1. The utility model provides a micron order unicell light and heat evaluation system based on unsettled thermocouple array which characterized in that includes: the device comprises a laser emission module, a three-dimensional positioning module, a temperature sensing module and a data acquisition and processing module;

the laser emission module is used for outputting laser with adjustable parameters, the three-dimensional positioning module is used for accurately positioning micron-sized laser to irradiate the junction position of a thin-film thermocouple for growth of a single cell on the temperature sensing chip, the temperature sensing module is used for converting temperature change generated after the laser irradiates the single cell into voltage data, the data acquisition and processing module is used for reading and visualizing the acquired voltage data in real time and converting the voltage data into corresponding temperature data according to the Seebeck coefficient of the thin-film thermocouple temperature sensor, so that real-time monitoring and objective quantitative evaluation of the micron-sized laser-single cell photothermal effect are realized;

the laser emission module comprises a laser light source, a control circuit, an optical fiber and an optical fiber connection port; the laser light source is a semiconductor laser coupled and output by optical fibers and used for generating laser; the control circuit is used for selecting and editing required optical parameters, and the optical parameters comprise laser wavelength, intensity, mode, optical pulse width and repetition frequency; the optical fiber connection port is used for transmitting laser energy to an optical fiber, and the optical fiber is used for transmitting laser to a cell to be detected;

the three-dimensional positioning module is an XYZ three-dimensional positioning instrument; the XYZ three-dimensional position indicator is connected with the laser emission module in a mode of clamping an optical fiber;

the temperature sensing module comprises a temperature sensing chip, and the temperature sensing chip consists of a Pd/Cr film thermocouple array; the Pd/Cr film thermocouple array is used for converting temperature change generated after laser irradiation of single cells into weak voltage between Pd and Cr cold end ports; the thermocouple array is connected with the data acquisition and processing module through a DuPont plug wire by externally connecting 4-100 lead wires;

the data acquisition and processing module comprises a multi-channel gate, a nano-volt meter and a LabVIEW control program; the multi-channel gate is used for sequentially opening the passage of each monitored thermocouple so that the nano-volt meter can perform cyclic measurement on the output voltage of each thermocouple, the nano-volt meter is used for reading the voltage data of the monitored thermocouple and transmitting the voltage data to a LabVIEW control program, and the LabVIEW control program is used for controlling the data acquisition range, sequence and interval of the multi-channel gate and displaying the voltage data acquired from the thermocouple array in real time; the multi-channel gate is connected with the temperature sensing module through a DuPont plug wire, and the nanovoltmeter is respectively connected with the multi-channel gate and the LabVIEW control program; the nano-volt meter is connected with the multi-channel gate through a serial port line, and voltage data of the monitored thermocouple are read; the nanovoltmeter transmits voltage data to the LabVIEW control program via GPIB-HS.

2. The suspended thermocouple array-based micro-scale single-cell photothermal evaluation system according to claim 1, wherein the laser light source has a wavelength range including infrared and visible light bands: 450-1,500 nm.

3. The suspended thermocouple array-based micro-scale single-cell photothermal evaluation system according to claim 2, wherein the laser has an intensity range of: the energy density of the unit area of the highest unit time can reach 9549.3J/cm²The maximum power can reach 3W.

4. The suspended thermocouple array-based micro-scale single-cell photothermal evaluation system according to claim 3, wherein the laser light has a mode divided into continuous light and pulsed light.

5. The suspended thermocouple array-based micro-scale single-cell photothermal evaluation system according to claim 4, wherein the pulse width modulation range of the pulse laser is from 10 μ s to infinity, and the pulse repetition frequency range is 0-100 KHz.

6. The suspended thermocouple array-based micro-scale single-cell photothermal evaluation system according to claim 5, wherein the XYZ three-dimensional position finder has a positioning accuracy of 0.03 mm.

7. The suspended thermocouple array-based micro-scale single-cell photothermal evaluation system according to claim 6, wherein the thermocouple array has an array range of: 1X 1 to 10X 10.

8. The suspended thermocouple array-based micron-sized single-cell photothermal evaluation system according to claim 7, wherein the temperature measurement junction region of the Pd-Cr thermocouple array is fabricated on a silicon nitride suspended platform, the cold end is fabricated at the edge of the chip, the temperature measurement accuracy is high 21 ± 0.1 μ V/K, and the thermal noise at room temperature is 20 mK.

9. The suspended thermocouple array-based micro-scale single-cell photothermal evaluation system according to any one of claims 1-8, wherein when the multi-channel gate simultaneously monitors four thermocouple units on the same silicon nitride window, all the thermocouple units can be measured once every 0.3s or so.

Background

The photothermal effect of laser-biological tissue plays an important role not only in medical cutting and rehabilitation therapy, but also in the study of photomodulation of nerve excitation. More and more studies have demonstrated that light can directly cause neuronal excitation under non-transgenic conditions. However, the mechanism by which laser light causes excitation of non-genetically transfected nerve or spiral ganglion cells is not known to date and is now generally believed by the academia to be the result of the action of photoacoustic and photothermal effects, either alone or in combination. The photothermal response of the laser in each frequency range is systematically researched, and the method has very important significance for evaluating the thermal safety of the medical laser and determining the mechanism of photoinduced neural excitation.

At present, most of researches on laser-tissue photothermal effect evaluation are carried out by grading a pain grading scale through subjective feelings of a subject, and are not objective; or the photothermal effect of centimeter-level or millimeter-level laser is evaluated for temperature change, objectively but lacking in accuracy. Considering the accuracy of stimulation, studies on light induced neural excitation often employ laser beams with a diameter of micrometers. However, an effective temperature measurement method and system for the micro-scale laser-tissue photothermal effect evaluation are lacked at present; firstly, the micron-sized laser beam determines the micron-sized requirement on the temperature sensor; secondly, the micrometer-sized laser beam is difficult to locate to the micrometer-sized sensing region with the naked eye and manual operation. Therefore, the application designs a system for objectively and quantitatively evaluating the photothermal effect of the laser-cell interaction on a micrometer scale.

Disclosure of Invention

Aiming at the technical defects, the invention aims to overcome the defects of the existing laser photo-thermal evaluation method in the aspects of objectivity or accuracy, and provides a micron-sized single-cell photo-thermal evaluation system based on a suspended thermocouple array.

In order to solve the technical problems, the invention adopts the following technical scheme:

the utility model provides a micron order unicell light and heat evaluation system based on unsettled thermocouple array which characterized in that includes: the device comprises a laser emission module, a three-dimensional positioning module, a temperature sensing module and a data acquisition and processing module;

the laser emission module is used for outputting laser with adjustable parameters, the three-dimensional positioning module is used for accurately positioning micron-sized laser to irradiate the junction position of a thin-film thermocouple for growth of a single cell on the temperature sensing chip, the temperature sensing module is used for converting temperature change generated after the laser irradiates the single cell into voltage data, the data acquisition and processing module is used for reading and visualizing the acquired voltage data in real time and converting the voltage data into corresponding temperature data according to the Seebeck coefficient of the thin-film thermocouple temperature sensor, so that real-time monitoring and objective quantitative evaluation of the micron-sized laser-single cell photothermal effect are realized;

the laser emission module comprises a laser light source, a control circuit, an optical fiber and an optical fiber connection port; the laser light source is a semiconductor laser coupled and output by optical fibers and used for generating laser; the control circuit is used for selecting and editing required optical parameters, and the optical parameters comprise laser wavelength, intensity, mode, optical pulse width and repetition frequency; the optical fiber connection port is used for transmitting laser energy to an optical fiber, and the optical fiber is used for transmitting laser to a cell to be detected;

the three-dimensional positioning module is an XYZ three-dimensional positioning instrument; the XYZ three-dimensional position indicator is connected with the laser emission module in a mode of clamping an optical fiber;

the temperature sensing module comprises a temperature sensing chip, and the temperature sensing chip consists of a Pd/Cr film thermocouple array; the Pd/Cr film thermocouple array is used for converting temperature change generated after laser irradiation of single cells into weak voltage between Pd and Cr cold end ports; the thermocouple array is connected with the data acquisition and processing module through a DuPont plug wire by externally connecting 4-100 lead wires;

the data acquisition and processing module comprises a multi-channel gate, a nano-volt meter and a LabVIEW control program; the multi-channel gate is used for sequentially opening the passage of each monitored thermocouple so that the nano-volt meter can perform cyclic measurement on the output voltage of each thermocouple, the nano-volt meter is used for reading the voltage data of the monitored thermocouple and transmitting the voltage data to a LabVIEW control program, and the LabVIEW control program is used for controlling the data acquisition range, sequence and interval of the multi-channel gate and displaying the voltage data acquired from the thermocouple array in real time; the multi-channel gate is connected with the temperature sensing module through a DuPont plug wire, and the nanovoltmeter is respectively connected with the multi-channel gate and the LabVIEW control program; the nano-volt meter is connected with the multi-channel gate through a serial port line, and voltage data of the monitored thermocouple are read; the nanovoltmeter transmits voltage data to the LabVIEW control program via GPIB-HS.

Further, the optical wavelength range of the laser light source includes infrared light and visible light bands: 450-1,500 nm.

Further, the intensity range of the laser is as follows: the energy density of the unit area of the highest unit time can reach 9549.3J/cm²The maximum power can reach 3W.

Further, the laser has a mode divided into continuous light and pulsed light.

Further, the pulse width of the pulse laser is adjusted from 10 mus to infinity, and the pulse repetition frequency ranges from 0KHz to 100 KHz.

Furthermore, the positioning precision of the XYZ three-dimensional positioning instrument can reach 0.03 mm.

Further, the array range of the thermocouple array is as follows: 1X 1 to 10X 10.

Furthermore, the temperature measuring junction area of the Pd-Cr thermocouple array is prepared on a silicon nitride suspension platform, the cold end is prepared at the edge of the chip, the temperature measuring precision is high by 21 +/-0.1 mu V/K, and the thermal noise at room temperature is 20 mK.

Furthermore, when the multi-channel gating device simultaneously monitors four thermocouple units on the same silicon nitride window, the measurement can be completed for all the thermocouple units once every 0.3s or so.

The invention has the beneficial effects that: by adopting the technical scheme of the invention, the temperature change caused by the photothermal effect of the micron-sized laser and the single cell can be monitored and evaluated quantitatively in real time, so that a real-time monitoring method and a real-time monitoring system can be provided for the photothermal effect evaluation of the interaction between the micron-sized laser and other biological tissue materials in the future, and basic data can be provided for photothermal effect models of the interaction between the laser and other biological tissue materials.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.

FIG. 1 is a block diagram of a process of a suspended thermocouple array-based micro-scale single-cell photothermal evaluation system according to the present invention;

FIG. 2 is a schematic diagram of a laser precise positioning process of a three-dimensional positioning module of a suspended thermocouple array-based micro-scale single-cell photothermal evaluation system according to the present invention;

FIG. 3 is a schematic diagram of a laser precise positioning process of a three-dimensional positioning module of a suspended thermocouple array-based micro-scale single-cell photothermal evaluation system according to the present invention;

FIG. 4 is a schematic diagram of a laser precise positioning process of a three-dimensional positioning module of a suspended thermocouple array-based micro-scale single-cell photothermal evaluation system according to the present invention;

fig. 5 is a voltage waveform diagram of a single-cell photothermal effect drawn by a suspended thermocouple array-based micro-scale single-cell photothermal evaluation system LabVIEW control program.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

As shown in fig. 1 to 5, a suspended thermocouple array-based micro-scale single-cell photothermal evaluation system includes: the device comprises a laser emission module, a three-dimensional positioning module, a temperature sensing module and a data acquisition and processing module;

the laser emission module is used for outputting laser with adjustable parameters, the three-dimensional positioning module is used for accurately positioning micron-sized laser to irradiate the junction position of a thin-film thermocouple for growth of a single cell on the temperature sensing chip, the temperature sensing module is used for converting temperature change generated after the laser irradiates the single cell into voltage data, the data acquisition and processing module is used for reading and visualizing the acquired voltage data in real time and converting the voltage data into corresponding temperature data according to the Seebeck coefficient of the thin-film thermocouple temperature sensor, so that real-time monitoring and objective quantitative evaluation of the micron-sized laser-single cell photothermal effect are realized;

the laser emission module comprises a laser light source, a control circuit, an optical fiber and an optical fiber connection port; the laser light source is a semiconductor laser coupled and output by optical fibers and used for generating laser; the control circuit is used for selecting and editing required optical parameters, and the optical parameters comprise laser wavelength, intensity, mode, optical pulse width and repetition frequency; the optical fiber connection port is used for transmitting laser energy to an optical fiber, and the optical fiber is used for transmitting laser to a cell to be detected;

the three-dimensional positioning module is an XYZ three-dimensional positioning instrument; the XYZ three-dimensional position indicator is connected with the laser emission module in a mode of clamping an optical fiber;

the temperature sensing module comprises a temperature sensing chip, and the temperature sensing chip consists of a Pd/Cr film thermocouple array; the Pd/Cr film thermocouple array is used for converting temperature change generated after laser irradiation of single cells into weak voltage between Pd and Cr cold end ports; the thermocouple array is connected with the data acquisition and processing module through a DuPont plug wire by externally connecting 4-100 lead wires;

the data acquisition and processing module comprises a multi-channel gate, a nano-volt meter and a LabVIEW control program; the multi-channel gate is used for sequentially opening the passage of each monitored thermocouple so that the nano-volt meter can perform cyclic measurement on the output voltage of each thermocouple, the nano-volt meter is used for reading the voltage data of the monitored thermocouple and transmitting the voltage data to a LabVIEW control program, and the LabVIEW control program is used for controlling the data acquisition range, sequence and interval of the multi-channel gate and displaying the voltage data acquired from the thermocouple array in real time; the multi-channel gate is connected with the temperature sensing module through a DuPont plug wire, and the nanovoltmeter is respectively connected with the multi-channel gate and the LabVIEW control program; the nano-volt meter is connected with the multi-channel gate through a serial port line, and voltage data of the monitored thermocouple are read; the nanovoltmeter transmits voltage data to the LabVIEW control program via GPIB-HS.

Further, the optical wavelength range of the laser light source includes infrared light and visible light bands: 450-1,500 nm.

Further, the intensity range of the laser is as follows: the energy density of the unit area of the highest unit time can reach 9549.3J/cm²The maximum power can reach 3W.

Further, the laser has a mode divided into continuous light and pulsed light.

Furthermore, the pulse width regulation range of the pulse laser is from 10 mus to infinity, the pulse repetition frequency range is 0-100KHz, and the application of the high-precision DAC enables the pulse energy regulation precision to reach 1 muJ/div, so that the pulse energy regulation device can meet different experimental requirements.

Furthermore, the positioning precision of the XYZ three-dimensional positioning instrument can reach 0.03mm, and the laser beam can be accurately positioned and irradiated to the micron-sized thermocouple junction area for single cell growth.

Further, the array range of the thermocouple array is as follows: 1X 1 to 10X 10.

Furthermore, the temperature measuring junction area of the Pd-Cr thermocouple array is prepared on a silicon nitride suspension platform, the cold end is prepared at the edge of the chip, the temperature measuring precision is high by 21 +/-0.1 mu V/K, and the thermal noise at room temperature is 20 mK.

Furthermore, when the multi-channel gating device simultaneously monitors four thermocouple units on the same silicon nitride window, the four thermocouple units can be completely measured once every 0.3s or so, and the monitoring is approximately considered to be simultaneous monitoring.

A method for evaluating micro-scale laser-single cell photothermal effect is shown in FIG. 1, and comprises the following steps: (1) roughly positioning by using a laser emission module; (2) adjusting the three-dimensional positioning module, and accurately positioning the laser beam to a thermosensitive node of the temperature sensing module by using the data acquisition and processing module; (3) and a laser emission module is used for outputting pulse laser to carry out experiments, and a data acquisition and processing module is used for evaluating the single cell photothermal effect.

The micro-scale laser-single cell photothermal effect evaluation experiment of a preferred embodiment is specifically operated as follows:

(1) selecting and editing optical parameters by using a control circuit in a laser emission module, outputting continuous laser with the adjusted optical parameters by a laser source, and roughly irradiating the laser near a Pd/Cr film thermocouple array after the laser is transmitted by an optical fiber connection port and an optical fiber;

(2) an XYZ triaxial positioner of the three-dimensional positioning module is connected with the laser emission module in a mode of clamping an optical fiber, and the scale in the x direction of the XYZ triaxial positioner is adjusted through a LabVIEW control program; as shown in fig. 2, while focusing closely on the voltage waveform graph drawn by the LabVIEW control program, find the scale of x when the voltage value suddenly rises and the positioning instrument is no longer adjusted and the voltage value can be kept at the maximum, as shown in position 3 in fig. 3; keeping the x-direction scale, adjusting the y-direction scale of the positioning instrument, finding the y-direction scale when the voltage value suddenly rises and the positioning instrument is not adjusted any more and the voltage value can be kept at the maximum as shown in fig. 2, and finding the position 6 in fig. 4, wherein the x-direction scale and the y-direction scale of the three-dimensional positioning instrument accurately correspond to the measuring point of the thin-film thermocouple on the temperature sensing module, namely the thermo-sensitive node.

(3) And attaching single cells to the thermosensitive nodes, adjusting a control circuit in the laser emission module, and setting pulse width and pulse frequency to enable the laser light source to output pulse laser, so that the height of the three-dimensional position indicator is reduced for experiments.

(4) The temperature change generated after the laser irradiates the single cell is converted into weak voltage between the Pd and Cr cold end ports by the thermosensitive node, and the thermocouple array is connected with the multi-channel gate through 18 external leads by a DuPont plug wire.

(5) Controlling the data acquisition range, sequence, interval and the like of a multi-channel gate by a LabVIEW control program, sequentially opening the access of each monitored thermocouple by the multi-channel gate, reading the voltage data of the monitored thermocouple by a nanovolt meter connected with a serial port line and the multi-channel gate, and performing circular measurement by matching the monitored thermocouple and the nanovolt meter; when the multi-channel gating device simultaneously monitors four thermocouple units on the same silicon nitride window, the four thermocouple units can be completely measured once every 0.3s or so, and the simultaneous monitoring can be approximately considered.

(6) The voltage data is transmitted to a LabVIEW control program through a GPIB-HS by the nano-volt meter, the collected voltage data is read in real time by the LabVIEW control program, a voltage waveform graph reflecting the photothermal effect of a single cell is drawn, and as shown in figure 5, the voltage data is converted into corresponding temperature data by the data collecting and processing module according to the Seebeck coefficient of the thin-film thermocouple temperature sensor, so that objective and quantitative evaluation of the micron-scale laser-single-cell photothermal effect is realized.

By adopting the technical scheme of the invention, the temperature change caused by the photothermal effect of the micron-sized laser and the single cell can be monitored and evaluated in real time, so that the real-time monitoring method and the real-time monitoring system can be provided for the photothermal effect evaluation of the interaction between the micron-sized laser and other biological tissue materials in the future, and basic data can be provided for the photothermal effect model of the interaction between the laser and other biological tissue materials.

The above-mentioned instructions describe a micro-scale laser single-cell photothermal evaluation system and its specific implementation method in detail, but the present invention is not limited to the protection scope of the present invention, and those skilled in the art should be able to make simple structural changes and other modifications according to the present invention.