A fully automated high-throughput spatiotemporal resolved fluorescence spectroscopic imaging system and method

Abstract

The application discloses a kind of full-automatic high-throughput space-time resolution fluorescence spectrum imaging system and method, it is related to fluorescence signal spectrum field, including: workbench, sample stage, objective, for preloading sample stage with sample and placing it on workbench on mechanical arm sample inlet module, for laser beam to the output module of picosecond pulse laser of objective, for simultaneously collecting the spectral data and dynamic data of fluorescence signal generated after sample is focused by laser beam fluorescence signal detection module and control module;Sample stage and objective are sequentially spaced apart on workbench along the vertical direction from top to bottom;Mechanical arm sample inlet module, picosecond pulse laser output module and fluorescence signal detection module are respectively connected with control module communication.This application can realize the simultaneous acquisition of spectral data and dynamic data of fluorescence signal automatically.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of fluorescence signal spectrum technology, and in particular to a full-automatic high-throughput time-space resolution fluorescence spectrum imaging system and method. BACKGROUND

[0002] Steady-state fluorescence signal spectrum and time-resolved fluorescence signal spectrum are commonly used characterization means in low-dimensional material testing. Steady-state fluorescence signal spectrum can directly correlate information such as band gap and defect state density of the material by recording the light excitation under the luminescence peak position, intensity and line width. Time-resolved fluorescence signal spectrum can track the formation, diffusion and annihilation of excited state species, which is of great significance for the design and research of optoelectronic devices based on these materials.

[0003] With the continuous emergence of low-dimensional materials such as graphene, transition metal sulfide and two-dimensional perovskite materials, the research on these materials is shifting from "single verification" to "million-level array screening". The traditional fluorescence signal spectrum test relies on manual sampling and cannot be operated continuously for 24 hours with high throughput. In addition, the parameters need to be manually set and adjusted during the test, so there is a certain error. Such a test method cannot meet the current demand for a large number of samples. In addition, commercial fluorescence spectrometers need different light sources and optical paths to test steady-state fluorescence signal spectrum and time-resolved fluorescence signal spectrum, so they are tested separately, which complicates the instrument and the testing process to some extent. SUMMARY

[0004] To solve the technical problems in the background art, the present application provides a full-automatic high-throughput time-space resolution fluorescence spectrum imaging system and method.

[0005] In a first aspect, the present application provides a full-automatic high-throughput time-space resolution fluorescence spectrum imaging system, comprising: a workbench, a sample stage, an objective lens, a mechanical arm sampling module, a picosecond pulse laser output module, a fluorescence signal detection module and a control module.

[0006] The sample stage and the objective lens are sequentially and spacedly arranged in a vertical direction from top to bottom on the workbench.

[0007] The mechanical arm sampling module is used to grab the sample stage preloaded with samples and place it on the workbench.

[0008] The picosecond pulse laser output module is used to output a laser beam to the objective lens.

[0009] The objective lens is used to focus the laser beam on the sample of the sample stage.

[0010] The fluorescence signal detection module is used to simultaneously collect the spectral data and kinetic data of the fluorescence signal generated by the sample after being focused by the laser beam.

[0011] The mechanical arm sample feeding module, the picosecond pulse laser output module and the fluorescence signal detection module are in communication connection with the control module, and the control module is used for controlling the mechanical arm sample feeding module, the picosecond pulse laser output module and the fluorescence signal detection module to coordinate actions, so as to realize simultaneous collection of spectral data and kinetic data of the fluorescence signal.

[0012] Preferably, an electric focusing module is further included, the electric focusing module is installed between the sample stage and the workbench, the electric focusing module is used for driving the objective lens to approach or move away from the sample stage, and the electric focusing module is in communication connection with the control module.

[0013] The electric focusing module is in communication connection with the control module, and the control module is used for controlling the mechanical arm sample feeding module, the electric focusing module, the picosecond pulse laser output module and the fluorescence signal detection module to coordinate actions, so as to realize simultaneous collection of spectral data and kinetic data of the fluorescence signal.

[0014] Preferably, the electric focusing module is provided with an electric focusing module controller, and the electric focusing module controller is in communication connection with the control module.

[0015] Preferably, in the coordinated working process, the control module controls the picosecond pulse laser output module to output a laser beam to the objective lens, the laser beam is focused on a sample on the sample stage, and the sample generates a fluorescence signal after being irradiated by the laser beam.

[0016] The control module controls the fluorescence signal detection module to collect spectral data of the fluorescence signal in real time, and simultaneously drives the objective lens to approach or move away from the sample stage through the electric focusing module.

[0017] According to the collected spectral data of the fluorescence signal, a target intensity of the spectral data is determined.

[0018] The electric focusing module is controlled to act until the intensity of the spectral data reaches the target intensity.

[0019] When the intensity of the spectral data is the target intensity, the electric focusing module is controlled to stop acting, and the fluorescence signal detection module is controlled to simultaneously collect spectral data and kinetic data of the fluorescence signal generated by the sample after being focused by the laser beam.

[0020] Preferably, according to the collected spectral data of the fluorescence signal, the target intensity of the spectral data is determined, and specifically includes:

[0021] According to the collected spectral data of the fluorescence signal, it is judged whether the intensity of the spectral data gradually increases as the objective lens approaches or moves away from the sample stage.

[0022] If yes, the electric focusing module is controlled to act to continue driving the objective lens to approach or move away from the sample stage until the intensity of the spectral data starts to decrease.

[0023] If not, the electric focusing module is controlled to drive the objective lens to move away from or close to the sample stage, so that the intensity of the spectral data first increases and then decreases;

[0024] The maximum value of the intensity of the spectral data is taken as the target intensity of the spectral data.

[0025] Preferably, an electric displacement stage is further included, which is installed between the sample stage and the workbench, is used to carry the sample stage and drive the sample stage to move along the X direction and the Y direction of the horizontal plane, and is in communication connection with the control module.

[0026] Preferably, the electric displacement stage is configured with an electric displacement stage controller, which is in communication connection with the control module.

[0027] Preferably, the picosecond pulsed laser output module includes a laser controller, a picosecond laser, a first optical fiber, a first lens and a first filter, a first mirror and a second mirror;

[0028] The control module, the laser controller and the picosecond laser are in communication connection in sequence, the laser generated by the picosecond laser is emitted by the first optical fiber and passes through the first lens, the first filter, the first mirror and the second mirror in sequence, and is output to the objective lens.

[0029] Preferably, the fluorescence signal detection module includes a condenser lens, a second filter, a beam splitting optical fiber, a single photon detector and a fluorescence spectrometer, and the single photon detector and the fluorescence spectrometer are in communication connection with the control module respectively;

[0030] The condenser lens is used to focus the fluorescence signal and the scattered light generated after the sample laser beam is focused; the second filter is used to filter the focused light to retain the fluorescence signal;

[0031] The beam splitting optical fiber is used to collect the fluorescence signal and output the collected fluorescence signal to the single photon detector and the fluorescence spectrometer respectively;

[0032] The single photon detector is used to collect dynamic data of the fluorescence signal;

[0033] The fluorescence spectrometer is used to collect spectral data of the fluorescence signal.

[0034] In a second aspect, the present application further provides a full-automatic high-throughput time-space resolution fluorescence spectrum imaging method, which is applied to the full-automatic high-throughput time-space resolution fluorescence spectrum imaging system in any one of the first aspect and includes:

[0035] The control module controls the mechanical arm sample feeding module to grab the sample stage pre-loaded with the sample and place it on the workbench;

[0036] The control module controls the picosecond pulse laser output module to output a laser beam to the objective lens, and the laser beam is focused on a sample on the sample table through the objective lens.

[0037] The control module controls the fluorescence signal detection module to simultaneously collect spectral data and kinetic data of the fluorescence signal generated after the sample is focused by the laser beam.

[0038] Preferably, before the control module controls the fluorescence signal detection module to simultaneously collect spectral data and kinetic data of the fluorescence signal generated after the sample is focused by the laser beam, the method further comprises:

[0039] The control module controls the fluorescence signal detection module to collect spectral data of the fluorescence signal in real time, and controls the electric focusing module to drive the objective lens to move towards or away from the sample table;

[0040] According to the collected spectral data of the fluorescence signal, the target intensity of the spectral data is determined.

[0041] The electric focusing module is controlled to act until the intensity of the spectral data is the target intensity.

[0042] When the intensity of the spectral data is the target intensity, the electric focusing module is controlled to stop acting.

[0043] Preferably, according to the collected spectral data of the fluorescence signal, the target intensity of the spectral data is determined, and specifically comprises:

[0044] According to the collected spectral data of the fluorescence signal, it is judged whether the intensity of the spectral data gradually increases as the objective lens moves towards or away from the sample table.

[0045] If yes, the electric focusing module is controlled to act to continue driving the objective lens to move towards or away from the sample table until the intensity of the spectral data starts to decrease.

[0046] If no, the electric focusing module is controlled to act to drive the objective lens to move away from or towards the sample table, so that the intensity of the spectral data first increases and then decreases.

[0047] The maximum value of the intensity of the spectral data is taken as the target intensity of the spectral data.

[0048] In the present application, the proposed full-automatic high-throughput time-space resolution fluorescence spectrum imaging system realizes the simultaneous collection of spectral data and kinetic data of the fluorescence signal through the control module controlling the mechanical arm sample feeding module, the picosecond pulse laser output module and the fluorescence signal detection module to act in coordination, thereby solving the problems that the existing fluorescence spectrometer cannot realize high-throughput continuous operation and separate testing of steady-state fluorescence signal spectrum and time-resolved fluorescence signal spectrum. BRIEF DESCRIPTION OF DRAWINGS

[0049] Figure 1This is a schematic diagram of the structure of a fully automated high-throughput spatiotemporal resolution fluorescence spectroscopy imaging system according to an embodiment of the present invention.

[0050] Figure label:

[0051] 1-Computer; 2-First data transmission line; 3-Laser controller; 4-Second data transmission line

[0052] 5-Picosecond laser; 6-First optical fiber; 7-First lens; 8-First filter

[0053] 9-First reflecting mirror; 10-Second reflecting mirror; 11-Electric focusing module; 12-Objective lens

[0054] 13-Electrically operated stage; 14-Sample stage; 15-Condenser lens; 16-Second filter

[0055] 17-Bundled fiber optic cable; 18-Single-photon detector; 19-Third data transmission line; 20-Fluorescence spectrometer

[0056] 21-Fourth data transmission line; 22-Fifth data transmission line; 23-Robotic arm sample feeding module

[0057] 24 - Sixth data transmission line 25 - Seventh data transmission line

[0058] 26-Electric displacement stage controller; 27-Electric focusing module controller

[0059] 28 - Eighth data transmission line; 29 - Ninth data transmission line Detailed Implementation

[0060] It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other. The present invention will now be described in detail with reference to the accompanying drawings and embodiments.

[0061] Firstly, such as Figure 1 As shown, the present invention proposes a fully automated high-throughput spatiotemporal resolution fluorescence spectroscopy imaging system, comprising: a worktable, a sample stage 14, an objective lens 12, a robotic arm sample feeding module 23, a picosecond pulse laser output module, a fluorescence signal detection module, and a control module;

[0062] The sample stage 14 and the objective lens 12 are arranged at intervals on the worktable in a vertical direction from top to bottom;

[0063] The robotic arm sample feeding module 23 is used to pick up the sample stage 14 pre-loaded with samples and place it on the worktable;

[0064] The picosecond pulse laser output module is used to output a laser beam to objective lens 12;

[0065] The objective lens 12 is used for focusing the laser beam onto the sample on the sample stage 14;

[0066] The fluorescence signal detection module is used for simultaneously collecting the spectral data and the kinetic data of the fluorescence signal generated after the sample is focused by the laser beam;

[0067] The mechanical arm sample feeding module 23, the picosecond pulse laser output module and the fluorescence signal detection module are respectively in communication connection with the control module, and the control module is used for controlling the mechanical arm sample feeding module 23, the picosecond pulse laser output module and the fluorescence signal detection module to coordinate actions, so as to realize the simultaneous collection of the spectral data and the kinetic data of the fluorescence signal.

[0068] The mechanical arm sample feeding module 23 is used for grabbing and placing the sample stage 14 preloaded with the sample on the workbench, so as to realize the automatic sample feeding and avoid the labor-intensive work process of manually searching for the sample and manually switching the sample.

[0069] The control module is used for controlling the mechanical arm sample feeding module 23, the picosecond pulse laser output module and the fluorescence signal detection module to coordinate actions, so as to realize the simultaneous collection of the spectral data and the kinetic data of the fluorescence signal, thereby solving the problems that the existing fluorescence spectrometer 20 cannot realize the high-throughput continuous operation and separate test of the steady-state fluorescence signal spectrum and the time-resolved fluorescence signal spectrum.

[0070] Specifically, the mechanical arm sample feeding module 23 is connected with the control module through the fifth data transmission line 22.

[0071] In one of the embodiments, an electric focusing module 11 is further included, the electric focusing module 11 is installed between the sample stage 14 and the workbench, the electric focusing module 11 is used for driving the objective lens 12 to move close to or away from the sample stage 14, and the electric focusing module 11 is in communication connection with the control module.

[0072] The electric focusing module 11 is in communication connection with the control module, and the control module is used for controlling the mechanical arm sample feeding module 23, the electric focusing module 11, the picosecond pulse laser output module and the fluorescence signal detection module to coordinate actions, so as to realize the simultaneous collection of the spectral data and the kinetic data of the fluorescence signal.

[0073] The electric focusing module 11 in the embodiment is configured with an electric focusing module controller 27, and the electric focusing module controller 27 is in communication connection with the control module.

[0074] Specifically, the electric focusing module 11 is connected with the electric focusing module controller 27 through the ninth data transmission line 29, and the electric focusing module controller 27 is connected with the control module through the seventh data transmission line 25.

[0075] It should be understood that the smaller the laser spot is, the greater the intensity of the spectral data is, and the more minute regions can be detected. In the embodiment, the electric focusing module 11 is controlled to make the laser spot in the spot image reach the minimum, so that the signal of the minute region meeting the preset requirement can be detected.

[0076] In a further embodiment, in the coordination process, the control module controls the picosecond pulse laser output module to output a laser beam to the objective lens 12, and the laser beam is focused on the sample on the sample stage 14. After the sample is irradiated by the laser beam, a fluorescence signal is generated;

[0077] The control module controls the fluorescence signal detection module to collect the spectral data of the fluorescence signal in real time, and controls the electric focusing module 11 to drive the objective lens 12 to move towards or away from the sample stage 14;

[0078] According to the collected spectral data of the fluorescence signal, the target intensity of the spectral data is determined;

[0079] The electric focusing module 11 is controlled to act until the intensity of the spectral data is the target intensity;

[0080] When the intensity of the spectral data is the target intensity, the electric focusing module 11 is controlled to stop acting, and the fluorescence signal detection module is controlled to collect the spectral data and the kinetic data of the fluorescence signal generated after the sample is focused by the laser at the same time.

[0081] In a further embodiment, according to the collected spectral data of the fluorescence signal, the target intensity of the spectral data is determined, specifically including:

[0082] According to the collected spectral data of the fluorescence signal, it is determined whether the intensity of the spectral data gradually increases as the objective lens 12 moves towards or away from the sample stage 14;

[0083] If yes, the electric focusing module 11 is controlled to act to continue driving the objective lens 12 to move towards or away from the sample stage 14 until the intensity of the spectral data begins to decrease;

[0084] If no, the electric focusing module 11 is controlled to act to drive the objective lens 12 to move away from or towards the sample stage 14, so that the intensity of the spectral data first increases and then decreases;

[0085] The maximum value of the intensity of the spectral data is taken as the target intensity of the spectral data.

[0086] It should be understood that the sample table 14 in the embodiment is used to place a plurality of samples, and the pre-excitation areas of each sample are not the same. In order to realize automatic switching of the samples and adjustment of the positions of the pre-excitation areas of each sample after switching, in one of the embodiments, an electric displacement table 13 is further included, which is installed between the sample table 14 and the workbench, is used to carry the sample table 14 and drive the sample table 14 to move along the X direction and the Y direction of the horizontal plane, and is in communication connection with the control module.

[0087] The electric displacement table 13 in the embodiment is configured with an electric displacement table controller 26, which is in communication connection with the control module.

[0088] Specifically, the electric displacement table 13 is connected with the electric displacement table controller 26 through an eighth data transmission line 28, and the electric displacement table controller 26 is in communication connection with the control module through the sixth data transmission line 24.

[0089] In one of the embodiments, the picosecond pulsed laser output module includes a laser controller 3, a picosecond laser 5, a first optical fiber 6, a first lens 7, a first filter 8, a first mirror 9 and a second mirror 10.

[0090] The control module, the laser controller 3 and the picosecond laser 5 are sequentially in communication connection, and the laser generated by the picosecond laser 5 is output onto the objective lens 12 after being emitted by the first optical fiber 6 and sequentially passing through the first lens 7, the first filter 8, the first mirror 9 and the second mirror 10.

[0091] The first filter 8 in the embodiment is a neutral density filter.

[0092] Specifically, the control module is connected with the laser controller 3 through a first data transmission line 2, and the laser controller 3 and the picosecond laser 5 are connected through a second data transmission line 4.

[0093] In one of the embodiments, the fluorescence signal detection module includes a condenser lens 15, a second filter 16, a beam splitting optical fiber 17, a single photon detector 18 and a fluorescence spectrometer 20, and the single photon detector 18 and the fluorescence spectrometer 20 are respectively in communication connection with the control module.

[0094] The condenser lens 15 is used to focus the fluorescence signal and the scattered light generated after the sample is focused by the laser beam; the second filter is used to filter out the laser and the white light in the focused light, so as to retain the fluorescence signal.

[0095] The beam splitting optical fiber 17 is used to collect the fluorescence signal and output the collected fluorescence signal to the single photon detector 18 and the fluorescence spectrometer 20 respectively.

[0096] The single photon detector 18 is used for dynamic data acquisition of the fluorescent signal;

[0097] The fluorescence spectrometer 20 is used for spectral data acquisition of the fluorescent signal.

[0098] The embodiment collects and focuses the fluorescent signal emitted by the sample after excitation by the condenser lens 15, filters the light of specific wavelength by the second filter 16 to reduce interference on detection, and uses the beam splitting fiber 17 to conduct the fluorescent signal to the single photon detector 18 and the fluorescence spectrometer 20 at the same time, so that the dynamic data and the spectral data of the fluorescent signal of the sample can be obtained at the same time, and the test efficiency and the test accuracy are greatly improved.

[0099] It should be understood that the condenser lens 15 is arranged directly above the sample stage 14 to collect and focus the fluorescent signal emitted by the sample after excitation, the second filter 16 is arranged directly above the condenser lens 15, and the detection end of the beam splitting fiber 17 is located directly above the second filter 16 to receive the fluorescent light transmitted by the second filter 16.

[0100] Specifically, the single photon detector 18 is connected with the control module through the third data transmission line 19, and the fluorescence spectrometer 20 is connected with the control module through the fourth data transmission line 21.

[0101] In one specific embodiment, the control module is a computer 1.

[0102] In a second aspect, the present application further provides a full-automatic high-throughput time-space resolution fluorescence spectrum imaging method, which is applied to the full-automatic high-throughput time-space resolution fluorescence spectrum imaging system in any one of the first aspect, and comprises the following steps:

[0103] The control module controls the mechanical arm sample feeding module 23 to grab the sample stage 14 preloaded with the sample and place it on the workbench;

[0104] The control module controls the picosecond pulse laser output module to output a laser beam to the objective lens 12, and the laser beam is focused on the sample on the sample stage 14 through the objective lens 12;

[0105] The control module controls the fluorescent signal detection module to simultaneously collect the spectral data and the dynamic data of the fluorescent signal generated by the sample after being focused by the laser beam.

[0106] In one embodiment, before the control module controls the fluorescent signal detection module to simultaneously collect the spectral data and the dynamic data of the fluorescent signal generated by the sample after being focused by the laser beam, the method further comprises the following steps:

[0107] The control module controls the fluorescent signal detection module to collect the spectral data of the fluorescent signal in real time, and simultaneously controls the electric focusing module 11 to drive the objective lens 12 to move towards or away from the sample stage 14;

[0108] determining a target intensity of the spectral data according to the spectral data of the fluorescence signal;

[0109] controlling the electric focusing module 11 to act until the intensity of the spectral data reaches the target intensity;

[0110] controlling the electric focusing module 11 to stop acting when the intensity of the spectral data is the target intensity.

[0111] In a further embodiment, the target intensity of the spectral data is determined according to the spectral data of the fluorescence signal, specifically comprising:

[0112] determining whether the intensity of the spectral data gradually increases as the objective lens 12 approaches or moves away from the sample stage 14 according to the spectral data of the fluorescence signal;

[0113] if yes, controlling the electric focusing module 11 to act to continue driving the objective lens 12 to approach or move away from the sample stage 14 until the intensity of the spectral data begins to decrease;

[0114] if no, controlling the electric focusing module 11 to act to drive the objective lens 12 to move away or approach the sample stage 14 so that the intensity of the spectral data first increases and then decreases;

[0115] taking the maximum value of the intensity of the spectral data as the target intensity of the spectral data.

[0116] The application will be described in detail below with reference to specific embodiments.

[0117] Embodiment 1

[0118] The embodiment discloses a full-automatic high-throughput spatiotemporal resolution fluorescence spectral imaging system, comprising a workbench, a sample stage 14, an electric focusing module 11, an objective lens 12, a mechanical arm sample feeding module 23, a picosecond pulse laser output module, a fluorescence signal detection module, and a computer 1.

[0119] The sample stage 14 and the objective lens 12 are sequentially and spacedly arranged in a vertical direction from top to bottom on the workbench; the electric focusing module 11 is installed between the sample stage 14 and the workbench, and the electric focusing module 11 is used to drive the objective lens 12 to approach or move away from the sample stage 14;

[0120] The mechanical arm sample feeding module 23 is used to grab and place the sample stage 14 preloaded with a sample on the workbench;

[0121] The picosecond pulse laser output module is used to output a laser beam to the objective lens 12;

[0122] The objective lens 12 is used to focus the laser beam on a sample of the sample stage 14;

[0123] The fluorescence signal detection module is used for simultaneously collecting spectral data and dynamic data of the fluorescence signal generated by the sample after being focused by the laser beam;

[0124] The electric focusing module 11, the mechanical arm sample feeding module 23, the picosecond pulse laser output module and the fluorescence signal detection module are respectively in communication connection with the computer 1, and the computer 1 is used for controlling the mechanical arm sample feeding module 23, the electric focusing module 11, the picosecond pulse laser output module and the fluorescence signal detection module to coordinate actions, so as to realize simultaneous collection of spectral data and dynamic data of the fluorescence signal.

[0125] In the coordination process, the computer 1 controls the picosecond pulse laser output module to output a laser beam to the objective lens 12, the laser beam is focused on the sample on the sample stage 14, and the sample generates a fluorescence signal after being irradiated by the laser beam;

[0126] The computer 1 controls the fluorescence signal detection module to collect the spectral data of the fluorescence signal in real time, and simultaneously drives the objective lens 12 to move towards or away from the sample stage 14 through the electric focusing module 11;

[0127] According to the collected spectral data of the fluorescence signal, it is judged whether the intensity of the spectral data gradually increases with the objective lens 12 moving towards or away from the sample stage 14;

[0128] If yes, the electric focusing module 11 is controlled to act to continue driving the objective lens 12 to move towards or away from the sample stage 14 until the intensity of the spectral data begins to decrease;

[0129] If no, the electric focusing module 11 is controlled to act to drive the objective lens 12 to move away from or towards the sample stage 14, so that the intensity of the spectral data first increases and then decreases;

[0130] The maximum value of the intensity of the spectral data is taken as a target intensity of the spectral data;

[0131] The electric focusing module 11 is controlled to act until the intensity of the spectral data reaches the target intensity;

[0132] When the intensity of the spectral data is the target intensity, the electric focusing module 11 is controlled to stop acting, and the fluorescence signal detection module is controlled to simultaneously collect the spectral data and dynamic data of the fluorescence signal generated by the sample after being focused by the laser.

[0133] The fluorescence signal detection module in the embodiment includes a condenser lens 15, a second filter 16, a beam splitting optical fiber 17, a single photon detector 18 and a fluorescence spectrometer 20, and the single photon detector 18 and the fluorescence spectrometer 20 are respectively in communication connection with the computer 1;

[0134] The picosecond pulse laser output module in the embodiment comprises a laser controller 3, a picosecond laser 5, a first optical fiber 6, a first lens 7 and a neutral density filter, a first mirror 9 and a second mirror 10.

[0135] The motorized displacement stage 13 in the embodiment is configured with a motorized displacement stage controller 26, which is in communication connection with the computer 1. The motorized focusing module 11 in the embodiment is configured with a motorized focusing module controller 27, which is in communication connection with the computer 1.

[0136] The above is only the preferred specific implementation of the present application, but the protection scope of the present application is not limited to this, any person skilled in the art in the technical range disclosed by the present application, according to the technical scheme and the inventive concept of the present application, equivalent replacement or change, should be covered in the protection scope of the present application.

Claims

1. A fully automated high-throughput spatiotemporal resolution fluorescence spectroscopy imaging system, characterized in that, include: Worktable, sample stage, objective lens, robotic arm sample feeding module, picosecond pulse laser output module, fluorescence signal detection module, and control module; The fluorescence signal detection module includes a focusing lens, a second filter, a beam splitter fiber, a single-photon detector, and a fluorescence spectrometer. The single-photon detector and the fluorescence spectrometer are respectively connected to the control module. The sample stage and objective lens are arranged at intervals along the vertical direction from top to bottom on the worktable; The robotic arm sample feeding module is used to grasp the sample stage pre-loaded with samples and place it on the worktable; The picosecond pulsed laser output module is used to output a laser beam to the objective lens; The objective lens is used to focus the laser beam onto the sample on the sample stage; The condenser lens is used to focus the fluorescence signal and scattered light generated after the sample laser beam is focused; the second filter is used to filter the focused light to retain the fluorescence signal. The fiber optic bundle is used to collect fluorescence signals and outputs the collected fluorescence signals to a single-photon detector and a fluorescence spectrometer, respectively. Single-photon detectors are used for acquiring dynamic data of fluorescence signals; A fluorescence spectrometer is used to acquire spectral data of fluorescence signals; The control module is used to coordinate the actions of the robotic arm sample feeding module, picosecond pulse laser output module, and fluorescence signal detection module to achieve simultaneous acquisition of spectral and kinetic data of fluorescence signals.

2. The fully automated high-throughput spatiotemporal resolution fluorescence spectroscopy imaging system according to claim 1, characterized in that, It also includes an electric focusing module, which is installed between the sample stage and the work stage. The electric focusing module is used to drive the objective lens to move closer to or away from the sample stage. The electric focusing module is connected to the control module. The control module is used to coordinate the actions of the robotic arm sample feeding module, electric focusing module, picosecond pulse laser output module and fluorescence signal detection module to achieve simultaneous acquisition of spectral and kinetic data of fluorescence signals.

3. The fully automated high-throughput spatiotemporal resolution fluorescence spectroscopy imaging system according to claim 2, characterized in that, The electric focusing module is equipped with an electric focusing module controller, which is connected to the control module for communication.

4. The fully automated high-throughput spatiotemporal resolution fluorescence spectroscopy imaging system according to claim 2, characterized in that, During the coordinated operation, the control module controls the picosecond pulse laser output module to output a laser beam to the objective lens. The laser beam is focused onto the sample on the sample stage, and the sample generates a fluorescence signal after being irradiated by the laser beam. The control module controls the fluorescence signal detection module to acquire the spectral data of the fluorescence signal in real time, and at the same time drives the objective lens to move closer to or away from the sample stage through the electric focusing module; Based on the spectral data of the collected fluorescence signals, determine the target intensity of the spectral data; Control the electric focusing module to operate until the intensity of the spectral data reaches the target intensity; When the intensity of the spectral data reaches the target intensity, the electric focusing module stops operating, and the fluorescence signal detection module simultaneously acquires the spectral and kinetic data of the fluorescence signal generated after the sample is focused by the laser.

5. The fully automated high-throughput spatiotemporal resolution fluorescence spectroscopy imaging system according to claim 4, characterized in that, Based on the spectral data of the acquired fluorescence signals, the target intensity of the spectral data is determined, specifically including: Based on the spectral data of the acquired fluorescence signal, determine whether the intensity of the spectral data gradually increases as the objective lens moves closer to or further away from the sample stage; If so, control the motorized focusing module to continue driving the objective lens closer to or further away from the sample stage until the intensity of the spectral data begins to decrease; If not, the motorized focusing module is controlled to move the objective lens away from or closer to the sample stage, causing the intensity of the spectral data to first increase and then decrease. The maximum intensity of the spectral data is taken as the target intensity of the spectral data.

6. The fully automated high-throughput spatiotemporal resolution fluorescence spectroscopy imaging system according to claim 1, characterized in that, It also includes an electric displacement stage, which is installed between the sample stage and the worktable. The electric displacement stage is used to support the sample stage and drive the sample stage to move along the X and Y directions of the horizontal plane. The electric displacement stage is connected to the control module.

7. The fully automated high-throughput spatiotemporal resolution fluorescence spectroscopy imaging system according to claim 6, characterized in that, The electric displacement table is equipped with an electric displacement table controller, which is connected to the control module.

8. The fully automated high-throughput spatiotemporal resolution fluorescence spectroscopy imaging system according to claim 1, characterized in that, The picosecond pulsed laser output module includes a laser controller, a picosecond laser, a first optical fiber, a first lens, a first filter, a first reflector, and a second reflector. The control module, laser controller, and picosecond laser are connected in sequence. The laser generated by the picosecond laser is emitted from the first optical fiber and passes through the first lens, the first filter, the first reflector, and the second reflector in sequence before being output to the objective lens.

9. A fully automated high-throughput spatiotemporal resolution fluorescence spectroscopy imaging method, applied to the fully automated high-throughput spatiotemporal resolution fluorescence spectroscopy imaging system according to any one of claims 1-8, characterized in that, include: The control module controls the robotic arm's sample feeding module to grab the sample stage pre-loaded with the sample and place it on the worktable; The control module controls the picosecond pulse laser output module to output a laser beam to the objective lens, and the laser beam is focused onto the sample on the sample stage by the objective lens; The control module controls the fluorescence signal detection module to simultaneously acquire the spectral and kinetic data of the fluorescence signal generated after the sample is focused by the laser beam.

10. The fully automated high-throughput spatiotemporal resolution fluorescence spectroscopy imaging method according to claim 9, characterized in that, Before the control module controls the fluorescence signal detection module to simultaneously acquire the spectral and kinetic data of the fluorescence signal generated after the sample is focused by the laser beam, the following is also included: The control module controls the fluorescence signal detection module to acquire the spectral data of the fluorescence signal in real time, and at the same time controls the electric focusing module to drive the objective lens closer to or away from the sample stage; Based on the spectral data of the collected fluorescence signals, determine the target intensity of the spectral data; Control the electric focusing module to operate until the intensity of the spectral data reaches the target intensity; When the intensity of the spectral data reaches the target intensity, the electric focusing module is controlled to stop operating.

11. The fully automated high-throughput spatiotemporal resolution fluorescence spectroscopy imaging method according to claim 10, characterized in that, Based on the spectral data of the acquired fluorescence signals, the target intensity of the spectral data is determined, specifically including: Based on the spectral data of the acquired fluorescence signal, determine whether the intensity of the spectral data gradually increases as the objective lens moves closer to or further away from the sample stage; If so, control the motorized focusing module to continue driving the objective lens closer to or further away from the sample stage until the intensity of the spectral data begins to decrease; If not, the motorized focusing module is controlled to move the objective lens away from or closer to the sample stage, causing the intensity of the spectral data to first increase and then decrease. The maximum intensity of the spectral data is taken as the target intensity of the spectral data.

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