CN116148232A - Full spectrum fluorescence life-span rapid measurement device - Google Patents
Full spectrum fluorescence life-span rapid measurement device Download PDFInfo
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- CN116148232A CN116148232A CN202310155494.XA CN202310155494A CN116148232A CN 116148232 A CN116148232 A CN 116148232A CN 202310155494 A CN202310155494 A CN 202310155494A CN 116148232 A CN116148232 A CN 116148232A
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6408—Fluorescence; Phosphorescence with measurement of decay time, time resolved fluorescence
Abstract
The invention provides a full-spectrum fluorescence lifetime rapid measuring device, which aims to solve the problems that the measurement speed is low and fluorescence information under a plurality of wavelengths is difficult to obtain simultaneously due to the limitation of the counting rate in a fluorescence lifetime measuring method based on time-dependent single photon counting. The measuring device includes an optical system and detection circuitry. The optical system is used for carrying out light splitting and shaping on fluorescent signals with different wavelengths, so that the fluorescent signals are separated in wavelength in one dimension and the light intensity in the other dimension is uniformly distributed. The detection circuit system is used for receiving the shaped two-dimensional distributed fluorescent signals, carrying out photon counting to obtain the time difference between the excitation light signals and the fluorescent signals, realizing the accumulation of the fluorescent photon number of each pixel unit, obtaining the fluorescence lifetime attenuation histogram under different wavelengths, and finally obtaining full-spectrum fluorescence lifetime information. The invention utilizes a fluorescence lifetime measurement means based on a time domain, and can realize the rapid measurement of the full-spectrum fluorescence lifetime of the sample by combining the fluorescence spot light splitting and shaping with the multichannel detector and the signal processing system.
Description
Technical Field
The invention relates to the technical field of time-resolved fluorescence lifetime measurement, in particular to a full-spectrum fluorescence lifetime rapid measuring device.
Background
Under the irradiation of an excitation light source, molecules in the substance absorb energy and then transition to an excited state, and then return to a ground state in a radiation transition mode, and fluorescence is emitted along with the release of photons. The fluorescence lifetime of the substance is related to the polarity, viscosity and the like of the microenvironment in which the substance is positioned, and is not influenced by the concentration of fluorophores, the thickness of the sample, the photo bleaching, the intensity of excitation light and the like, so that the parameters of the microenvironment in which the molecules are positioned, such as the pH value, the ion concentration and the like, can be obtained by measuring the fluorescence lifetime of the sample, and the functional accurate measurement of biological samples and the like can be performed more deeply. The effects of intermolecular interactions and environmental parameters, such as fluorescence resonance energy transfer, temperature, etc., can be inferred from subtle changes in emission spectra and fluorescence lifetime.
In the existing fluorescence lifetime measurement technology, time-dependent single photon counting has the advantages of high measurement precision and suitability for measuring samples with weak fluorescence lifetime and fluorescence intensity. Synchronous pulsed electrical signals and pulsed lasers are required for measurement. The synchronous pulse electric signal triggers the timer to start timing after the measurement is started, meanwhile, the pulse laser excites the sample to emit fluorescence, and the timing is stopped after the first fluorescence photon signal reaches the detector. The timer will record the time interval between arrival of the synchronous pulse electrical signal and the fluorescent photon signal at the detector and count it in the corresponding time channel within the pulse period. Therefore, after a certain time of accumulation under the excitation of the pulse laser with high repetition frequency, a histogram with time on the abscissa and photon number on the ordinate is obtained, and a fluorescence attenuation curve is obtained after the smoothing treatment, as shown in fig. 1.
A typical single channel time-dependent single photon count records at most one photon during one excitation period. If multiple photons occur in one excitation period, the measured fluorescence lifetime is shifted in the short lifetime direction compared to the real fluorescence lifetime, i.e. a "photon stacking effect" occurs. This photon packing effect limits the count rate of time-dependent single photon counting methods. Typically the ratio of the fluorescent photon count rate to the excitation pulse repetition frequency needs to be below 1% to 5%, otherwise it can lead to distortion of the fluorescent lifetime measurement. Therefore, to ensure a sufficient number of photons, a long enough measurement time is required to improve the accuracy of the result. On the other hand, in the single-channel time-dependent single photon counting method, only a single wavelength of a certain sample can be detected by one measurement, and fluorescence lifetime information under other detection wavelengths can not be obtained at the same time. To obtain lifetime information for multiple probe wavelengths for the same sample, multiple measurements of the sample are required. This greatly limits the use of fluorescence lifetime in studies of fluorescence resonance energy transfer, interactions of various fluorescent substances, etc.
Therefore, there is a need for a full spectrum fluorescence lifetime measurement device that is fast in measurement speed and capable of detecting fluorescence information at a plurality of wavelengths simultaneously.
Disclosure of Invention
The invention provides a full-spectrum fluorescence lifetime rapid measuring device for solving the defects in the prior art.
The invention relates to a full spectrum fluorescence lifetime rapid measuring device, which comprises: an optical system and a detection circuitry system,
the optical system sequentially comprises the following components along an optical path: a collimation assembly for collimating the fluorescence into an approximately parallel beam; the light splitting component is used for splitting fluorescence with different wavelengths in one-dimensional direction, namely X direction; the optical shaping component is used for shaping the fluorescent light spots with Gaussian light intensity distribution into light spots with uniform light intensity distribution, namely rectangular fluorescent light spots, in the other dimension direction, namely the Y direction;
the detection circuitry includes a multi-channel detector and a signal processing system and performs the steps of:
receiving the rectangular fluorescent light spots processed by the optical system, wherein the rectangular fluorescent light spots are separated in wavelength in one dimension, namely the X direction, have uniform light intensity distribution in the other dimension, namely the Y direction, and photon counting is carried out on each of a plurality of pixel units in the X direction and the Y direction of the multichannel detector by converting optical signals into electric signals,
obtaining the time difference between the arrival of the electric pulse synchronous signal and the fluorescent signal of each pixel unit,
obtaining a cumulative histogram of photon numbers of each pixel unit along with time channels according to photon counts of each pixel unit and time differences between arrival of corresponding electric pulse synchronous signals and fluorescence signals of each pixel unit,
each pixel unit in a plurality of pixel units in the Y direction of the multichannel detector receives fluorescence of the same wavelength, and the fluorescence lifetime attenuation histogram under the same wavelength is rapidly obtained by processing photon number cumulative histogram data in the Y direction in parallel; different pixel units of the multichannel detector in the X direction correspond to different fluorescence wavelengths, so that a full-spectrum fluorescence lifetime attenuation histogram, namely full-spectrum fluorescence lifetime information, is further and rapidly obtained.
Preferably, in the optical system, the light splitting component is a dispersive element, and the dispersive element is a grating or a prism.
Preferably, in the optical path of the optical system, the optical shaping component is behind the beam splitting component, or the optical shaping component is in front of the beam splitting component.
Preferably, the optical shaping component is a powell lens when the optical shaping component is in front of the beam splitting component.
Preferably, the optical shaping component is a plano-concave cylindrical mirror and a powell lens or a plano-convex cylindrical mirror and a powell lens when the optical shaping component is behind the beam splitting component.
Preferably, the multi-channel detector is a planar array type multi-pixel single photon detector or a two-dimensional combination of a plurality of single pixel single photon detectors.
Preferably, the multi-channel detector is configured to receive the rectangular light spot, to effect photon counting on each pixel cell, and the signal processing system is configured to obtain a time difference between arrival of the electrical pulse synchronization signal and the fluorescence signal for each pixel cell, and to obtain a cumulative histogram of photon count for each pixel cell over time channel; or the multi-channel detector is configured to effect photon counting per pixel cell and to obtain a time difference between arrival of the electrical pulse synchronization signal and the fluorescence signal per pixel cell, the signal processing system being configured to obtain a cumulative histogram of photon counts per pixel cell over time channels.
Preferably, the measuring device may measure fluorescence of the same wavelength without the beam splitter, each of the plurality of pixel units in the Y direction of the multi-channel detector receives fluorescence of the same wavelength, and the fluorescence lifetime attenuation histogram in the same wavelength is rapidly obtained by processing the photon number cumulative histogram data in the Y direction in parallel.
The invention has the following beneficial effects:
1. compared with the traditional single-channel time-related single photon counting fluorescence lifetime measurement method, the method breaks through the limitation of low photon counting rate in the single-channel time-related single photon counting measurement method by combining the multi-channel detector and the corresponding circuit through the light splitting and shaping treatment of fluorescence, can detect fluorescence with multiple wavelengths at the same time, and greatly improves the measurement speed of full-spectrum fluorescence lifetime.
2. According to the invention, through shaping treatment of fluorescent light spots, the light intensity distribution of the light signals received by the multichannel detector is uniform in one dimension (in the Y direction), so that the states of excessively low photon count rate at two ends and excessively high and saturated photon count rate in the middle channel of the multichannel detector caused by strong light intensity middle and weak light intensity at two sides are effectively avoided, and all pixel units in the direction of the multichannel detector are fully utilized.
Drawings
FIG. 1 is a schematic diagram of a time-dependent single photon counting method for measuring fluorescence lifetime;
FIG. 2 is a schematic structural diagram of a full spectrum fluorescence lifetime rapid measurement device according to embodiment 1 of the present invention;
FIG. 3 is a schematic structural diagram of a full spectrum fluorescence lifetime rapid measurement device in accordance with embodiment 2 of the present invention;
FIG. 4 is a graph of simulation results of information of fluorescent light spots with wavelengths of 500-700 nm on a multichannel detector according to an embodiment of the present invention, where (a) is a wavelength distribution diagram, (b) is an intensity distribution diagram, and the wavelength interval between adjacent fluorescent light spots is 10nm;
FIG. 5 is a schematic structural diagram of a full spectrum fluorescence lifetime rapid measurement system according to embodiment 3 of the present invention;
fig. 6 is a schematic structural diagram of a full spectrum fluorescence lifetime rapid imaging system in embodiment 4 of the present invention.
In the figure, 1 is a collimating mirror, 2 is a first reflecting mirror, 3 is a light splitting component, 4 is a focusing component, 5 is a plano-concave cylindrical mirror, 6 is a Bowil prism, 7 is a multichannel detector, 8 is a signal processing system, 9 is pulse laser, 10 is a dichroic mirror, 11 is a second reflecting mirror, 12 is a first focusing lens, 13 is a sample cell, 14 is a second focusing lens, 15 is an X-axis motor and reflecting mirror thereof, 16 is a Y-axis motor and reflecting mirror thereof, 17 is a scanning lens, 18 is a sleeve lens, 19 is a small hole, and 20 is an upper computer.
Detailed Description
Embodiments of the present invention will be described below with reference to the drawings. The embodiments described in the present specification are not intended to be exhaustive or to represent the only embodiments of the present invention. The following examples are presented for clarity of illustration of the invention of the present patent and are not intended to limit the embodiments thereof. Various changes and modifications may be made by one of ordinary skill in the art in light of the above description, and it is intended to cover all such changes and modifications as fall within the true spirit and scope of the invention.
Fig. 2 is a schematic structural diagram of a full spectrum fluorescence lifetime rapid measurement device in embodiment 1 of the present invention. The full-spectrum fluorescence lifetime rapid measurement device in the embodiment comprises an optical system and a detection circuit system.
The optical system sequentially comprises the following components along an optical path: a collimation assembly, a beam splitting assembly and an optical shaping assembly. The collimation assembly is used to collimate the fluorescence light into a nearly parallel beam. Here, fluorescence is generated by excitation after the sample is irradiated by a light source. Preferably, the collimating assembly is a collimating mirror. And the light splitting assembly is used for splitting fluorescence with different wavelengths in one-dimensional direction, namely X direction. Preferably, the light splitting component is a dispersive element, preferably a grating or a prism. And the optical shaping component is used for shaping the fluorescent light spots with Gaussian light intensity distribution into light spots with uniform light intensity distribution in the other dimension direction, namely the Y direction. In this embodiment, the optical shaping element is behind the light splitting element. Preferably, the optical shaping component is a plano-concave cylindrical mirror and a powell lens or a plano-convex cylindrical mirror and a powell lens when the optical shaping component is behind the beam splitting component.
The detection circuitry includes a multi-channel detector and a signal processing system. In this embodiment, the multi-channel detector receives the rectangular light spot, so as to implement a photon counting function on each pixel unit, and the signal processing system obtains a time difference of each pixel unit, and performs fluorescence signal accumulation, so as to obtain fluorescence lifetime attenuation results of different wavelengths, and realize rapid measurement of full-spectrum fluorescence lifetime. In further embodiments, the timing function of obtaining the time difference for each pixel cell may also be implemented by a multi-channel detector.
The optical system is described in detail below with reference to fig. 2. As shown in fig. 2, fluorescence enters an optical system of the full-spectrum fluorescence lifetime rapid measuring device, becomes approximately parallel light beams after being reflected by the collimator lens 1, and reaches the beam splitting assembly 3 through the first reflector 2, fluorescence of different wavelengths is split in the X direction after the beam splitting treatment of the beam splitting assembly 3, and is reflected to the focusing assembly 4, and fluorescence of each wavelength after the splitting is converged in the X direction and the Y direction through the focusing assembly 4. The spectroscopic processing of fluorescence is then completed by the plano-concave cylindrical mirror 5 that diverges the light beam in only the Y direction, becoming a light beam collimated in the Y direction and converging in the X direction. And after the fluorescent light of each wavelength is collimated in the Y direction, the fluorescent light enters a Bowilt prism 6 for shaping the Y direction, and the round light spots of each wavelength are shaped into rectangular light spots with uniform light intensity distribution in the Y direction, so that the light spot shaping treatment of the fluorescent light is completed. The plano-concave cylindrical mirror 5 and the powell edge 6 are positioned in front of the focal position of the focusing assembly 4, wherein the focal position of the plano-concave cylindrical mirror 5 is the same as the focal position of the focusing assembly 4.
After being processed by the optical system, the fluorescence reaches the detection circuit system. The detection circuitry comprises a multi-channel detector 7 and a signal processing system 8. The detection circuitry is described in detail below.
Rectangular fluorescent light spots, which are wavelength-separated in the X-direction and uniformly distributed in the Y-direction in light intensity, are formed on the multi-channel detector 7, as shown in fig. 4 (a) and 4 (b), wherein the multi-channel detector 7 is located at the focal position of the focusing assembly 4 in the optical system. The multi-channel detector 7 is configured to receive the rectangular fluorescent light spot, process the optical signal into a digital signal, and perform 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. I.e. the multi-channel detector 7 has a plurality of pixel cells in both the X-direction and the Y-direction, each pixel cell being denoted I (x,y) . In this way, the multichannel detector 7 can detect fluorescence of a plurality of wavelengths at the same time in the X direction, and the multichannel detector 7 can count photons of fluorescence of the same wavelength at the same time by using a plurality of pixel units in the Y direction. For example, at the x1 position, I can be utilized (x1，y1) ～I (x1，yn) Photon counting is performed on fluorescence light of the same wavelength at the same time.
The signal processing system 8 obtains the time difference of arrival of the electrical pulse synchronization signal and the fluorescence signal for each pixel cell. And obtaining an accumulated histogram of photon numbers under each pixel unit along with a time channel according to photon counts of each pixel unit and the corresponding time difference between arrival of the electric pulse synchronous signals and the fluorescence signals of each pixel unit.
Each pixel unit in a plurality of pixel units in the Y direction of the multichannel detector receives fluorescence with the same wavelength, each pixel unit can independently perform functions such as photon counting and timing, output signals of all pixel units in the Y direction are transmitted to a data collector in parallel for parallel processing, thus a signal processing task is distributed to the plurality of pixel units in the Y direction, photon counting load of each pixel unit is reduced, photon accumulation effect in a time-dependent single photon counting Technology (TCSPC) can be avoided, pixel data can be read out quickly, and a fluorescence lifetime attenuation histogram under the wavelength can be obtained quickly. In addition, the shaping treatment of the fluorescent light spots ensures that the light intensity distribution of the light signals received by the multichannel detector is uniform in one dimension (in the Y direction), and the states of excessively low photon count rate at two ends and excessively high and saturated photon count rate in the middle channel of the multichannel detector caused by strong middle and weak two sides of the light intensity are effectively avoided, so that all pixel units of the multichannel detector in the Y direction are fully utilized.
In addition, in the X direction of the multichannel detector, the characteristic that different pixels in the X direction of the multichannel detector 7 correspond to different fluorescence wavelengths is utilized, so that the full-spectrum fluorescence lifetime attenuation histogram, namely full-spectrum fluorescence lifetime information, is further and rapidly obtained.
In the optical system of the full-spectrum fluorescence lifetime rapid measurement device of this embodiment, the plano-concave cylindrical mirror 5 that only diverges light beams in the Y direction may be a plano-convex cylindrical mirror that only converges light beams in the Y direction, where after the plano-convex cylindrical mirror is located at the focal position of the focusing assembly 4, the focal position of the plano-convex cylindrical mirror is identical to the focal position of the focusing assembly 4, and a powell prism 6 and a multi-channel detector 7 that reshape light beams in the Y direction are sequentially placed behind the plano-convex cylindrical mirror, and rectangular fluorescence spots that are separated in wavelength in the X direction and uniformly distributed in light intensity in the Y direction are formed on the multi-channel detector 7.
Fig. 3 is a schematic structural diagram of a full spectrum fluorescence lifetime rapid measurement device in embodiment 2 of the present invention. The full-spectrum fluorescence lifetime rapid measurement device in the embodiment comprises an optical system and a detection circuit system.
The optical system sequentially comprises the following components along an optical path: a collimation assembly, a beam splitting assembly and an optical shaping assembly. The collimation assembly is used to collimate the fluorescence light into a nearly parallel beam. Preferably, the collimating assembly is a collimating mirror. And the light splitting assembly is used for splitting fluorescence with different wavelengths in one-dimensional direction, namely X direction. Preferably, the light splitting component is a dispersive element, preferably a grating or a prism. And the optical shaping component is used for shaping the fluorescent light spots with Gaussian light intensity distribution into light spots with uniform light intensity distribution in the other dimension direction, namely the Y direction. In this embodiment, the optical shaping element is before the light splitting element. Preferably, the optical shaping component is a powell lens when the optical shaping component is in front of the beam splitting component.
The optical system is described in detail below with reference to fig. 3. As shown in fig. 3, fluorescence enters an optical system in the full-spectrum fluorescence lifetime rapid measuring device, becomes approximately parallel light beams after being reflected by the collimating mirror 1, enters an optical path where the first reflecting mirror 2 is located, and a powell lens 6 for shaping a circular light spot into a rectangular light spot with uniform light intensity distribution in the Y direction, so that the light spot shaping treatment of fluorescence is completed. The shaped fluorescence enters the light splitting assembly 3, after the light splitting treatment of the light splitting assembly 3, the fluorescence with different wavelengths is split in the X direction and reflected to the focusing assembly 4, and the fluorescence with each wavelength after the splitting is converged in the X direction and the Y direction through the focusing assembly 4, so that the light splitting treatment of the fluorescence is completed.
After being processed by the optical system, the fluorescence enters the detection circuit system. The detection circuitry comprises 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 of embodiment 1, and will not be described here again.
Fig. 5 is a schematic structural diagram of a full spectrum fluorescence lifetime rapid measurement system in embodiment 3 of the present invention. The measuring system comprises the full-spectrum fluorescence lifetime rapid measuring device shown in fig. 2, and can realize the full-spectrum fluorescence lifetime rapid measurement of a sample.
As shown in fig. 5, the excitation light emitted by the pulse laser 9 enters the fluorescence excitation and collection assembly 10-19, is reflected when reaching the dichroic mirror 10, enters the first focusing lens 12 through the second reflecting mirror 11, the first focusing lens 12 converges the excitation light to the sample surface in the sample cell 13, excitation of the sample is completed, the fluorescence primary path emitted by the sample returns, is transmitted through the dichroic mirror 10, enters the full-spectrum fluorescence lifetime rapid measuring device 1-8 through the second focusing lens 14, performs light splitting and spot shaping treatment of fluorescence through an optical system, and performs rapid detection and transmission of fluorescence information through a detection circuit system. The signal processing system 8 monitors the transmitted digital signal and the pulse synchronous signal of the pulse laser 9 at the same time, and transmits the information to the upper computer 20, and the upper computer 20 completes statistics and further calculation of the fluorescence information of the sample to obtain the full spectrum fluorescence lifetime information of the sample.
Preferably, in the fluorescence excitation and collection assemblies 10-19, the sample cell 13 may be a two-dimensional displacement table, wherein the two-dimensional displacement table may perform two-dimensional movement in a direction parallel to a horizontal plane for realizing movement of the sample, for example, full-spectrum fluorescence lifetime rapid measurement of different samples in the multi-well plate may be realized by using the two-dimensional displacement table controlled by the upper computer 20.
Fig. 6 is a schematic structural diagram of a full spectrum fluorescence lifetime rapid imaging system in embodiment 4 of the present invention. The imaging system comprises the full-spectrum fluorescence lifetime rapid measuring device shown in fig. 3, and can realize full-spectrum fluorescence lifetime rapid imaging of a sample.
As shown in fig. 6, the excitation light emitted by the pulse laser 9 enters the fluorescence excitation and collection assembly 10-19, is reflected when reaching the dichroic mirror 10, enters the two-dimensional galvanometer system consisting of the X-axis motor and the reflecting mirror 15 thereof and the Y-axis motor and the reflecting mirror 16 thereof through the second reflecting mirror 11, sequentially passes through the scanning lens 17 and the sleeve lens 18, is converged to the sample surface in the sample cell 13 through the first focusing lens 12, completes the excitation of the sample, returns the fluorescence path emitted by the sample, is transmitted through the dichroic mirror 10, enters the full-spectrum fluorescence lifetime rapid measuring device 1-8 through the second focusing lens 14 and the small hole 19, performs the light splitting and spot shaping processing of fluorescence through the optical system, and performs the rapid detection and transmission of fluorescence information through the detection circuit system. The signal processing system 8 monitors the transmitted digital signal and the pulse synchronous signal of the pulse laser 9 at the same time, and transmits the information to the upper computer 20, and the upper computer 20 completes statistics and further calculation of the fluorescence information of the sample to obtain the full spectrum fluorescence lifetime information of the sample.
Preferably, in the fluorescence excitation and collection assemblies 10-19, the X-axis motor and its reflecting mirror 15, the Y-axis motor and its reflecting mirror 16, the scanning lens 17, the sleeve lens 18 and the small hole 19 are confocal laser scanning microscopic imaging components, and the two-dimensional galvanometer system can be regulated and controlled by the upper computer 20, so that the full-spectrum fluorescence lifetime rapid imaging of the sample is realized.
It will be apparent to those skilled in the art that the above embodiments are provided for illustration only and not for limitation of the invention, and that variations and modifications of the above described embodiments are intended to fall within the scope of the claims of the invention as long as they fall within the true spirit of the invention.
Claims (8)
1. A full-spectrum fluorescence lifetime rapid measurement device is characterized by comprising an optical system and a detection circuit system,
the optical system sequentially comprises the following components along an optical path:
a collimation assembly for collimating the fluorescence into an approximately parallel beam;
the light splitting component is used for splitting fluorescence with different wavelengths in one-dimensional direction, namely X direction;
the optical shaping component is used for shaping the fluorescent light spots with Gaussian light intensity distribution into light spots with uniform light intensity distribution, namely rectangular fluorescent light spots, in the other dimension direction, namely the Y direction;
the detection circuitry includes a multi-channel detector and a signal processing system and performs the steps of:
receiving the rectangular fluorescent light spots processed by the optical system, wherein the rectangular fluorescent light spots are separated in wavelength in one dimension, namely the X direction, have uniform light intensity distribution in the other dimension, namely the Y direction, and photon counting is carried out on each of a plurality of pixel units in the X direction and the Y direction of the multichannel detector by converting optical signals into electric signals,
obtaining the time difference between the arrival of the electric pulse synchronous signal and the fluorescent signal of each pixel unit,
obtaining a cumulative histogram of photon numbers of each pixel unit along with time channels according to photon counts of each pixel unit and time differences between arrival of corresponding electric pulse synchronous signals and fluorescence signals of each pixel unit,
each pixel unit in a plurality of pixel units in the Y direction of the multichannel detector receives fluorescence of the same wavelength, and the fluorescence lifetime attenuation histogram under the same wavelength is rapidly obtained by processing photon number cumulative histogram data in the Y direction in parallel; different pixel units of the multichannel detector in the X direction correspond to different fluorescence wavelengths, so that a full-spectrum fluorescence lifetime attenuation histogram, namely full-spectrum fluorescence lifetime information, is further and rapidly obtained.
2. The rapid full-spectrum fluorescence lifetime measurement device of claim 1, wherein in the optical system, the light splitting component is a dispersive element, and the dispersive element is a grating or a prism.
3. The rapid full-spectrum fluorescence lifetime measurement device of claim 1, wherein said optical shaping assembly is positioned after said beam-splitting assembly or before said beam-splitting assembly in the optical path of said optical system.
4. A full spectrum fluorescence lifetime rapid measurement device in accordance with claim 3, wherein the optical shaping element is a powell lens before the light splitting element.
5. A full spectrum fluorescence lifetime rapid measurement device according to claim 3, wherein the optical shaping element is a plano-concave cylindrical mirror and a powell lens, or a plano-convex cylindrical mirror and a powell lens, when the optical shaping element is behind the beam splitting element.
6. The rapid full-spectrum fluorescence lifetime measurement device of claim 1, wherein the multi-channel detector is a planar array type multi-pixel single photon detector or a two-dimensional combination of a plurality of single pixel single photon detectors.
7. The full spectrum fluorescence lifetime rapid measurement device of claim 1, wherein the multi-channel detector is configured to receive the rectangular light spots, to effect photon counting on each pixel cell, wherein the signal processing system is configured to obtain a time difference between arrival of the electrical pulse synchronization signal and the fluorescence signal for each pixel cell, and to obtain a cumulative histogram of photon numbers per pixel cell over time channels; or the multi-channel detector is configured to effect photon counting per pixel cell and to obtain a time difference between arrival of the electrical pulse synchronization signal and the fluorescence signal per pixel cell, the signal processing system being configured to obtain a cumulative histogram of photon counts per pixel cell over time channels.
8. The full-spectrum fluorescence lifetime rapid measurement device of claim 1, wherein the measurement device can measure fluorescence of the same wavelength without the beam-splitting component, each of the plurality of pixel units in the Y direction of the multi-channel detector receives fluorescence of the same wavelength, and the fluorescence lifetime attenuation histogram at the same wavelength is rapidly obtained by processing photon number cumulative histogram data in the Y direction in parallel.
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