of the sample.

[0052] Obviously, those skilled in the art should recognize that the above embodiments are only used to illustrate the present invention and are not intended to limit the present invention. Any changes or modifications to the above embodiments that are within the essential spirit of the present invention will fall within the scope of the claims of the present invention.

Claims

1. A rapid measurement device for full-spectrum fluorescence lifetime, characterized in that, Including optical systems and detection circuit systems, The optical system comprises, along the optical path, the following components in sequence: Collimation components are used to collimate fluorescence into an approximately parallel beam; A spectrometer is used to separate fluorescence of different wavelengths in one dimension, namely the X direction. An optical shaping component is used to shape a fluorescent spot with a Gaussian distribution of light intensity into a rectangular fluorescent spot with a uniform distribution of light intensity in another dimension, namely the Y direction. The detection circuit system includes a multi-channel detector and a signal processing system, and performs the following steps: The rectangular fluorescent spot, processed by the optical system, is received. This rectangular fluorescent spot exhibits wavelength separation in one dimension (X-direction) and uniform intensity distribution in another dimension (Y-direction). By converting the optical signal into an electrical signal, photon counting is performed on each pixel unit in both the X and Y directions of the multi-channel detector. The time difference between the arrival of the electrical pulse synchronization signal and the fluorescence signal for each pixel unit is obtained. Based on the photon count of each pixel unit and the time difference between the arrival of the electrical pulse synchronization signal and the fluorescence signal for that pixel unit, a cumulative histogram of the photon count of each pixel unit over time is obtained. Each pixel unit in the Y direction of the multi-channel detector receives fluorescence of the same wavelength. By processing the photon count accumulation histogram data in the Y direction in parallel, the fluorescence lifetime decay histogram at the same wavelength is quickly obtained. Different pixel units in the X direction of the multi-channel detector correspond to different fluorescence wavelengths, thereby further quickly obtaining the full-spectrum fluorescence lifetime decay histogram, i.e., full-spectrum fluorescence lifetime information.

2. The apparatus according to claim 1, wherein In the optical system, the beam-splitting component is a dispersive element, which is a grating or a prism.

3. The rapid measurement device for full-spectrum fluorescence lifetime according to claim 1, characterized in that, In the optical path of the optical system, the optical shaping component is located after the beam splitter, or the optical shaping component is located in front of the beam splitter.

4. The apparatus according to claim 3, wherein When the optical shaping component is in front of the beam splitter, the optical shaping component is a Powell prism.

5. The apparatus according to claim 3, wherein When the optical shaping component is located after the beam splitter, the optical shaping component is a plano-concave cylindrical mirror and a Powell prism, or a plano-convex cylindrical mirror and a Powell prism.

6. The apparatus according to claim 1, wherein The multi-channel detector is a planar array multi-pixel single-photon detector or a two-dimensional combination of multiple single-pixel single-photon detectors.

7. The apparatus of claim 1, wherein the apparatus is a full spectrum fluorescence lifetime rapid measurement device. The multi-channel detector is configured to receive the rectangular light spot and count photons on each pixel unit. The signal processing system is configured to obtain the time difference between the arrival of the electrical pulse synchronization signal and the fluorescence signal for each pixel unit, and to obtain a cumulative histogram of the photon count of each pixel unit over time. Alternatively, the multi-channel detector is configured to count photons on each pixel unit and obtain the time difference between the arrival of the electrical pulse synchronization signal and the fluorescence signal for each pixel unit, and the signal processing system is configured to obtain a cumulative histogram of the photon count of each pixel unit over time.

8. The apparatus of claim 1, wherein the apparatus is a full spectrum fluorescence lifetime fast measurement device. The measuring device can measure fluorescence at the same wavelength without the spectral splitting component. Each pixel unit in the multiple pixel units in the Y direction of the multi-channel detector receives fluorescence at the same wavelength. By processing the photon count accumulation histogram data in the Y direction in parallel, the fluorescence lifetime decay histogram at the same wavelength can be obtained quickly.

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