Spectrum system, device and control method thereof

Abstract

The invention relates to a spectrum system comprising a light source which continuously emits an initial light beam with a certain wavelength; the objective table is used for bearing a sample which is irradiated by the initial light beam to excite a fluorescence signal, and the fluorescence signal forms a fluorescence light beam; the first objective lens has a long working distance and is used for amplifying the fluorescent light beam to form a microscopic light beam; the second objective lens amplifies the microscopic light beam to form an amplified light beam; the transmission optical fiber couples the amplified light beam into the transmission optical fiber to form a detection light beam; the shunt switching device is used for dividing the detection light beam into a steady-state detection light beam and a transient-state detection light beam and controlling output channels of the steady-state detection light beam and the transient-state detection light beam; the steady-state detector is used for receiving the steady-state detection light beam and detecting to obtain a sample fluorescence steady-state signal; and the transient detector is used for receiving the transient detection light beam and detecting to obtain a sample fluorescence transient signal. By adopting the spectrum system disclosed by the invention, the fluorescence steady state signal and the transient state signal of the sample are simultaneously detected in one system, and the laser utilization rate and the signal collection efficiency are improved.

Description

Technical Field

[0001] The present invention relates to the field of spectral applications, and in particular to a spectral system. Background Art

[0002] As the demand for detection in various industries grows, the non-contact and lossless characteristics of the spectral system for detecting samples make it one of the most important detection methods at present. The spectral system mainly obtains the corresponding parameters of the sample by judging the steady state or transient state of its own light signal, and detects the molecular structure and the intermolecular interaction process based on this. In the process of absorbing ultraviolet and visible electromagnetic radiation, the molecules are stimulated to transition to an excited state, and most molecules emit energy in the form of heat through collisions with other molecules; while a small number of molecules emit this energy in the form of light, and the wavelength of the emitted light is different from the wavelength of the radiation absorbed by the molecules. This process is called photoluminescence. The fluorescence of molecules is a type of photoluminescence. The spectral system obtains steady-state information by detecting the fluorescence spectrum and obtains transient information by detecting the fluorescence lifetime, thereby clearly observing the structure and action process between molecules.

[0003] Traditional spectral systems are divided into independent steady-state spectral systems and transient spectral systems, which can detect the steady-state fluorescence signal and transient signal of the sample respectively. However, the independent steady-state spectral system and transient spectral system cannot fully detect the molecular information of the sample. Therefore, the existing spectral system combines the steady-state spectral system and the transient spectral system together to simultaneously detect the complete composition and molecular structure information of the sample.

[0004] However, the existing spectral system simply combines the steady-state spectral system and the transient spectral system, and only shares some devices. The two actually still work in their own optical path structures, which has low energy utilization and signal collection efficiency. The overall structure required is complicated, the optical path devices are repeated and numerous, and the cost is high. Summary of the invention

[0005] Based on this, the purpose of the present invention is to provide a spectral system that can simultaneously detect sample fluorescence steady-state signals and transient signals in a simple optical path structure, improve laser utilization and signal collection efficiency, and greatly reduce system costs.

[0006] A spectroscopy system comprising

[0007] A light source that continuously emits an initial beam of light of a certain wavelength;

[0008] A stage, carrying a sample that is irradiated by the initial light beam and excites a fluorescent signal, wherein the fluorescent signal forms a fluorescent light beam;

[0009] A first objective lens, which amplifies the fluorescent light beam to form a microscopic light beam;

[0010] a second objective lens, which amplifies the microscopic light beam to form an amplified light beam;

[0011] A transmission optical fiber couples the amplified light beam into the transmission optical fiber to form a detection light beam;

[0012] A branching switching device, which divides the detection beam into a steady-state detection beam and a transient detection beam, and controls the output channels of the steady-state detection beam and the transient detection beam;

[0013] A steady-state detector receives a steady-state detection beam and detects and obtains a steady-state fluorescence signal of the sample;

[0014] A transient detector receives a transient detection light beam and detects a transient fluorescence signal of the sample;

[0015] Wherein, the first objective lens is a long working distance objective lens, so that the initial light beam can be incident on the sample at a large angle.

[0016] Furthermore, the branch switching device is an optical path switch, and the connectivity selection of the optical path can be changed by changing the optical path switch, so that all detection beams are input into the steady-state detector as steady-state detection beams or all detection beams are input into the transient detector as transient detection beams.

[0017] Furthermore, the branch switching device is a beam splitter, and the beam splitter splits the detection beam into a steady-state detection beam and a transient detection beam with the same intensity at an energy splitting ratio of 50 / 50.

[0018] Furthermore, it also includes a beam splitter and an observation device; the beam splitter is arranged between the first objective lens and the second objective lens, and transmits the microscopic light beam with a certain energy ratio to form a light beam to be detected, and reflects the microscopic light beam to form an observation light beam, and the light beam to be detected forms an amplified light beam after passing through the second objective lens, and the observation light beam is received by the observation device; or the beam splitter is arranged between the first objective lens and the second objective lens, and reflects the microscopic light beam with a certain energy ratio to form a light beam to be detected, and transmits the microscopic light beam to form an observation light beam, and the light beam to be detected forms an amplified light beam after passing through the second objective lens, and the observation light beam is received by the observation device; the observation device includes an eyepiece, a camera and a push-pull rod, and the position of the push-pull rod is changed to switch between the eyepiece receiving the observation light beam or the camera receiving the observation light beam.

[0019] Furthermore, it also includes a positioning reflector to reflect the initial light beam to form an excitation light beam, and the excitation light beam is irradiated to the sample surface. The positioning reflector is rotatable so that the excitation light beam is irradiated on the target detection area of ​​the sample.

[0020] Furthermore, it also includes an attenuator and a beam expander. The attenuator is arranged in the emitting direction of the initial light beam, and attenuates the energy of the initial light beam to form an adjusted light beam; the beam expander expands the adjusted light beam to obtain an extended light beam, and expands the beam diameter to reduce the beam energy density. The extended light beam is reflected by the positioning reflector to form an excitation light beam.

[0021] The present invention also provides a spectral device, including the above-mentioned spectral system, an industrial computer, a servo drive, a main control board, multiple servos and multiple stepper motors; the industrial computer is connected to the light source, the steady-state detector, the oscilloscope, the servo drive, the main control board and the observation device, and sends corresponding adjustment instructions to the light source, the steady-state detector, the oscilloscope, the servo drive and the main control board according to known input parameters, and simultaneously receives data from the observation device and displays the picture on the screen of the industrial computer; the servo drive is connected to each servo, and sends corresponding control instructions to control the displacement change of the corresponding servo according to the adjustment instructions of the industrial computer; the main control board is connected to each stepper motor, and sends corresponding control instructions to control the displacement change of the corresponding stepper motor according to the adjustment instructions of the industrial computer.

[0022] Furthermore, the servo includes an attenuator servo, an observation switching servo and a positioning servo; the attenuator servo is connected to the attenuator, and controls the change of the attenuation degree of the attenuator according to the control instruction of the servo driver; the observation switching servo is connected to the push-pull rod of the observation device, and controls the switching of the optical path connection between the eyepiece and the spectrometer and the optical path connection between the camera and the spectrometer in the observation device according to the control instruction of the servo driver; the positioning servo is connected to the positioning reflector, and drives the rotation and movement of the positioning reflector according to the control instruction of the servo driver, so that the position of the spot irradiated by the excitation light beam on the sample surface moves, and controls the center of the spot to be irradiated in the sample target detection area.

[0023] Furthermore, the stepper motor includes a stage coarse adjustment stepper motor and a stage fine adjustment stepper motor; the stage coarse adjustment stepper motor is connected to the stage, and drives the stage to move with coarse steps according to control instructions from the main control board; the stage fine adjustment stepper motor is connected to the stage, and drives the stage to move with fine steps according to control instructions from the main control board.

[0024] Furthermore, it also includes an oscilloscope and a high-voltage controller. The oscilloscope is connected to the steady-state detector and the transient detector respectively, and presents the fluorescent steady-state signal and the transient signal in a clear curve change according to the adjustment instructions of the industrial computer; the high-voltage controller is connected to the PMT detector inside the steady-state detector, and controls the operation of the PMT detector according to the control instructions of the main control board.

[0025] Furthermore, it also includes a backlight servo and a backlight lamp, the backlight servo is connected to the servo driver, and the servo driver controls the backlight servo according to the adjustment instructions of the industrial computer; the backlight servo is connected to the backlight lamp, and the brightness of the backlight lamp is controlled according to the control instructions of the servo driver.

[0026] The present invention also provides a control method for the spectral equipment as described above, wherein appropriate parameters are selected on an industrial computer according to the properties of the sample, and an instruction for steady-state spectrum detection is sent to a steady-state detector. The steady-state detector adjusts its own parameters according to the instruction for steady-state spectrum detection, detects fluorescence steady-state signal information, and inputs the detected steady-state information into a channel of an oscilloscope; the industrial computer sends an adjustment instruction to the oscilloscope, the oscilloscope sets its own parameters according to the adjustment instruction, receives the steady-state information detected by the steady-state spectrometer, and makes a corresponding steady-state curve.

[0027] Furthermore, the industrial computer sends an adjustment instruction to the light source, and the light source turns off the laser output according to the adjustment instruction; the transient detector detects the fluorescent transient signal information, and inputs the detected transient information into another channel of the oscilloscope; the industrial computer sends an adjustment instruction to the oscilloscope, and the oscilloscope sets the average number of times, trigger level and other parameters selected by the user according to the adjustment instruction, and receives the steady-state information detected by the transient detector to make a corresponding transient curve.

[0028] Furthermore, before controlling the spectral device to start detection, the detection accuracy of the spectral device is improved by the following steps:

[0029] The industrial computer issues instructions to the main control board, and the main control board controls the stage stepper motor to move, driving the stage to pop out, and after the sample is placed in the center of the stage, the main control board controls the stage stepper motor to move, driving the stage to retract the sample;

[0030] The industrial computer issues instructions to the servo driver, and the servo driver controls the backlight servo to turn on the backlight to illuminate the sample target detection area;

[0031] Observe the sample through the observation device, control the coarse adjustment of the stage through the stage coarse adjustment stepper motor on the main control board, and control the fine adjustment of the stage through the stage fine adjustment stepper motor; repeatedly operate to focus so that the target detection area of ​​the sample is clearly in the center of the observation field of view;

[0032] Control the microscope backlight servo on the servo control panel to turn off the backlight;

[0033] Sending adjustment instructions to the light source through the industrial computer to generate an initial light beam;

[0034] Sending an adjustment instruction to the steering gear driver through the industrial computer to adjust the attenuator to a certain degree;

[0035] The industrial computer sends an adjustment instruction to the steering gear driver, and the steering gear driver controls the positioning steering gear to drive the positioning reflector to rotate, change the position of the laser spot, and make the spot be located in the center of the observation field of view.

[0036] For better understanding and implementation, the present invention is described in detail below with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 It is a schematic diagram of the structure of the spectrum system of the present invention.

[0038] Figure 2 It is a schematic diagram of the structure of the spectral device of the present invention. DETAILED DESCRIPTION

[0039] The applicant carefully analyzed the existing spectral system and found that the reason for its high cost is that the detection of fluorescence spectrum and the detection of fluorescence lifetime have different configuration requirements for each device in the system. It is still necessary to set up independent optical path structures to meet the configuration requirements for the detection of fluorescence spectrum and the detection of fluorescence lifetime respectively, and multiple lasers and other devices need to be set up, resulting in low energy utilization. For this reason, the applicant tried to set up only one optical path structure, and by changing the configuration of all devices and the parameters related to each other, control the final matching effect to meet the detection requirements of fluorescence spectrum and fluorescence lifetime at the same time, so as to combine the detection of fluorescence spectrum and fluorescence lifetime well into the same optical path to improve the energy utilization and signal collection efficiency of the system.

[0040] Example 1

[0041] See also Figure 1 The spectral system of the present invention includes a light source 10, an attenuator 20, a beam expander 30, a first reflector 41, a second reflector 42, a positioning reflector 43, a focusing lens 50, a stage 60, a first objective lens 71, a second objective lens 72, a spectroscope 80, an observation device 90, a transmission optical fiber 100, a branching device 110, a steady-state detector 120 and a transient detector 130.

[0042] The light source 10 is a 355nm laser, which continuously emits an initial light beam with a wavelength of 355nm for subsequently exciting the fluorescence signal of the sample.

[0043] The attenuator 20 is arranged in the emitting direction of the initial light beam. After the initial light beam passes through the attenuator 20, a certain degree of energy attenuation is generated to form an adjusted light beam. The energy attenuation degree of the attenuator 20 is adjustable, thereby controlling the subsequent laser excitation light intensity at the sample to prevent excessive energy from damaging the sample, thereby controllably adjusting the fluorescence signal intensity.

[0044] The beam expander 30 expands the regulated light beam to obtain an extended light beam, expands the beam diameter to reduce the laser energy density, and further prevents the laser energy from being too high to damage subsequent devices. The beam expander 30 can be a concave lens, which expands the emission angle of the transmitted light beam, so that the beam diameter increases when it reaches the next device; the beam expander 30 can also be a combination of a concave lens and a convex lens, wherein the focal points of the concave lens and the convex lens facing the attenuator 20 coincide, so that the light beam whose emission angle is expanded by the concave lens can be collimated by the convex lens again, preventing energy loss due to large-angle escape during subsequent propagation. Specifically, the beam diameter magnification of the beam expander is 6 times.

[0045] It is understandable that the devices used in the spectral system of the present invention are all low-cost devices. In addition to being able to meet the functional requirements defined in the present invention, there are no requirements for their own strength, heat resistance and maximum threshold of laser irradiation. Therefore, the initial light beam intensity emitted by the light source 10 is low, the attenuator 20 is used to further reduce the laser intensity, and the beam expander 30 is used to reduce the laser energy density to prevent damage to various devices and ensure the normal use of low-cost instruments.

[0046] The first reflector 41, the second reflector 42 and the positioning reflector 43 are arranged so that the expanded light beam can be irradiated on the sample surface on the stage 60, thereby exciting the sample to produce fluorescence. Specifically, since the spectral system has a microscopic arrangement in the longitudinal direction, in order to shorten the transverse axis length of the system, the optical axis where the first reflector 41 and the laser 10 are located is arranged to be parallel to the optical axis where the second reflector 42 and the positioning reflector 43 are located. In particular, the maximum reflection angle of the positioning reflector 43 is 60°, which is caused by the radius of the first objective lens 71. It can be understood by those skilled in the art that the reflector does not necessarily need to be set. It is set to achieve better results, but it is not a necessary condition for the implementation of the spectral system. Even if the reflector is not set at all, the same effect can be achieved by directly irradiating the optical axis of the laser 10 on the sample surface; in addition, the attenuator 20 or the beam expander 30 can be set on the optical axis where the second reflector 42 and the positioning reflector 43 are located. The specific installation position can be based on the actual needs and conditions for the lateral spatial position, wherein the lateral and longitudinal directions are only used to distinguish the propagation direction of the fluorescent signal and its vertical direction, and it is not limited that the direction must be lateral or longitudinal, and the lateral and longitudinal directions can be interchanged uniformly. Under the setting of this embodiment, the laser 10 can be installed together with the stage 60 on the lowest surface of the spectral system, which is convenient for operation and improves anti-interference performance, and the lateral axis length of the spectral system can be compressed by setting multiple reflectors according to actual needs.

[0047] The focusing lens 50 is disposed between the second reflector 42 and the positioning reflector 43 to focus the expanded light beam. The focused expanded light beam is reflected by the positioning reflector 43 to form an excitation light beam. The excitation light beam is precisely focused on the sample surface, and the laser energy can be utilized with maximum efficiency.

[0048] The stage 60 is arranged in the emission direction of the expanded light beam, the sample is placed on the stage 60, the sample receives the expanded light beam, absorbs laser energy and is stimulated to generate fluorescence, and the fluorescence signal is transmitted as a fluorescence light beam in a direction away from the stage 60. Specifically, the wavelength of the fluorescence light beam is greater than 355nm, and the light spot generated by the fluorescence on the sample surface is larger than the light spot size formed when the expanded light beam irradiates the sample surface.

[0049] The first objective lens 71 is arranged in the propagation direction of the fluorescent light beam, and amplifies the fluorescent light beam to obtain a microscopic light beam. The magnification of the first objective lens 71 is 10 times, and the first objective lens 71 has a large working distance to meet the requirements of the expanded light beam irradiating the sample surface at a large angle to obtain a fluorescent signal with a high intensity.

[0050] The beam splitter 80 is disposed between the first objective lens 71 and the second objective lens 72, and reflects the microscopic light beam to obtain the observation light beam, and transmits the microscopic light beam to obtain the light beam to be detected. Specifically, the reflection-transmittance ratio of the beam splitter 80 is not limited and can be set according to actual needs. In another embodiment, the beam splitter 80 transmits the microscopic light beam to obtain the observation light beam, and reflects the microscopic light beam to obtain the light beam to be detected.

[0051] The observation device 90 is arranged in the direction of the observation light beam, receives the observation light beam and displays the fluorescence state of the sample. Specifically, the observation device 90 can be a camera, and the fluorescence state of the sample is indirectly observed by shooting with the camera; the observation device 90 can also be an eyepiece, and the human eye directly observes the fluorescence state of the sample through the eyepiece. The observation device can also include a camera, an eyepiece and a push-pull rod, and the camera or the eyepiece can be switched to access the optical path system through the push-pull rod to realize two different display methods.

[0052] The second objective lens 72 is arranged in the propagation direction of the light beam to be detected, and amplifies the light beam to be detected to obtain an amplified light beam. The magnification of the second objective lens 72 is 10 times.

[0053] The transmission optical fiber 100 is arranged in the emission direction of the amplified light beam, and the amplified light beam is coupled into the transmission optical fiber 100 through the end face of the transmission optical fiber 100 to obtain a detection light beam, further reducing the laser energy to prevent damage to subsequent photosensitive devices, and the spot size of the amplified light beam irradiated on the end face of the transmission optical fiber 100 is smaller than the photosensitive area of ​​the steady-state detector 120 and the transient detector 130, ensuring that all the fluorescent signals collected by the transmission optical fiber 100 can be detected and utilized. Specifically, the transmission optical fiber is an SMA optical fiber with a core of 200μm. It is understandable that the core of the optical fiber is very thin and difficult to reflect in the macroscopic structure diagram, so in Figure 1 Simplified and exaggerated drawings are used to highlight the arrangement of the transmission optical fiber 100 and the position of its end face, rather than to limit the actual size.

[0054] The branching device 110 is disposed in the middle of the transmission optical fiber 100, and divides the detection beam into a steady-state detection beam and a transient detection beam. Specifically, the branching device 110 is a beam splitter with an energy splitting ratio of 50 / 50, which evenly divides the detection beam into a steady-state detection beam and a transient detection beam of equal intensity, so that the spectral system can simultaneously detect the fluorescence steady-state signal and the transient signal, and the measurement efficiency and capacity utilization are high. It can be understood that the energy splitting ratio of the branching device 110 can be changed, and when it is changed, the parameters such as the laser intensity in the system need to be adaptively adjusted.

[0055] The steady-state detector 120 receives the steady-state detection beam and detects a fluorescence steady-state signal. Specifically, the steady-state detector 120 includes a spectrometer, which detects a large-area spectral range through an internal PMT detector to obtain a fluorescence spectrum of the fluorescence signal.

[0056] The transient detector 130 receives the transient detection beam and detects the fluorescent transient signal. Specifically, the transient detector 130 includes an APD detector, which can detect weak single-photon signals and clearly detect the time required for the intensity of the fluorescent signal to decrease to 1 / e of the initial intensity, thereby obtaining the fluorescence lifetime of the fluorescent signal.

[0057] By analyzing the fluorescence steady-state signal and transient signal, the molecular structure and intermolecular interaction process of the sample can be obtained, based on which the composition and change process of the sample can be analyzed. For example, in medical testing, it can distinguish breast cells from tumor cells and monitor the degree of tumor cell pathology.

[0058] The following describes in detail the light beam action process of the spectral system in Example 1 of the present invention.

[0059] The light source 10 emits an initial light beam, which passes through the attenuator 20 to form an energy-attenuated adjustment light beam, which is expanded by the beam expander 30 to form an extended light beam; the extended light beam passes through the first reflector 41 and the second reflector 42 to irradiate the focusing lens 50, is focused by the focusing lens 50 and reflected by the positioning reflector 43 to form an excitation light beam, which is irradiated on the sample surface of the stage 60 to excite the sample to form a fluorescent light beam; the fluorescent light beam passes through the first objective lens 71 to amplify the sample to form a microscopic light beam, which is reflected by the beam splitter 80 to form an observation light beam, and is split The light sheet 80 is transmitted to form a light beam to be detected; the observation light beam enters the observation device 90 and is directly or indirectly observed to obtain the fluorescence state of the sample; the light beam to be detected is amplified after passing through the second objective lens 72 to form an amplified light beam, and the amplified light beam is coupled into the transmission optical fiber 100 through the end face of the transmission optical fiber 100 to form a detection light beam; the detection light beam is evenly divided by the branching device 110 into a steady-state detection light beam and a transient detection light beam with the same intensity, the steady-state detection light beam is received by the steady-state detector 120 and detected to obtain a fluorescence steady-state signal, and the transient detection light beam is received by the transient detector 130 and detected to obtain a fluorescence transient signal.

[0060] Based on the above-described spectroscopy system, the present invention provides a spectroscopy device combined with the spectroscopy system.

[0061] See also Figure 2 The spectral device includes the above-mentioned spectral system, an industrial computer, a servo driver, a main control board, a servo, a stepper motor, an oscilloscope and a high-voltage controller.

[0062] The industrial computer is connected to the servo drive, main control board, laser, spectrometer, oscilloscope and camera. The user can input various parameters on the industrial computer. The industrial computer sends corresponding adjustment instructions to the servo drive, main control board, laser, spectrometer and oscilloscope according to the user input parameters, and at the same time receives data from the camera and displays the picture on the screen of the industrial computer.

[0063] The servo driver is connected to the servo, and sends corresponding control instructions according to the adjustment instructions of the industrial computer to control the displacement change of the servo; the main control board is connected to the stepper motor and the high-voltage controller, and sends corresponding control instructions according to the adjustment instructions of the industrial computer to control the displacement change of the stepper motor and the operation of the PMT detector in the spectrometer.

[0064] The servo includes an attenuator servo, an observation switching servo, and a positioning servo. The attenuator servo is connected to the attenuator 20, and controls the attenuation degree of the attenuator according to the control command of the servo driver; the observation switching servo is connected to the push-pull rod of the observation device 90, and controls the optical path connection between the eyepiece and the beam splitter 80 and the optical path connection switch between the camera and the beam splitter 80 in the observation device 90 according to the control command of the servo driver; the positioning servo is connected to the positioning reflector 43, and drives the rotation and movement of the positioning reflector 43 according to the control command of the servo driver, so that the spot position of the excitation light beam irradiated on the sample surface moves, and controls the center of the spot to be irradiated in the sample target detection area.

[0065] The stepper motor includes a stage coarse adjustment stepper motor and a stage fine adjustment stepper motor. The stage coarse adjustment stepper motor is connected to the stage 60, and drives the stage 60 to move with a relatively coarse step length according to the control instruction of the main control board, and its adjustment range is large but its precision is low; the stage fine adjustment stepper motor is connected to the stage 60, and drives the stage 60 to move with a relatively fine step length according to the control instruction of the main control board, and its adjustment precision is high but its range is small.

[0066] The oscilloscope is connected to the spectrometer and the APD detector respectively, and presents the fluorescence steady-state signal and the transient signal with clear curve changes according to the adjustment instructions of the industrial computer. Specifically, the oscilloscope is connected to the PMT detector of the spectrometer, receives the intensity and wavelength information of the fluorescence signal, and obtains the fluorescence steady-state signal curve; the oscilloscope receives the intensity and time information of the fluorescence signal during attenuation, and obtains the fluorescence transient signal curve.

[0067] The high voltage controller is connected to the PMT detector of the spectrometer and controls the operation of the PMT detector according to the control instructions of the main control board.

[0068] It can be understood that in the present invention, the output command of the industrial computer is mainly used as the transmission of digital signals, the steering gear driver and the main control board are used as the conversion and transmission of digital signals and analog signals, and the steering gear, stepper motor and high-voltage controller are used as the actual response of analog signals. This circuit control setting not only prevents data packet loss when the adjustment command is transmitted in complex devices, but also prevents the control command from being distorted in the response device, thereby improving the operating efficiency of the equipment and the energy utilization rate.

[0069] In addition, the purpose of the servo and the stepper motor is to drive the corresponding device to produce the target displacement to meet the adjustment requirements of the device. It is not limited that the attenuator can only use the servo but not the stepper motor, and it is also not limited that the branch device can only use the stepper motor but not the servo. The adjustment speed and accuracy of the angle displacement using the servo are higher than that of the distance displacement using the stepper motor, but the cost is also higher. The spectral device of the present invention only shows one embodiment with a higher cost-effectiveness. Those skilled in the art can flexibly match the number and position of the servo and stepper motor according to actual needs.

[0070] The following specifically describes the steady-state detection control process of the spectral device in Example 1 of the present invention:

[0071] Select appropriate parameters on the industrial computer according to the properties of the sample. Specifically, when testing the breast, set the initial wavelength of the spectrometer to 380nm, the end wavelength to 600nm, the step to 1nm, and the waiting time to 100ms; set the average number of the oscilloscope to 10 times, the trigger level to -0.5V, the voltage sensitivity to 0.1V, and the time base scale to 0.001s; set the high voltage parameter of the high voltage controller to 2300V.

[0072] The industrial computer sends a steady-state spectrum detection instruction to the spectrometer, and the spectrometer adjusts its own parameters according to the steady-state spectrum detection instruction, detects the fluorescence steady-state signal information, and inputs the detected steady-state information into a channel of the oscilloscope; the industrial computer sends an adjustment instruction to the oscilloscope through the USB interface, and the oscilloscope sets the average number of times, trigger level and other parameters selected by the user according to the adjustment instruction, and receives the steady-state information detected by the spectrometer to make a corresponding steady-state curve.

[0073] The following specifically describes the transient detection control process of the spectral device in Example 1 of the present invention:

[0074] Select appropriate parameters on the industrial computer according to the properties of the sample. Specifically, when testing the breast, set the oscilloscope's average times to 256 times, the trigger level to 0.5V, the voltage sensitivity to 0.002V, and the time base scale to 0.000000005s.

[0075] The industrial computer sends an adjustment instruction to the light source, and the light source turns off the laser output according to the adjustment instruction; the APD detector detects the fluorescence transient signal information, and inputs the detected transient information into another channel of the oscilloscope; the industrial computer sends an adjustment instruction to the oscilloscope through the USB interface, and the oscilloscope sets the average number of times, trigger level and other parameters selected by the user according to the adjustment instruction, and receives the transient information detected by the APD detector to make a corresponding transient curve.

[0076] Since the steady-state and transient-state detection processes do not conflict with each other, and the oscilloscope uses two channels to receive steady-state and transient-state information respectively, the spectrum device of the present invention can simultaneously detect and display steady-state and transient-state curve information.

[0077] In order to obtain the best observation field and improve energy utilization, the spectrum device of the present invention also includes a backlight servo and a backlight lamp, so that the target detection area of ​​the sample is at the center of the observation field before starting to detect the fluorescent signal, that is, the center of the target detection area is on the optical axis of the first objective lens. The backlight servo is connected to a servo driver, and the servo driver controls the backlight servo according to the adjustment instructions of the industrial computer; the backlight servo is connected to the backlight lamp, and the brightness of the backlight lamp is controlled according to the control instructions of the servo driver.

[0078] The following specifically describes the debugging process of the spectral device before formal testing in Example 1 of the present invention:

[0079] The industrial computer issues instructions to the main control board, which controls the stage stepper motor to move in the y direction, so that the stage pops out and the user can place the sample in the center of the stage. Then the main control board controls the stage stepper motor to move in the y direction again, so that the sample can be retracted for testing.

[0080] The industrial computer issues instructions to the servo driver, which controls the backlight servo through the servo driver to turn on the backlight to illuminate the sample target detection area.

[0081] The sample is observed through a camera, which is connected to an industrial computer through a USB interface, and the image of the camera is observed on the industrial computer. Alternatively, the industrial computer issues instructions to the servo driver, and the servo driver controls the observation switch servo to switch to the eyepiece observation state.

[0082] On the main control board, the stage coarse adjustment stepper motor is used to control the stage to coarsely adjust along the x direction, y direction, and z direction respectively; the stage fine adjustment stepper motor is used to control the stage to finely adjust along the x direction, y direction, and z direction respectively; the operation is repeated to focus so that the sample target detection area is in the center of the observation field of view and is clear.

[0083] Control the microscope backlight servo on the servo control panel to turn off the backlight.

[0084] The industrial computer sends instructions through the serial port to enable the laser and turn on the laser.

[0085] The industrial computer sends instructions to the servo driver via the USB-to-serial port connection to adjust the attenuator to the user-specified size, preferably 50%-70%, so as to have the highest possible signal-to-noise ratio without overexposure.

[0086] The industrial computer sends instructions to the servo driver through the USB to serial port connection. The servo driver controls the positioning servo to drive the positioning reflector to rotate, thereby adjusting the position of the laser spot so that it is located in the center of the observation field.

[0087] Example 2

[0088] The only difference between the spectral system in this embodiment 2 and that in embodiment 1 is that the branching device 110 is an optical path switch, and the optical path switch can change the connectivity selection of the optical path. It can be selected to make all the detection beams be input as steady-state detection beams into the steady-state detector 120, or it can be selected to make all the detection beams be input as transient detection beams into the transient detector 130, so as to minimize the loss of fluorescence signal intensity when the sample fluorescence signal is weak, and ensure that the signal intensity is high enough when entering the steady-state detector 120 or the transient detector 130, and the detection signal-to-noise ratio is high.

[0089] The following specifically describes the light beam action process of the spectral system in Example 2 of the present invention:

[0090] The light source 10 emits an initial light beam, which forms an energy-attenuated adjustment light beam after passing through the attenuator 20, and the adjustment light beam forms an expanded light beam after being expanded by the beam expander 30; the expanded light beam is irradiated on the focusing lens 50 after passing through the first reflector 41 and the second reflector 42, and forms an excitation light beam after being focused by the focusing lens 50 and reflected by the positioning reflector 43, and the excitation light beam is irradiated on the sample surface of the stage 60 to excite the sample to form a fluorescent light beam; the fluorescent light beam is focused by the first objective lens 71 to form a microscopic light beam, and the microscopic light beam is reflected by the beam splitter 80 to form an observation light beam, which is reflected by the beam splitter 80 The transmission forms a light beam to be detected; the observation light beam enters the observation device 90 and is directly or indirectly observed to obtain the fluorescence state of the sample; the light beam to be detected is focused after passing through the second objective lens 72 to form an amplified light beam, and the amplified light beam is coupled into the transmission optical fiber 100 through the end face of the transmission optical fiber 100 to form a detection light beam; when the branching device 110 is connected to the steady-state detector 120, the detection light beam is received by the steady-state detector 120 and detected to obtain a fluorescence steady-state signal; when the branching device 110 is connected to the transient detector 130, the detection light beam is received by the transient detector 130 and detected to obtain a fluorescence transient signal.

[0091] Correspondingly, the stepper motor in the spectral device in this embodiment 2 also includes a branch switching stepper motor, which is connected to the branch device 110 to control the switching between the optical path connection between the steady-state detector 120 and the transmission optical fiber 100 and the optical path connection between the transient detector 130 and the transmission optical fiber 100 in the branch device 110.

[0092] The following specifically describes the steady-state detection control process of the spectral device in Example 2 of the present invention:

[0093] The industrial computer sends a steady-state spectrum measurement instruction to the main control board, controls the main control board to control the branch switch device, and configures it into a steady-state spectrum measurement mode. In this mode, the measurement unit is a spectrometer, and the steady-state information detected by its internal PMT detector is input into a channel of the oscilloscope.

[0094] Select appropriate parameters on the industrial computer according to the properties of the sample. Specifically, when testing the breast, set the initial wavelength of the spectrometer to 380nm, the end wavelength to 600nm, the step to 1nm, and the waiting time to 100ms; set the average number of the oscilloscope to 10 times, the trigger level to -0.5V, the voltage sensitivity to 0.1V, and the time base scale to 0.001s; set the high voltage parameter of the high voltage controller to 2300V.

[0095] The industrial computer sends a steady-state spectrum detection instruction to the spectrometer, and the spectrometer adjusts its own parameters according to the steady-state spectrum detection instruction, detects the fluorescence steady-state signal information, and inputs the detected steady-state information into a channel of the oscilloscope; the industrial computer sends an adjustment instruction to the oscilloscope through the USB interface, and the oscilloscope sets the average number of times, trigger level and other parameters selected by the user according to the adjustment instruction, and receives the steady-state information detected by the spectrometer to make a corresponding steady-state curve.

[0096] The following specifically describes the transient detection control process of the spectral device in Example 2 of the present invention:

[0097] The industrial computer sends a transient spectrum measurement instruction to the main control board, controls the main control board to control the branch switching device, and configures it into a transient spectrum measurement mode. In this mode, the measurement unit is an APD detector, and the transient information detected by the APD detector is input into another channel of the oscilloscope.

[0098] Select appropriate parameters on the industrial computer according to the properties of the sample. Specifically, when testing the breast, set the oscilloscope's average times to 256 times, the trigger level to 0.5V, the voltage sensitivity to 0.002V, and the time base scale to 0.000000005s.

[0099] The industrial computer sends an adjustment instruction to the light source, and the light source turns off the laser output according to the adjustment instruction; the APD detector detects the fluorescence transient signal information, and inputs the detected transient information into another channel of the oscilloscope; the industrial computer sends an adjustment instruction to the oscilloscope through the USB interface, and the oscilloscope sets the average number of times, trigger level and other parameters selected by the user according to the adjustment instruction, and receives the steady-state information detected by the APD detector to make a corresponding transient curve.

[0100] Except for the above differences, the rest of the structure of the spectral device in the second embodiment is completely the same as that of the spectral device in the first embodiment, which will not be described again here.

[0101] The spectral equipment of the present invention ensures that the optical path can use cheap and low-cost devices through detailed parameter settings and matching of the spectral system, and enables steady-state detection and transient detection to be implemented in the same optical path structure, greatly improving energy utilization and signal collection efficiency, thereby further reducing system costs; at the same time, the spectral system is adjusted by controlling each servo and stepper motor, so that the composition and molecular structure information of each part of the sample can be fully detected.

[0102] The above-mentioned embodiments only express several implementation methods of the present invention, and the description is relatively specific and detailed, but it cannot be understood as limiting the scope of the invention patent. It should be pointed out that for ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the present invention, and the present invention is also intended to include these modifications and modifications.

Claims

1. A spectral system, characterized in that: include A light source that continuously emits an initial beam of light of a certain wavelength; A stage, carrying a sample that is irradiated by the initial light beam and excites a fluorescent signal, wherein the fluorescent signal forms a fluorescent light beam; A first objective lens amplifies the fluorescent light beam to form a microscopic light beam; a second objective lens, which amplifies the microscopic light beam to form an amplified light beam; A transmission optical fiber couples the amplified light beam into the transmission optical fiber to form a detection light beam; A branching switching device, which divides the detection beam into a steady-state detection beam and a transient detection beam, and controls the output channels of the steady-state detection beam and the transient detection beam; A steady-state detector receives a steady-state detection beam and detects and obtains a steady-state fluorescence signal of the sample; A transient detector receives a transient detection beam and detects a transient fluorescence signal of the sample; Wherein, the first objective lens is a long working distance objective lens, so that the initial light beam can be incident on the sample at a large angle.

2. The spectral system according to claim 1, characterized in that: The branch switching device is an optical path switcher, and the connectivity selection of the optical path can be changed by changing the optical path switch, so that all detection beams are input into the steady-state detector as steady-state detection beams or all detection beams are input into the transient detector as transient detection beams.

3. The spectral system according to claim 1, characterized in that: The branch switching device is a beam splitter, which divides the detection beam into a steady-state detection beam and a transient detection beam with the same intensity at an energy splitting ratio of 50 / 50.

4. The spectral system according to claim 2 or 3, characterized in that: It also includes a beam splitter and an observation device; the beam splitter is arranged between the first objective lens and the second objective lens, and transmits the microscopic light beam with a certain energy ratio to form a light beam to be detected, and reflects the microscopic light beam to form an observation light beam, and the light beam to be detected forms an amplified light beam after passing through the second objective lens, and the observation light beam is received by the observation device; or the beam splitter is arranged between the first objective lens and the second objective lens, and reflects the microscopic light beam with a certain energy ratio to form a light beam to be detected, and transmits the microscopic light beam to form an observation light beam, and the light beam to be detected forms an amplified light beam after passing through the second objective lens, and the observation light beam is received by the observation device; the observation device includes an eyepiece, a camera and a push-pull rod, and the position of the push-pull rod is changed to switch between the eyepiece receiving the observation light beam or the camera receiving the observation light beam.

5. The spectral system according to claim 4, characterized in that: It also includes a positioning reflector that reflects the initial light beam to form an excitation light beam, and the excitation light beam is irradiated onto the sample surface. The positioning reflector is rotatable so that the excitation light beam is irradiated onto the target detection area of ​​the sample.

6. The spectral system according to claim 5, characterized in that: It also includes an attenuator and a beam expander. The attenuator is set in the emitting direction of the initial light beam to attenuate the energy of the initial light beam to form an adjusted light beam; the beam expander expands the adjusted light beam to obtain an extended light beam, expands the beam diameter to reduce the beam energy density, and the extended light beam is reflected by the positioning reflector to form an excitation light beam.

7. A spectroscopy device, characterized in that: It comprises the spectral system as described in any one of claims 1 to 6, an industrial computer, a servo drive, a main control board, multiple servos and multiple stepper motors; the industrial computer is connected to the light source, the steady-state detector, the oscilloscope, the servo drive, the main control board and the observation device, and sends corresponding adjustment instructions to the light source, the steady-state detector, the oscilloscope, the servo drive and the main control board according to known input parameters, and simultaneously receives data from the observation device and displays the picture on the screen of the industrial computer; the servo drive is connected to each servo, and sends corresponding control instructions to control the displacement change of the corresponding servo according to the adjustment instructions of the industrial computer; the main control board is connected to each stepper motor, and sends corresponding control instructions to control the displacement change of the corresponding stepper motor according to the adjustment instructions of the industrial computer.

8. The spectral device according to claim 7, characterized in that: The servo includes an attenuator servo, an observation switching servo and a positioning servo; the attenuator servo is connected to the attenuator, and controls the attenuation degree of the attenuator according to the control command of the servo driver; the observation switching servo is connected to the push-pull rod of the observation device, and controls the optical path connection between the eyepiece and the spectrometer and the optical path connection switch between the camera and the spectrometer in the observation device according to the control command of the servo driver; the positioning servo is connected to the positioning reflector, and drives the rotation and movement of the positioning reflector according to the control command of the servo driver, so that the position of the light spot irradiated by the excitation light beam on the sample surface moves, and controls the center of the light spot to be irradiated in the sample target detection area.

9. The spectral device according to claim 8, characterized in that: The stepper motor includes a stage coarse adjustment stepper motor and a stage fine adjustment stepper motor; the stage coarse adjustment stepper motor is connected to the stage, and drives the stage to move with coarse steps according to the control instructions of the main control board; the stage fine adjustment stepper motor is connected to the stage, and drives the stage to move with fine steps according to the control instructions of the main control board.

10. The spectral device according to claim 9, characterized in that: It also includes an oscilloscope and a high-voltage controller. The oscilloscope is connected to the steady-state detector and the transient detector respectively, and presents the fluorescent steady-state signal and the transient signal in a clear curve change according to the adjustment instructions of the industrial computer; the high-voltage controller is connected to the PMT detector inside the steady-state detector, and controls the operation of the PMT detector according to the control instructions of the main control board.

11. The spectral device according to claim 10, characterized in that: It also includes a backlight servo and a backlight lamp. The backlight servo is connected to a servo driver. The servo driver controls the backlight servo according to the adjustment instructions of the industrial computer. The backlight servo is connected to the backlight lamp and controls the brightness of the backlight lamp according to the control instructions of the servo driver.

12. A method for controlling a spectral device according to any one of claims 7 to 11, characterized in that: According to the properties of the sample, appropriate parameters are selected on the industrial computer, and a steady-state spectrum detection instruction is sent to the steady-state detector. The steady-state detector adjusts its own parameters according to the steady-state spectrum detection instruction, detects the fluorescence steady-state signal information, and inputs the detected steady-state information into a channel of the oscilloscope; the industrial computer sends an adjustment instruction to the oscilloscope, the oscilloscope sets its own parameters according to the adjustment instruction, receives the steady-state information detected by the steady-state spectrometer, and makes a corresponding steady-state curve.

13. The spectral device control method according to claim 12, characterized in that: The industrial computer sends an adjustment instruction to the light source, and the light source turns off the laser output according to the adjustment instruction; the transient detector detects the fluorescence transient signal information, and inputs the detected transient information into another channel of the oscilloscope; The industrial computer sends an adjustment instruction to the oscilloscope, and the oscilloscope sets parameters such as the average number of times and the trigger level selected by the user according to the adjustment instruction, receives the steady-state information detected by the transient detector, and makes a corresponding transient curve.

14. The spectral device control method according to claim 13, characterized in that: Before controlling the spectral device to start detection, improve the accuracy of the spectral device by following the steps below: The industrial computer issues instructions to the main control board, and the main control board controls the stage stepper motor to move, driving the stage to pop out, and after the sample is placed in the center of the stage, the main control board controls the stage stepper motor to move, driving the stage to retract the sample; The industrial computer issues instructions to the servo driver, and the servo driver controls the backlight servo to turn on the backlight to illuminate the sample target detection area; Observe the sample through the observation device, control the coarse adjustment of the stage through the stage coarse adjustment stepper motor on the main control board, and control the fine adjustment of the stage through the stage fine adjustment stepper motor; repeatedly operate to focus so that the target detection area of ​​the sample is clearly in the center of the observation field of view; Control the microscope backlight servo on the servo control panel to turn off the backlight; Sending adjustment instructions to the light source through the industrial computer to generate an initial light beam; Sending an adjustment instruction to the steering gear driver through the industrial computer to adjust the attenuator to a certain degree; The industrial computer sends an adjustment instruction to the steering gear driver, and the steering gear driver controls the positioning steering gear to drive the positioning reflector to rotate, change the position of the laser spot, and make the spot be located in the center of the observation field of view.

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