CN107643253B - Cryopreserved whole blood analyzer based on time domain resolution supercontinuum - Google Patents
Cryopreserved whole blood analyzer based on time domain resolution supercontinuum Download PDFInfo
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Abstract
The invention discloses a cryopreserved whole blood analyzer based on a time domain resolution supercontinuum, which consists of a machine body and internal components assembled in the machine body; the internal components mainly comprise a raspberry laser, a super-continuum spectrum fiber laser, a spectrometer, a sample chamber, a direct current stabilized power supply, a touch screen and auxiliary components. The instrument is used for detecting, identifying, establishing a warehouse, tracing and protecting frozen whole blood by customs. The invention has the advantages that the sample chamber constant temperature and initial calibration method is adopted, so that the uniformity of the test environment and the test repeatability are improved; the time domain sequence spectrum collection is carried out on the test of the frozen whole blood sample, the time domain spectrum characteristic difference of different species extracted by the method is far larger than the spectrum difference of conventional single measurement, and the identification rate of different species is further effectively improved.
Description
Technical Field
The invention relates to a supercontinuum laser spectrum analyzer, in particular to a whole blood analyzer adopting a time domain resolution laser supercontinuum comprehensive spectrum technology, which is suitable for testing, building a library and discriminating a whole blood sample frozen and stored by a human and an animal and belongs to the field of photoelectric imaging.
Background
In the import and export field, strict control measures are mostly adopted for import and export of blood products in various countries. Because the blood components of human and animals contain important biological information such as the genetic characteristics of species, the import and export control of the blood components is very strict, and special instruments and equipment are required for detection.
Due to the particularities of blood products, the requirement for testing equipment is non-contact to avoid denaturation by exposure of blood and harm to the testing personnel. Most of the existing blood detection equipment is based on flow cytometry, needs to sample and dilute blood samples and other operations, has high requirements on detection environment, can only be used for fine detection, and is not suitable for the requirement of customs quick customs clearance high-speed detection. In addition, the import and export of blood products in the world currently follow the regulations of cryopreservation tube storage, customs clearance and transportation, namely products such as whole blood, serum and the like are imported and exported in an environment of minus 40-80 ℃, so that the customs detection is of solid blood products. Thus, biomedical devices on the market are not adequate. At present, the national law rules that the export of blood products of people is forbidden and the export of partial blood products of animals is allowed, because in order to prevent illegal personnel from mixing human blood into animal blood for export, an instrument and a method which can rapidly distinguish the blood products frozen by people from the blood products frozen by animals are needed.
In order to meet the requirement, the invention provides a whole blood analysis method and a corresponding instrument adopting a time domain resolution laser supercontinuum technology, which can carry out rapid detection, library building and identification according to the time domain laser supercontinuum difference of human and animal frozen whole blood, and is convenient for a customs import and export inspection and quarantine department to trace, identify and protect the frozen blood products.
Disclosure of Invention
The invention aims to provide a time domain resolution supercontinuum analyzer which can obtain a time domain laser supercontinuum comprehensive spectrum of human and animal cryopreserved whole blood and is used for detecting, identifying, building a library, tracing and protecting the cryopreserved whole blood by customs.
The invention is realized by the following steps:
the invention provides a cryopreserved whole blood analyzer based on a time domain resolution supercontinuum, which comprises a machine body and an internal component assembled in the machine body; the internal components mainly comprise a raspberry laser, a super-continuum spectrum fiber laser, a spectrometer, a sample chamber, a direct current stabilized power supply, a touch screen and auxiliary components;
the machine body is provided with a USB interface, a keyboard interface, a mouse interface and a mains supply switch besides the internal components; the auxiliary components comprise a USB power supply, a plurality of USB lines, a one-to-two USB line, an HDMI line, a serial port data line, a transmitting optical fiber and a receiving optical fiber;
the supercontinuum fiber laser outputs visible to mid-infrared supercontinuum laser to the transmitting fiber through the fiber output port of the laser, and the other end of the transmitting fiber is connected with the input end of the integrating sphere fiber and used for illuminating the integrating sphere; the inner wall of the integrating sphere is coated with a white matte uniform-light material which can perform uniform light on the multiple diffuse reflection of the entering light; the output end of the integrating sphere optical fiber is connected with a spectrometer optical fiber interface of a spectrometer through a receiving optical fiber, and emergent light of the integrating sphere is introduced into the spectrometer for analysis;
the sample chamber comprises a sample rack, an integrating sphere and a thermostat; the sample rack is made of transparent quartz glass, a freezing tube is arranged in the sample rack, and a frozen whole blood sample is filled in the freezing tube; the thermostat enables the sample chamber to be in a constant room temperature environment, the constant temperature enables the temperature difference between all the tested whole blood samples and the testing environment to be consistent, the temperature rise is kept consistent, and the uniformity of the testing environment is guaranteed;
the direct current stabilized voltage supply converts the input commercial power into direct current and divides the direct current into three paths to output; the first path is used for providing a direct current power supply required by the thermostat; the second circuit is used for supplying power to the super-continuum spectrum optical fiber laser; the third path is used for providing direct-current voltage required by the USB power supply; the USB power supply device is provided with three 5V output power supply ports of a USB interface C, a USB interface B and a USB interface A, wherein the USB port of the spectrometer is respectively connected with the USB interface C and the USB interface of the body through one-to-two USB lines, and when the instrument works, the USB interface C outputs 5V direct current voltage to supply power to the spectrometer through the USB port of the spectrometer; when the instrument does not work, the USB interface has no output, and at the moment, the USB interface of the spectrometer can be connected with an external computer through the USB interface of the machine body to carry out off-line debugging; the USB interface B is connected with the raspberry pi USB interface C through a USB line and used for supplying power to the raspberry pi; the USB interface A is connected with the USB interface of the touch screen through a USB line and used for supplying power to the touch screen;
the raspberry HDMI is connected with the touch screen HDMI of the touch screen through an HDMI line and used for displaying the touch screen; the raspberry pi serial port is connected with the spectrometer serial port through a serial port data line for communication; the raspberry group control port is connected with the thermostat and used for setting the constant temperature and receiving a temperature control signal of the thermostat; the keyboard interface is connected with a raspberry pi USB interface B through a USB line, and an external keyboard is inserted into the keyboard interface to perform keyboard input in the raspberry pi; the mouse interface is connected with a raspberry group USB interface A through a USB line, and an external mouse can be inserted into the mouse interface to perform mouse operation in the raspberry group; the raspberry group is provided with main control software, and a user can perform software operation through an external keyboard and mouse or touch operation on a touch screen to perform man-machine interaction;
the invention provides a frozen whole blood analysis method of time domain resolution supercontinuum comprehensive spectrum, which comprises the following steps:
(1) constant temperature of sample chamber
A user puts the sample rack into the sample chamber, and turns on a mains supply switch of the machine body, and at the moment, the direct-current stabilized power supply generates output, so that the raspberry pi, the supercontinuum fiber laser, the spectrometer, the sample chamber, the touch screen and the thermostat are all electrified to be in a working state; the raspberry is electrified to start the main control software, a user clicks a sample chamber constant temperature button of the main control software, the main control software responds, and a constant temperature value T is set for the thermostat; the thermostat starts to heat or cool, a temperature sensor is arranged in the thermostat, when the thermostat reaches a preset temperature, the thermostat returns a signal to the main control software to finish the constant temperature, and the next step is carried out;
(2) initial scaling
A user clicks an initial calibration button of the operation master control software, the master control software generates a response, and the acquisition integration time A of the spectrometer is set; light entering the integrating sphere by the super-continuum spectrum fiber laser is subjected to multiple diffuse reflections for light homogenization; the sample holder is made of transparent quartz glass, so that light rays are basically not absorbed; the master control software starts the spectrometer to perform spectrum collection on emergent light of the integrating sphere according to the integrating time A, so that a reference spectrum is obtained and stored in a memory inside the raspberry group;
(3) temperature-rising time-domain resolved spectrum acquisition
The user takes out the sample rack, puts the cryopreservation tube filled with the frozen whole blood sample in a low-temperature state into the sample rack, then puts the sample rack into the sample chamber, clicks a test starting button of the main control software, the main control software generates a response, and sets a time interval B of time domain resolution and a total test duration C; after light entering an integrating sphere of the supercontinuum optical fiber laser is subjected to light homogenization through multiple diffuse reflection, the incident light entering a whole blood sample can generate multiple comprehensive effects such as absorption, transmission, reflection, re-emission and the like, then the light is emitted to the wall of the integrating sphere along each direction, part of the light is emitted to the whole blood sample after the diffuse reflection, and the light is introduced into a spectrometer through a receiving optical fiber after the light is repeatedly reflected for multiple times; the master control software starts the spectrometer to perform time domain sequence spectrum collection according to the single integration time A, the time interval B and the total test duration C, and the time domain sequence spectrum collection is stored in a sensor inside the raspberry group; because the whole blood sample and the constant temperature value set by the thermostat have great difference in the testing time C, the whole blood sample is in the rapid heating process, the time-domain sequence spectrum of the whole blood sample also changes along with the time, and the time-domain sequence spectrum characteristics of the whole blood of different species are also different due to the difference of the viscosity, the chromaticity, the absorption and other characteristics in the heating process;
(4) analysis of test results
The user clicks a result analysis button of the operation master control software, the master control software generates a response, each group of time domain sequence spectrum data obtained in the third step is divided by the reference spectrum obtained in the second step to obtain each group of time domain sequence comprehensive transmittance spectrum, and principal component analysis and time domain variation curve fitting are carried out on each group of time domain sequence comprehensive transmittance spectrum to obtain a time domain resolution supercontinuum characteristic vector; and carrying out pattern recognition calculation and cluster analysis on the sample feature vectors of a large number of known species of the basic database to obtain the species of the whole blood sample.
The invention has the advantages that the sample chamber constant temperature and initial calibration method is adopted, so that the uniformity of the test environment and the test repeatability are improved; the method is used for testing the frozen whole blood sample, time domain sequence spectrum collection is carried out, the time domain sequence spectrum also changes along with time due to the temperature difference between the frozen whole blood and the sample chamber, and the time domain sequence spectrum characteristics of the whole blood of different species are different due to the difference of the characteristics such as viscosity, chromaticity, absorption and the like in the temperature rising process, so the time domain spectrum characteristic difference of the different species extracted by the method is far greater than the spectrum difference of the conventional single measurement, and the identification rate of the different species is effectively improved.
Drawings
FIG. 1 is a schematic diagram of the system of the present invention, in which: 1-a fuselage; 2-USB interface C; 3-USB interface B; 4-USB interface A; 5-USB interface of the body; 6-keyboard interface; 7-mouse interface; 8-mains switch; 9-DC voltage-stabilized source; 10-USB power supply; 11-supercontinuum fiber laser; 12-laser fiber output port; 13-emitting fiber; 14-integrating sphere fiber input end; 15-output end of optical fiber of integrating sphere; 16-a sample chamber; 17-sample holder; 18-freezing and storing the tube; 19-whole blood sample; 20-integrating sphere; 21-thermostat; 22-receiving optical fiber; 23-spectrometer fiber interface; 24-spectrometer; 25-spectrometer Serial port; 26-spectrometer USB port; 27-serial port data line; 28-raspberry pi serial port; 29-Raspberry pie control port; 30-raspberry pi USB interface a; 31-Raspberry pie USB interface B; 32-raspberry pi HDMI interface; 33-Raspberry Pi USB interface C; 34-touch screen; 35-HDMI line; 36-touch screen HDMI interface; 37-touch screen USB interface; 38-one-to-two USB lines; 39-USB line; 40-Raspberry pie.
Note: USB, Universal Serial Bus (USB); HDMI, High definition multimedia Interface.
Detailed Description
The specific embodiment of the present invention is shown in fig. 1.
The invention provides a cryopreserved whole blood analyzer based on a time domain resolution supercontinuum, which comprises a machine body 1 and internal components assembled in the machine body 1; the internal components mainly comprise a raspberry pi 40, a super-continuum spectrum fiber laser 11, a spectrometer 24, a sample chamber 16, a direct current stabilized voltage supply 9, a touch screen 34 and auxiliary components;
the main body 1 contains the internal components, and also comprises a main body USB interface 5, a keyboard interface 6, a mouse interface 7 and a mains switch 8; the auxiliary components comprise a USB power supply 10, a plurality of USB lines 39, a one-to-two USB line 38, an HDMI line 35, a serial port data line 27, a transmitting optical fiber 13 and a receiving optical fiber 22;
the supercontinuum fiber laser 11 outputs visible to mid-infrared (400 to 1700 nm in this embodiment) supercontinuum laser to an emission fiber 13 through a laser fiber output port 12, and the other end of the emission fiber 13 is connected with an integrating sphere fiber input end 14 for illuminating an integrating sphere 20; the inner wall of the integrating sphere 20 is coated with a white matte uniform-light material which can perform uniform light on the multiple diffuse reflection of the entering light; the integrating sphere optical fiber output end 15 is connected with a spectrometer optical fiber interface 23 of a spectrometer 24 through a receiving optical fiber 22, and the emergent light of the integrating sphere 20 is introduced into the spectrometer 24 (the detection spectrum range of the spectrometer of this embodiment is 400 to 1700 nanometers) for analysis;
the sample chamber 16 comprises a sample frame 17, an integrating sphere 20 and a thermostat 21; the sample rack 17 is made of transparent quartz glass, the freezing tube 18 is arranged in the sample rack, and the freezing tube 18 is internally provided with a frozen whole blood sample 19; the thermostat 21 enables the sample chamber 16 to be in a constant room temperature environment, the constant temperature enables the temperature difference between all the whole blood samples 19 to be tested and the test environment to be consistent, the temperature rise is kept consistent, and the uniformity of the test environment is guaranteed;
the direct current stabilized voltage power supply 9 converts the input commercial power into direct current and divides the direct current into three paths for output; the first path is used for providing direct current power supply required by the thermostat 21; the second circuit is used for supplying power to the super-continuum spectrum optical fiber laser 11; the third path is used for providing the direct-current voltage required by the USB power supply 10; the USB power supply device 10 is provided with three 5V output power supply ports of a USB interface C2, a USB interface B3 and a USB interface A4, wherein the USB port 26 of the spectrometer is respectively connected with the USB interface C2 and the USB interface 5 of the machine body through a one-to-two USB line 38, and when the device works, the USB interface C2 outputs 5V direct current voltage to supply power to the spectrometer 24 through the USB port 26 of the spectrometer; when the instrument does not work, the USB interface C2 has no output, and at the moment, the spectrometer USB interface 26 can be connected with an external computer through the machine body USB interface 5 for off-line debugging; the USB interface B3 is connected with a raspberry pi USB interface C33 of the raspberry pi 40 through a USB line 39 and used for supplying power to the raspberry pi 40; the USB interface A4 is connected with the touch screen USB interface 37 through a USB line 39 and used for supplying power to the touch screen 34;
the raspberry pi HDMI interface 32 of the raspberry pi 40 is connected to the touch screen HDMI interface 36 of the touch screen 34 through an HDMI line 35, so as to provide display for the touch screen 34; the raspberry pi serial port 28 of the raspberry pi 40 is connected with the spectrometer serial port 25 of the spectrometer 24 through a serial port data line 27 for communication; the raspberry pi control port 29 of the raspberry pi 40 is connected with the thermostat 21 and used for setting the constant temperature and receiving the temperature-controlled temperature signal of the thermostat 21; the keyboard interface 6 is connected with a raspberry pi USB interface B31 through a USB line 39, and an external keyboard is inserted into the keyboard interface 6 to perform keyboard input in a raspberry pi 40; the mouse interface 7 is connected with a raspberry pi USB interface 30 through a USB line 39, and an external mouse can be inserted into the mouse interface 7 to perform mouse operation in a raspberry pi 40; the raspberry pie 40 is provided with main control software, and a user can perform software operation through an external keyboard and mouse or touch operation on the touch screen 34 to perform man-machine interaction;
the invention provides a frozen whole blood analysis method of time domain resolution supercontinuum comprehensive spectrum, which comprises the following steps:
(1) constant temperature of sample chamber
A user puts the sample frame 17 into the sample chamber 16, turns on the commercial power switch 8 of the body 1, and at the moment, the direct-current stabilized power supply 9 generates output, so that the raspberry pi 40, the supercontinuum fiber laser 11, the spectrometer 24, the sample chamber 16, the touch screen 34 and the thermostat 21 are powered on and in a working state; the raspberry pi 40 is powered on to start the main control software, a user clicks a sample chamber constant temperature button of the main control software, the main control software responds, and a constant temperature value T (20 degrees in the embodiment) is set for the thermostat 21; the thermostat 21 starts to heat or cool, a temperature sensor is arranged in the thermostat 21, when the preset temperature T is reached, the thermostat 21 returns a signal to the main control software to finish the constant temperature, and the next step is carried out;
(2) initial scaling
The user clicks an initial calibration button of the main control software, and the main control software generates a response to set the acquisition integration time a (200 ms in this embodiment) of the spectrometer 24; the light entering the integrating sphere 20 from the supercontinuum fiber laser 11 is subjected to multiple diffuse reflections for light homogenization; the sample holder 17 is made of transparent quartz glass, so that light rays are basically not absorbed; the master control software starts the spectrometer 24 to perform spectrum collection on emergent light of the integrating sphere 20 according to the integration time A, so that a reference spectrum is obtained and stored in a memory inside the raspberry group 40;
(3) temperature-rising time-domain resolved spectrum acquisition
The user takes out the sample holder 17, puts the cryopreservation tube 18 containing the frozen whole blood sample 19 in a low temperature state (60 ℃ below zero in this embodiment) into the sample holder 17, then puts the sample holder 17 into the sample chamber 16, and the user clicks a start test button of the main control software, so that the main control software responds, and sets a time interval B (note: step length, 5 seconds in this embodiment) for time domain resolution and a total test duration C (1 minute in this embodiment); after light entering the integrating sphere 20 from the supercontinuum fiber laser 11 is subjected to light homogenization through multiple diffuse reflection, light entering the whole blood sample 19 generates multiple comprehensive effects such as absorption, transmission, reflection, re-emission and the like, then the light irradiates the wall of the integrating sphere 20 along each direction, part of the light is irradiated to the whole blood sample 19 after diffuse reflection, and after multiple times of repetition, the light is introduced into a spectrometer 24 through a receiving optical fiber 22 through an optical fiber output end 15 of the integrating sphere 20; the master control software starts the spectrometer 24 to perform time domain sequence spectrum collection (in this embodiment, 12 groups of comprehensive spectrum data are obtained in total) according to the single integration time A, the time interval B and the total test duration C, and stores the time domain sequence spectrum collection into the sensor in the raspberry pie 40; because the whole blood sample 19 and the constant temperature value set by the thermostat 21 have great difference in the test time C, the whole blood sample 19 is in the rapid heating process, the time-domain sequence spectrum of the whole blood sample will change along with the time, and the time-domain sequence spectrum characteristics of the whole blood of different species will be different due to the difference of the viscosity, the chromaticity, the absorption and other characteristics in the heating process;
(4) analysis of test results
The user clicks a result analysis button of the operation master control software, the master control software generates a response, each group of time domain sequence spectrum data obtained in the third step is divided by the reference spectrum obtained in the second step to obtain each group of time domain sequence comprehensive transmittance spectrum, and principal component analysis and time domain variation curve fitting are carried out on each group of time domain sequence comprehensive transmittance spectrum to obtain a time domain resolution supercontinuum characteristic vector; and after pattern recognition calculation and cluster analysis are carried out on the sample feature vectors of a large number of known species in the basic database, the species of the whole blood sample 19 is obtained.
Claims (1)
1. A cryopreserved whole blood analyzer based on time domain resolution supercontinuum is composed of a machine body (1) and internal components assembled in the machine body (1); the internal components comprise a raspberry (40), a super-continuum spectrum fiber laser (11), a spectrometer (24), a sample chamber (16), a direct current stabilized voltage power supply (9), a touch screen (34) and auxiliary components; the method is characterized in that:
the machine body (1) is provided with a machine body USB interface (5), a keyboard interface (6), a mouse interface (7) and a mains switch (8) besides accommodating the internal components;
the auxiliary components comprise a USB power supply (10), a plurality of USB lines (39), a one-to-two USB line (38), an HDMI line (35), a serial port data line (27), a transmitting optical fiber (13) and a receiving optical fiber (22);
the supercontinuum fiber laser (11) outputs visible to mid-infrared supercontinuum laser to an emission fiber (13) through a laser fiber output port (12), and the other end of the emission fiber (13) is connected with an integrating sphere fiber input end (14) and used for illuminating an integrating sphere (20); the inner wall of the integrating sphere (20) is coated with a white matte uniform-light material, so that the entering light can be subjected to diffuse reflection for multiple times for uniform light; the integrating sphere optical fiber output end (15) is connected with a spectrometer optical fiber interface (23) of a spectrometer (24) through a receiving optical fiber (22), and emergent light of the integrating sphere (20) is introduced into the spectrometer (24) for analysis;
the sample chamber (16) comprises a sample frame (17), an integrating sphere (20) and a thermostat (21); wherein the sample rack (17) is made of transparent quartz glass, the freezing tube (18) is arranged in the sample rack, and the freezing whole blood sample (19) is arranged in the freezing tube (18); the thermostat (21) enables the sample chamber (16) to be in a constant room temperature environment, the constant temperature enables the temperature difference between all the whole blood samples (19) to be tested and the testing environment to be consistent, the temperature rise is kept consistent, and the uniformity of the testing environment is guaranteed;
the direct current stabilized voltage power supply (9) converts the input commercial power into direct current and divides the direct current into three paths for output; the first path is used for providing a direct current power supply required by the thermostat (21); the second path is used for supplying power to the supercontinuum fiber laser (11); the third path is used for providing direct current voltage required by the USB power supply (10); the USB power supply device (10) is provided with three 5V output power supply ports of a USB interface C (2), a USB interface B (3) and a USB interface A (4), wherein the USB port (26) of the spectrometer is respectively connected with the USB interface C (2) and the USB interface (5) of the machine body through a one-to-two USB line (38), and when the instrument works, the USB interface C (2) outputs 5V direct current voltage to supply power to the spectrometer (24) through the USB port (26) of the spectrometer; when the instrument does not work, the USB interface C (2) has no output, and at the moment, the spectrometer USB port (26) can be connected with an external computer through the machine body USB interface (5) for off-line debugging; the USB interface B (3) is connected with a raspberry pi USB interface C (33) of the raspberry pi (40) through a USB line (39) and used for supplying power to the raspberry pi (40); the USB interface A (4) is connected with the touch screen USB interface (37) through a USB wire (39) and used for supplying power to the touch screen (34);
the raspberry pi HDMI (32) of the raspberry pi (40) is connected with the touch screen HDMI (36) of the touch screen (34) through an HDMI line (35) and used for displaying the touch screen (34); a raspberry pi serial port (28) of the raspberry pi (40) is connected with a spectrometer serial port (25) of a spectrometer (24) through a serial port data line (27) for communication; a raspberry pi control port (29) of the raspberry pi (40) is connected with the thermostat (21) and used for setting the constant temperature and receiving a temperature-controlled temperature signal of the thermostat (21); the keyboard interface (6) is connected with a raspberry pi USB interface B (31) through a USB line (39), and an external keyboard can be inserted into the keyboard interface (6) to perform keyboard input in the raspberry pi (40); the mouse interface (7) is connected with a raspberry pie USB interface A (30) through a USB line (39), and an external mouse can be inserted into the mouse interface (7) to perform mouse operation in the raspberry pie (40); the raspberry pie (40) is provided with main control software, and a user can perform software operation through an external keyboard and mouse or touch operation on the touch screen (34) to perform man-machine interaction.
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| JP2005140546A (en) * | 2003-11-04 | 2005-06-02 | Keiichiro Ogawa | Low-temperature diffuse reflection measuring apparatus, sample holder used for the same, and low-temperature diffuse reflection spectrum measuring method |
| CN202502049U (en) * | 2012-02-21 | 2012-10-24 | 中国工程物理研究院流体物理研究所 | Measuring device for refractive indexes of transparent fluids under high pressure |
| CN105628679B (en) * | 2016-03-23 | 2018-06-29 | 中国科学院上海技术物理研究所 | Blood identifier based on infrared Raman Ultraluminescence super continuous spectrums |
| CN105759066B (en) * | 2016-04-26 | 2018-05-22 | 绍兴普施康生物科技有限公司 | Full automatic blood-coagulation detection analysis instrument, method |
| CN207280941U (en) * | 2017-10-13 | 2018-04-27 | 中国科学院上海技术物理研究所 | Whole blood analyser is frozen based on the super continuous integrated spectral of time domain resolution |
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