CN116223554B - Device and method for detecting dDNP probe molecule multichannel metabolic tracking - Google Patents
Device and method for detecting dDNP probe molecule multichannel metabolic tracking Download PDFInfo
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Abstract
The invention discloses a device and a method for detecting the multi-channel metabolism tracking of a probe molecule of dDNP, which are simple and easy to implement, and a multi-sample parallel transfer high-speed channel is formed by utilizing parts such as a multi-channel switching valve, a fluid sensor, a liquid pump and the like, so that the quantitative high-speed transfer of a hyperpolarized sample transferred by one-time melting can be carried out to a plurality of target detection instruments for multi-channel probe molecule metabolism tracking analysis. The invention can simultaneously study the metabolism conditions of a plurality of different systems, is more convenient and efficient for comparison and analysis among unnecessary systems, has good repeatability for parallel sample study, and has a larger prospect in scientific research or clinical application.
Description
Technical Field
The invention belongs to the technical field of nuclear magnetic resonance, and particularly relates to a device and a method for detecting dDNP probe molecule multichannel metabolic tracking.
Background
Melt dynamic nuclear polarisation (dissolution Dynamic Nuclear Polarization, dnp) is a key technique for improving the detection sensitivity of nuclear magnetic resonance techniques, which can be increased to 4 orders of magnitude or more. The technology comprises the steps of carrying out microwave irradiation on a sample containing free radicals at ultralow temperature (1-4K), transferring high electron saturation polarization degree of the free radicals to probe molecule cores in the sample, rapidly dissolving a solid sample at low temperature by using high-temperature liquid after polarization is completed, transferring the solid sample into an organic system to be observed in NMR/MRI, and observing the metabolic flow direction of target molecules or carrying out metabolic imaging of a target region. The fusion type dynamic nuclear polarization technology has the advantages of ultrahigh sensitivity and in-situ detection, has been tried for real-time metabolic imaging of human tumors, and related preclinical application researches are also ongoing.
In the process of the molten DNP experiment, the preparation of the hyperpolarized molecule takes a long time, and an average experiment takes about 2-3 hours. In order to improve the preparation amount and the preparation efficiency of the hyperpolarized molecular probe, arnaud Comment et al invent a molten DNP polarization device for simultaneously polarizing multiple samples, four samples can be polarized at one time, and then the four samples are sequentially molten and transferred to a target organism for detection, so that the efficiency and the repeatability of an experiment are improved, but the process involves the updating and the improvement of a complex ultralow temperature probe and a molten transfer component, and has high requirements on hardware technology. The specific expression is as follows: 1. the ultralow temperature probe needs to have four independent radio frequency coils to detect the polarization state of the corresponding sample respectively, and has high requirement on the stability of the probe circuit; 2. the increase of the sample causes the increase of the space of the sample area, and for the polarizer, a large-caliber superconducting magnet is needed, so that the construction cost of the instrument is greatly increased; 3. each sample needs to be melted once, the ultra-high temperature solvent is injected into the ultra-low temperature sample area for four times, the melting operation is complex, and once the error occurs, the internal part of the cryostat and the low temperature probe are irreversibly affected.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a device and a method for detecting the dDNP probe molecule multichannel metabolism tracking, the device and the method are simple and easy to implement, a multi-sample parallel transfer high-speed channel is formed by utilizing parts such as a multichannel switching valve, a fluid sensor, a liquid pump and the like, and hyperpolarized sample solution which is transferred from one-time melting can be quantitatively and rapidly transferred to a plurality of target detection instruments for multichannel probe molecule metabolism tracking analysis.
The technical scheme adopted for achieving the purposes of the invention is as follows:
the device for detecting the dDNP probe molecule multichannel metabolic tracking comprises a multichannel switching valve, a quantitative ring, a fluid sensor, a sample inlet, a waste liquid container, a transfer power pipeline, a shunt transfer pipeline and a control module;
the multi-channel switching valve is provided with a first channel and a second channel, the dosing ring is respectively communicated with the first channel and the second channel, and the dosing ring is arranged in the first channel and the second channel;
at least two shunt transfer pipelines are provided, and a sample pipe is arranged at the downstream end of each shunt transfer pipeline;
when the multi-channel valve is switched to the first channel, the sample inlet, the first channel, the quantitative ring, the fluid sensor and the waste liquid container are sequentially communicated to form a complete flow path;
when the multi-channel valve is switched to the second channel, the transfer power pipeline, the second channel, the quantitative ring and each split transfer pipeline are communicated to form a complete flow path, the downstream end of the transfer power pipeline is communicated with the inlet of the second channel, and the upstream end of each split transfer pipeline is communicated with the outlet of the second channel;
the control module is connected with and controls the work of the multi-channel switching valve, the fluid sensor, the transfer power pipeline and each split transfer pipeline.
The transfer power pipeline is sequentially provided with a water container and a first liquid pump along the direction from the upstream end to the downstream end, and the water container is positioned at the upstream end of the transfer power pipeline.
The quantitative ring is formed by winding a pipe into a spiral shape.
The shunt transfer pipeline along the direction of upstream end to downstream end on be equipped with shunt solenoid valve, volumetric flask, second liquid pump, quality detection module and third liquid pump in proper order, be equipped with the level gauge that is used for measuring the liquid level in the volumetric flask on the volumetric flask, the volumetric flask top is provided with the gas vent, control module connects and controls shunt solenoid valve, second liquid pump, quality detection module and third liquid pump's work.
Each shunt transfer pipeline is connected with a recovery pipeline, the recovery pipelines are sequentially provided with a recovery electromagnetic valve and a recovery tank along the direction from the upstream end to the downstream end, the upstream end of each recovery pipeline is connected with the position, located upstream of the corresponding shunt electromagnetic valve, of each shunt transfer pipeline, and the recovery tank is arranged at the downstream end of each recovery pipeline.
The quality detection module comprises a receiving container, a filter head, a heating cable, a first micro cavity, a second micro cavity, a first micro cavity electromagnetic valve, a second micro cavity electromagnetic valve, a syringe and a syringe valve, wherein the filter head is arranged at the inlet of the receiving container, the heating cable is wound on the outer wall of the receiving container, the first micro cavity and the second micro cavity are respectively communicated with the receiving container, the first micro cavity electromagnetic valve is arranged between the first micro cavity and the receiving container, the second micro cavity electromagnetic valve is arranged between the second micro cavity and the receiving container, the first micro cavity is provided with a pH meter for detecting the pH value of the hyperpolarized sample solution in the first micro cavity and a photometer for detecting the concentration of target molecules and free radicals in the hyperpolarized sample solution in the first micro cavity, the second micro cavity is provided with a thermometer for detecting the temperature of the hyperpolarized sample solution in the second micro cavity, and the syringe is communicated with the lower part of the receiving container through the syringe valve.
The quality detection module also comprises a fourth liquid pump which is communicated with the top of the receiving container.
A method for the multichannel metabolic tracer detection of a dnp probe molecule, comprising the steps of:
S 1 switching the multi-channel switching valve to a first channel, and sequentially conducting the sample inlet, the first channel, the quantitative ring, the fluid sensor and the waste liquid container to form a complete flow path, wherein hyperpolarized sample solution enters the quantitative ring through the sample inlet and the first channel;
S 2 when surpassingAfter the quantitative ring is filled with the polarized sample solution, the hyperpolarized sample solution enters the fluid sensor, and when the hyperpolarized sample solution flows through the fluid sensor, the fluid sensor feeds back an electric signal to the control module;
S 3 the control module sends an instruction to the multi-channel switching valve and the transfer power pipeline, the multi-channel switching valve is switched to the second channel, the transfer power pipeline starts to work at the same time, at the moment, the transfer power pipeline, the second channel, the quantitative ring and each split-flow transfer pipeline are conducted to form a complete flow path, liquid in the transfer power pipeline enters the quantitative ring through the second channel, hyperpolarized sample solution in the quantitative ring is pushed to enter each split-flow transfer pipeline, and after passing through the quality detection module of each split-flow transfer pipeline, the hyperpolarized sample solution enters the sample pipe of each split-flow transfer pipeline, and meanwhile the quality detection module carries out quality detection on the hyperpolarized solution;
S 4 and respectively adding the same cell system or different cell systems into at least two sample tubes, and simultaneously carrying out NMR metabolic tracing analysis or MRI metabolic tracing analysis or respectively carrying out NMR metabolic tracing analysis and MRI metabolic tracing analysis on the at least two sample tubes.
Further, the method for diverting the hyperpolarized sample solution by each diverting line is as follows:
P 1 the control module sends instructions to the multi-channel switching valve and the transfer power pipeline and simultaneously sends instructions to the shunt electromagnetic valve, the shunt electromagnetic valve is opened, and the hyperpolarized sample solution is pushed into the volumetric flask from the quantitative ring;
P 2 when the liquid level in the volumetric flask reaches the set liquid level height, the liquid level meter feeds back an electric signal to the control module, the control module sends an instruction, the shunt electromagnetic valve is closed, the second liquid pump is opened, and the hyperpolarized sample solution enters the receiving container;
P 3 after the hyperpolarized sample solution is uniformly mixed with the buffer solution in the receiving container, the control module controls the third liquid pump to be started, the first micro-cavity electromagnetic valve and the second micro-cavity electromagnetic valve, the hyperpolarized sample solution is transferred into the sample tube, and meanwhile, the hyperpolarized sample solution is respectively transferred into the sample tubeAnd the water flows into the first micro cavity and the second micro cavity to detect various indexes.
Further, in step S 1 Before operation, the injector valve of the receiving container is opened, the buffer solution is injected into the receiving container through the injector, and after the injection is completed, the injector valve is closed.
Further, step P 2 In the above, the second liquid pump is turned on and the recovery solenoid valve is turned on, and the redundant hyperpolarized sample solution flows into the recovery tank.
Compared with the prior art, the invention has the advantages that:
1. the invention can improve the transfer speed of the hyperpolarized sample solution and reduce the loss of hyperpolarized state in the process of transferring the hyperpolarized sample solution through the multichannel valve and the programmable liquid pump set.
2. The quality detection module can monitor and control various indexes before the hyperpolarized solution enters the sample tube, and strictly control related experimental conditions.
3. The invention increases the utilization rate of the hyperpolarized sample after melting, improves the experimental efficiency and saves the experimental cost.
4. The invention can simultaneously study the metabolism conditions of a plurality of different systems, is more convenient and efficient for comparison and analysis among unnecessary systems, has good repeatability for parallel sample study, and has a larger prospect in scientific research or clinical application.
5. The invention can control the relative strict experimental condition of the organism system to be observed and carry out parallel experiments.
Drawings
FIG. 1 is a schematic diagram of an apparatus for the multichannel metabolic tracer detection of dDNP probe molecules.
Fig. 2 is a schematic view of a flow path formed by switching the ten-way valve to the first passage.
FIG. 3 is a schematic illustration of a flow path formed by switching a ten-way valve to a second way.
Fig. 4 is a schematic diagram of a first quality detection module.
Wherein, 1-sample inlet; 2-ten channel switching valves; 3-quantitative loop: 4-a fluid sensor; 5-a pool; 6-a first programmable liquid pump; 7-a waste liquid pool; 8-first split transfer line: 81-a first shunt electromagnetic valve, 82-a first volumetric flask, 83-a second programmable liquid pump A, 84-a first quality detection module, 85-a third programmable liquid pump A, 86-a first sample tube, 87-a first liquid level meter, 88-a first air outlet, 89-a first recovery tank, 810-a first recovery electromagnetic valve; 9-second split transfer line: 91-second shunt electromagnetic valve, 92-second volumetric flask, 93-second programmable liquid pump B, 94-second quality detection module, 95-third programmable liquid pump B, 96-second sample tube, 97-second liquid level gauge, 98-second air vent, 99-second recovery tank, 910-second recovery electromagnetic valve; 10-third split transfer line: 101-third shunt electromagnetic valve, 102-third volumetric flask, 103-second programmable liquid pump C, 104-third quality detection module, 105-third programmable liquid pump C, 106-third sample tube, 107-third liquid level gauge, 108-third exhaust port, 109-third recovery tank, 1010-third recovery electromagnetic valve; 11-fourth split transfer line: 111-fourth shunt solenoid valve, 112-fourth volumetric flask, 113-second programmable liquid pump D, 114-fourth quality detection module, 115-third programmable liquid pump D, 116-fourth sample tube, 117-fourth liquid level gauge, 118-fourth vent, 119-fourth recovery tank, 1110-fourth recovery solenoid valve; 12-split manifold; 13-control module.
84-first quality detection module: 8401-first receiving container, 8402-first filter head, 8403-first heating cable, 8404-first micro-cavity A, 8405-second micro-cavity A, 8406-first micro-cavity A electromagnetic valve, 8407-second micro-cavity A electromagnetic valve, 8408-first pH meter, 8409-first photometer, 8410-first infrared thermometer, 8411-fourth programmable liquid pump A, 8412-first injector, 8413-first injector valve.
Detailed Description
The device for detecting the multi-channel metabolism tracing of the dDNP probe molecules according to the invention is detailed below with reference to the accompanying drawings
And (5) fine explanation.
Example 1
The schematic diagram of the device for detecting the dnp probe molecule multichannel metabolic tracking provided in this embodiment is shown in fig. 1, and the device comprises a ten-channel switching valve 2, a quantitative loop 3, a fluid sensor 4, a sample inlet 1, a waste liquid tank 7, a transfer power pipeline, a control module 13, a split manifold 12, a first split transfer pipeline 8, a second split transfer pipeline 9, a third split transfer pipeline 10 and a fourth split transfer pipeline 11.
The ten-way switching valve 2 (AL 10UW, VICI AG International, schenkon, switzerland) has two states, and has a first passage port, a second passage port, a third passage port, a fourth passage port, a fifth passage port, a sixth passage port, a seventh passage port, an eighth passage port, a ninth passage port, and a tenth passage port.
The dosing ring 3 is formed by winding polytetrafluoroethylene tubing into a spiral.
When the ten-way switching valve 2 is in the state one, the first passage port is connected with the second passage port, the third passage port is connected with the fourth passage port, the fifth passage port is connected with the sixth passage port, the seventh passage port is connected with the eighth passage port, the ninth passage port is connected with the tenth passage port, and as shown by a black thick solid line in fig. 2, the second passage port, the first passage port, the dosing ring, the fourth passage port and the third passage port are sequentially conducted to form a first passage.
When the tenth channel switching valve 2 is in the second state, the tenth channel port is connected to the first channel port, the second channel port is connected to the third channel port, the fourth channel port is connected to the fifth channel port, the sixth channel port is connected to the seventh channel port, and the eighth channel port is connected to the ninth channel port, as shown by the black thick solid line in fig. 3, and at this time, the tenth channel port, the first channel port, the dosing ring 3, the fourth channel port and the fifth channel port are sequentially conducted to form the second channel.
The transfer power line is provided with a pool 5 and a first programmable fluid pump 6 (mzr-11508 x 1S,HNP Mikrosysteme GmbH,Schwerin,Germany) in sequence along the direction from the upstream end to the downstream end, the programmable fluid pump being capable of regulating its rotational speed so as to control the flow rate of the transfer fluid, the pool 5 being located at the upstream end of the transfer power line.
The first shunt transfer line 8 is provided with a first recovery line, a first shunt solenoid valve 81, a first volumetric flask 82, a second programmable liquid pump a83, a first quality detection module 84, a third programmable liquid pump a85, and a first sample tube 86 in this order along the direction from the upstream end to the downstream end, the first sample tube 86 being located on the downstream end of the first shunt transfer line 8. The first volumetric flask 82 is provided with a first liquid level meter 87 for measuring the liquid level in the first volumetric flask, the top of the first volumetric flask 82 is provided with a first air outlet 88, and the first air outlet 88 can enable the first volumetric flask 82 to be communicated with the atmosphere. The first recovery pipeline is sequentially provided with a first recovery electromagnetic valve 810 and a first recovery tank 89 along the direction from the upstream end to the downstream end, the upstream end of the first recovery pipeline is connected with the first shunt transfer pipeline 8 at a position upstream of the first shunt electromagnetic valve 81, and the first recovery tank 89 is arranged on the downstream end of the recovery pipeline.
The control module 13 is connected to and controls the operation of the first shunt solenoid valve 81, the first liquid level gauge 87, the second programmable liquid pump a83, the first quality detection module 84, the third programmable liquid pump a85 and the first recovery solenoid valve 810, wherein the first shunt solenoid valve 81 and the first recovery solenoid valve 810 are in a normally closed state.
The second shunt transfer pipeline 9 is sequentially provided with a second recovery pipeline, a second shunt electromagnetic valve 91, a second volumetric flask 92, a second programmable liquid pump B93, a second quality detection module 94, a third programmable liquid pump B95 and a second sample tube 96 along the direction from the upstream end to the downstream end, and the second sample tube 96 is positioned on the downstream end of the second shunt transfer pipeline 9. The second volumetric flask 92 is provided with a second level gauge 97 for measuring the liquid level in the second volumetric flask, the top of the second volumetric flask 92 is provided with a second air outlet 98, and the second air outlet 98 can enable the second volumetric flask 92 to be communicated with the atmosphere. The second recovery line includes a second recovery solenoid valve 910 and a second recovery tank 99 in this order in a direction from an upstream end to a downstream end, the upstream end of the second recovery line is connected to the second shunt solenoid valve 91 at a position upstream of the second shunt solenoid valve 9, and the second recovery tank 99 is disposed at the downstream end of the recovery line.
The control module 13 is connected to and controls the operation of the second shunt solenoid valve 91, the second liquid level gauge 97, the second programmable liquid pump B93, the second quality detection module 94, and the third programmable liquid pump B95, and the second recovery solenoid valve 910, wherein the second shunt solenoid valve 91 and the second recovery solenoid valve 910 are in a normally closed state.
The third shunt transfer pipeline 10 is sequentially provided with a third recovery pipeline, a third shunt electromagnetic valve 101, a third volumetric flask 102, a second programmable liquid pump C103, a third quality detection module 104, a third programmable liquid pump C105 and a third sample tube 106 along the direction from the upstream end to the downstream end, and the third sample tube 106 is positioned on the downstream end of the third shunt transfer pipeline 10. The third volumetric flask 102 is provided with a third liquid level meter 107 for measuring the liquid level in the third volumetric flask, the top of the third volumetric flask 102 is provided with a third air outlet 108, and the third air outlet 108 can enable the third volumetric flask 102 to be communicated with the atmosphere. The third recovery line includes a third recovery solenoid valve 1010 and a third recovery tank 109 in this order in the direction from the upstream end to the downstream end, the upstream end of the third recovery line is connected to the third shunt transfer line 10 at a position upstream of the third shunt solenoid valve 101, and the third recovery tank 109 is disposed at the downstream end of the recovery line.
The control module 13 is connected to and controls the operation of the third shunt solenoid valve 101, the third liquid level gauge 107, the second programmable liquid pump C103, the third quality detection module 104, the third programmable liquid pump C105, and the third recovery solenoid valve 1010, wherein the third shunt solenoid valve 101 and the third recovery solenoid valve 1010 are in a normally closed state.
The fourth shunt transfer line 11 is sequentially provided with a fourth recovery line, a fourth shunt solenoid valve 111, a fourth volumetric flask 112, a second programmable liquid pump D113, a fourth quality detection module 114, a third programmable liquid pump D115, and a fourth sample tube 116 along the direction from the upstream end to the downstream end, and the fourth sample tube 116 is located on the downstream end of the fourth shunt transfer line 11. The fourth volumetric flask 112 is provided with a fourth level gauge 117 for measuring the level of liquid in the fourth volumetric flask, the top of the fourth volumetric flask 112 is provided with a fourth vent 118, and the fourth vent 118 enables the fourth volumetric flask 112 to communicate with the atmosphere. The fourth recovery line includes a fourth recovery solenoid valve 1110 and a fourth recovery tank 119 in this order in the direction from the upstream end to the downstream end, the upstream end of the fourth recovery line is connected to the fourth shunt transfer line 11 at a position upstream of the fourth shunt solenoid valve 111, and the fourth recovery tank 119 is disposed at the downstream end of the recovery line.
The control module 13 is connected to and controls the operation of the fourth shunt solenoid valve 111, the fourth liquid level gauge 117, the fourth programmable liquid pump D113, the fourth quality detection module 114, the fourth programmable liquid pump D115, and the fourth recovery solenoid valve 1110, wherein the fourth shunt solenoid valve 111 and the fourth recovery solenoid valve 1110 are in a normally closed state.
The first mass detection module 84, the second mass detection module 94, the third mass detection module 104 and the fourth mass detection module 114 have the same structure, and for avoiding redundancy, only the structure of the first mass detection module is described herein. As shown in fig. 4, the first mass detection module 84 includes a first receiving container 8401, a first filter head 8402, a first heating cable 8403, a first micro-cavity a8404, a second micro-cavity a8405, a first syringe 8412, a first syringe valve 8413, and a fourth programmable liquid pump a8411. The first filter head 8402 is disposed at an inlet of the first receiving container 8401, and the first heating cable 8403 is wound on an outer wall of the first receiving container 8401, and the heating cable is used for heating and preserving heat of the receiving container. The first syringe 8412 is communicated with the bottom of the receiving container 8401, a first syringe valve 8413 is provided between the first syringe 8412 and the first receiving container 8401, and the first syringe 8412 is used for injecting buffer solution into the receiving container in advance. The first micro-cavity A8404 and the second micro-cavity A8405 are respectively communicated with the lower part of the first receiving container 8401, a first micro-cavity electromagnetic valve A8406 is arranged between the first micro-cavity A8404 and the first receiving container 8401, and a second micro-cavity A electromagnetic valve 8407 is arranged between the second micro-cavity A8405 and the first receiving container 8401. The first microcavity a8404 is provided with a first pH meter 8408 for detecting the pH of the hyperpolarized sample solution within the first microcavity a8404 and a first photometer 8409 for detecting the concentration of target molecules and free radicals in the hyperpolarized sample solution within the first microcavity a 8404. The second minute cavity a8405 is provided with a first infrared thermometer 8410 for detecting the temperature of the hyperpolarized sample solution in the second minute cavity a 8405. The fourth programmable liquid pump a8411 is in communication with the top of the first receiving container 8401, and the fourth programmable liquid pump a8411 is configured to pump excess gas from the hyperpolarized sample solution in the first receiving container 8401 and to expel bubbles therefrom.
It should be noted that: the rotating speeds of the first programmable liquid pump and the second programmable liquid pump are required to be set to be high, the two transfer processes have high requirements on the flow speed of the sample, the transfer efficiency is improved, and the transfer time is shortened. The rotation speed of the third programmable liquid pump needs to be set to a proper rotation speed, the process is to transfer the final quantitative hyperpolarized sample solution into the sample tube, and if the flow speed is too high, the cell solution or the related tissue solution in the sample tube is completely blown away, so the third programmable liquid pump needs to be set to a proper low rotation speed, and the hyperpolarized sample solution is stably transferred into the sample tube.
The control module 13 is connected to and controls the operation of the first pH meter 8408, the first photometer 8409, and the first infrared thermometer 8410.
Manifold 12 communicates with first volumetric flask 82, second volumetric flask 92, third volumetric flask 102, and fourth volumetric flask 112, respectively.
When the ten-channel switching valve 2 is switched to the first channel, the sample inlet 1, the second channel opening, the first channel opening, the dosing ring 3, the fourth channel opening, the third channel opening, the fluid sensor 4 and the waste liquid pool 7 are sequentially conducted to form a complete flow path, as shown by a black thick solid line in fig. 2.
When the ten-way switching valve 2 is switched to the second way, the sump 5, the first programmable liquid pump 6, the tenth passage port, the first passage port, the dosing ring 3, the fourth passage port, the fifth passage port, and the distribution manifold 12 are sequentially conducted to form a complete flow path, as shown by a thick black solid line in fig. 3.
The method of the present invention for the multichannel metabolic tracer assay of dDNP probe molecules is described in detail below in conjunction with the apparatus described above.
Example 2
S 1 First syringe valve 8412 is opened, an appropriate amount of buffer solution (3.3 g TRIS buffer solution) is injected into first receiving container 8401 using first syringe 8412, and the first syringe valve 8412 is closed after the injection is completedA syringe valve 8412; opening a second injector valve, injecting a proper amount of buffer solution into the second receiving container by using the second injector, and closing the second injector valve after the injection is completed; opening a third injector valve, injecting a proper amount of buffer solution into a third receiving container by using a third injector, and closing the third injector valve after the injection is completed; opening a fourth injector valve, injecting a proper amount of buffer solution into a fourth receiving container by using a fourth injector, and closing the fourth injector valve after the injection is completed;
S 2 40mg (1- 13 C) Carrying out low-temperature polarization on a mixed sample of pyruvic acid and 1.3mg TEMPONE free radical, and then using high-temperature solution (the components are 13.3g TRIS buffer solution, 0.6mg EDTA and 2.1mg vitamin C) to melt out the low-temperature polarized sample after the low-temperature polarization is finished, so as to form hyperpolarized sample solution;
S 3 when the hyperpolarized sample solution is melted, the multi-channel switching valve is switched 2 to a first channel, at the moment, the sample inlet 1, the second channel opening, the first channel opening, the quantitative loop 3, the fourth channel opening, the third channel opening, the fluid sensor 4 and the waste liquid pool 7 are sequentially communicated to form a complete flow path, and the hyperpolarized sample solution enters the quantitative loop through the sample inlet 1 and the first channel;
S 4 after the dosing ring 3 is filled with the hyperpolarized sample solution, the hyperpolarized sample solution passes through the fluid sensor 4, when the hyperpolarized sample solution flows through the fluid sensor 4, the fluid sensor 4 feeds back an electric signal to the control module 13, and meanwhile, the hyperpolarized sample solution after flowing through the fluid sensor 4 flows into the waste liquid tank 7;
S 5 after receiving the feedback signal from the fluid sensor 4, the control module 13 sends a command to the ten-channel switching valve 2, the first programmable liquid pump 6, the first shunt solenoid valve 81, the second shunt solenoid valve 91, the third shunt solenoid valve 101 and the fourth solenoid valve 111, and the ten-channel switching valve 2 is switched to the second channel, at this time, the water tank 5, the first programmable liquid pump 6, the tenth channel port, the first channel port, the dosing ring 3, the fourth channel port, the fifth channel port and the shunt manifold 12 are sequentially conducted to form a complete flow path, and at the same time, the first programmable liquid pump 6, the first shunt solenoid valve 81,The second shunt electromagnetic valve 91, the third shunt electromagnetic valve 101 and the fourth shunt electromagnetic valve 111 are opened and work is started, water in the water tank 5 is pumped to push the hyperpolarized sample solution in the quantitative ring 3 to enter the shunt main pipe 12, and then the hyperpolarized sample solution entering the shunt main pipe 12 respectively enters the first volumetric flask 82, the second volumetric flask 92, the third volumetric flask 102 and the fourth volumetric flask 112;
S 6 when the liquid levels in the first volumetric flask 82, the second volumetric flask 92, the third volumetric flask 102 and the fourth volumetric flask 112 reach the set liquid level height, the first liquid level meter 87, the second liquid level meter 97, the third liquid level meter 107 and the fourth liquid level meter 117 feed back electric signals to the control module 13, and the control module 13 controls the first shunt electromagnetic valve 81, the second shunt electromagnetic valve 91, the third shunt electromagnetic valve 101 and the fourth shunt electromagnetic valve 111 to be closed and simultaneously controls the second programmable liquid pump A83, the second programmable liquid pump B93, the second programmable liquid pump C103, the second programmable liquid pump D113, the first recovery electromagnetic valve 810, the second recovery electromagnetic valve 910, the third recovery electromagnetic valve 1010 and the fourth recovery electromagnetic valve 1110 to be opened; then the hyperpolarized sample solution in the first volumetric flask 82 is filtered by the first filter head 8402 and then transferred to the first receiving container 8401 at a high speed, the hyperpolarized sample solution in the second volumetric flask 92 is filtered by the second filter head and then transferred to the second receiving container at a high speed, the hyperpolarized sample solution in the third volumetric flask 102 is filtered by the third filter head and then transferred to the third receiving container at a high speed, the hyperpolarized sample solution in the fourth volumetric flask 112 is filtered by the fourth filter head and then transferred to the fourth receiving container, and meanwhile, the redundant hyperpolarized sample solution flows into the first recovery tank, the second recovery tank, the third recovery tank and the fourth recovery tank respectively;
S 7 after the hyperpolarized sample solution is uniformly mixed with the buffer solution of 8401 in the first receiving container, the control module controls the third programmable liquid pump A85, the first micro-cavity A electromagnetic valve 8406, the second micro-cavity A electromagnetic valve 8407, the first pH meter 8408, the first photometer 8409 and the first infrared thermometer 8410 to be opened, and the hyperpolarized sample solution in the first receiving container 8401 is transferred into the first sample tube at a uniform and proper speed86; meanwhile, the hyperpolarized sample solution in the first receiving container 8401 enters the first micro cavity a8404 and the second micro cavity a8405 respectively, the first pH meter 8408 detects the pH of the hyperpolarized sample solution in the first micro cavity a8404, the first photometer 8409 detects the concentration of target molecules and free radicals in the hyperpolarized sample solution in the first micro cavity a8404, and the first infrared thermometer 8410 detects the temperature of the hyperpolarized sample solution in the second micro cavity a 8405;
the working methods of the second mass detection module, the third mass detection module and the fourth mass detection module are the same as those of the first mass detection module, and finally the hyperpolarized solution entering from the sample inlet is simultaneously split and transferred into the first sample tube 86, the second sample tube 96, the third sample tube 106 and the fourth sample tube 116;
S 8 when the first sample tube 86, the second sample tube 96, the third sample tube 106, and the fourth sample tube 116 are tested by the same instrument (e.g., a nuclear magnetic resonance spectrometer or a nuclear magnetic resonance imager, which is used for performing cell perfusion metabolism NMR metabolic trace analysis, and a nuclear magnetic resonance imager is used for performing MRI metabolic trace analysis):
by using localized nuclear magnetic pulse sequences, the imaging area is divided into a series of matrix areas which are uniformly distributed, samples in each matrix area can be independently sampled, four sample tubes are correspondingly distributed in the four independent matrix areas, and the corresponding results are only displayed in the corresponding matrix, so that NMR/MRI data in the four sample tubes can be obtained.
In practice, the same cell system or different cell systems can be added to each of the four sample tubes. When the injection is different cell systems, observing the metabolism condition of the pyruvic acid in the different cell systems, and carrying out NMR or MRI metabolism tracing analysis; when the injection is the same cell system, the NMR or MRI metabolic tracing analysis of the pyruvic acid in the same cell system can be observed, and the requirement of parallel repeated experiments in biological research can be met.
When four sample tubes are respectively positioned in the nuclear magnetic resonance spectrometer and the nuclear magnetic resonance imager for experiments, different analyses can be performed on the same time dimension, so that the diversity of the experiments is increased and the accuracy of the experiments is improved.
S 9 After the experiment is completed, a proper amount of cleaning solvent ethanol can be added into the water tank 5, the first programmable liquid pump 6, the first shunt electromagnetic valve 81, the second shunt electromagnetic valve 91, the third shunt electromagnetic valve 101 and the fourth shunt electromagnetic valve 111 are started, so that the ethanol can be cleaned through the main pipelines of the first shunt transfer pipeline, the second shunt transfer pipeline, the third transfer pipeline and the fourth transfer pipeline, no residual experimental sample solution exists on the transfer path, and after the cleaning is completed, the residual liquid on the transfer path is completely transferred to the outside of the device by using high-pressure nitrogen.
Claims (11)
1. A device for the multichannel metabolic tracer detection of a dnp probe molecule, characterized in that: the device comprises a multichannel switching valve, a quantitative ring, a fluid sensor, a sample inlet, a waste liquid container, a transfer power pipeline, a shunt transfer pipeline and a control module;
the multi-channel switching valve is provided with a first passage and a second passage, the quantitative ring is connected with the multi-channel switching valve, the quantitative ring is respectively communicated with the first passage and the second passage, and the quantitative ring is arranged in the first passage and the second passage;
at least two shunt transfer pipelines are provided, and a sample pipe is arranged at the downstream end of each shunt transfer pipeline;
when the multi-channel valve is switched to the first channel, the sample inlet, the first channel, the quantitative ring, the fluid sensor and the waste liquid container are sequentially communicated to form a complete flow path;
when the multi-channel valve is switched to the second channel, the transfer power pipeline, the second channel, the quantitative ring and each split transfer pipeline are communicated to form a complete flow path, the downstream end of the transfer power pipeline is communicated with the inlet of the second channel, and the upstream end of each split transfer pipeline is communicated with the outlet of the second channel;
the control module is connected with and controls the work of the multichannel switching valve, the fluid sensor, the transfer power pipeline and each shunt transfer pipeline;
after the multichannel valve is switched to the first passage, when the hyperpolarized sample solution flows through the fluid sensor, the fluid sensor feeds back an electric signal to the control module, and the control module sends an instruction to the multichannel switching valve and the transfer power pipeline to switch the multichannel switching valve to the second passage.
2. The device for the multichannel metabolic tracer detection of dnp probe molecules according to claim 1, wherein: the transfer power pipeline is sequentially provided with a water container and a first liquid pump along the direction from the upstream end to the downstream end, and the water container is positioned at the upstream end of the transfer power pipeline.
3. The device for the multichannel metabolic tracer detection of dnp probe molecules according to claim 1, wherein: the quantitative ring is formed by winding a pipe into a spiral shape.
4. The device for the multichannel metabolic tracer detection of dnp probe molecules according to claim 1, wherein: the shunt transfer pipeline along the direction of upstream end to downstream end on be equipped with shunt solenoid valve, volumetric flask, second liquid pump, quality detection module and third liquid pump in proper order, be equipped with the level gauge that is used for measuring the liquid level in the volumetric flask on the volumetric flask, the volumetric flask top is provided with the gas vent, control module connects and controls shunt solenoid valve, second liquid pump, quality detection module and third liquid pump's work.
5. The device for multi-channel metabolic tracer detection of dnp probe molecules according to claim 4, wherein: each shunt transfer pipeline is connected with a recovery pipeline, the recovery pipelines are sequentially provided with a recovery electromagnetic valve and a recovery tank along the direction from the upstream end to the downstream end, the upstream end of each recovery pipeline is connected with the position, located upstream of the corresponding shunt electromagnetic valve, of each shunt transfer pipeline, and the recovery tank is arranged at the downstream end of each recovery pipeline.
6. The device for multi-channel metabolic tracer detection of dnp probe molecules according to claim 5, wherein: the quality detection module comprises a receiving container, a filter head, a heating cable, a first micro cavity, a second micro cavity, a first micro cavity electromagnetic valve, a second micro cavity electromagnetic valve, a syringe and a syringe valve, wherein the filter head is arranged at the inlet of the receiving container, the heating cable is wound on the outer wall of the receiving container, the first micro cavity and the second micro cavity are respectively communicated with the receiving container, the first micro cavity electromagnetic valve is arranged between the first micro cavity and the receiving container, the second micro cavity electromagnetic valve is arranged between the second micro cavity and the receiving container, the first micro cavity is provided with a pH meter for detecting the pH value of the hyperpolarized sample solution in the first micro cavity and a photometer for detecting the concentration of target molecules and free radicals in the hyperpolarized sample solution in the first micro cavity, the second micro cavity is provided with a thermometer for detecting the temperature of the hyperpolarized sample solution in the second micro cavity, and the syringe is communicated with the lower part of the receiving container through the syringe valve.
7. The apparatus for the multichannel metabolic tracer assay of the dDNP probe molecules according to claim 6, wherein: the quality detection module also comprises a fourth liquid pump which is communicated with the top of the receiving container.
8. A method for the multichannel metabolic tracer detection of a dnp probe molecule, characterized by the steps of:
S 1 switching the multi-channel switching valve to a first channel, and sequentially conducting the sample inlet, the first channel, the quantitative ring, the fluid sensor and the waste liquid container to form a complete flow path, wherein hyperpolarized sample solution enters the quantitative ring through the sample inlet and the first channel;
S 2 after the quantitative ring is filled with the hyperpolarized sample solution, the hyperpolarized sample solution enters the fluid sensor, and when the hyperpolarized sample solution flows through the fluid sensor, the fluid sensor feeds back an electric signal to the control module;
S 3 the control module sends instructions to the multi-channel switching valve and the transfer power pipeline to switch the multi-channel switching valve to the second oneThe path is used for simultaneously transferring the power pipelines, at the moment, the power pipelines, the second path, the quantitative ring and each shunt transfer pipeline are communicated to form a complete flow path, liquid in the power pipelines enters the quantitative ring through the second path, hyperpolarized sample solution in the quantitative ring is pushed to enter each shunt transfer pipeline, and after passing through the mass detection module of each shunt transfer pipeline, the liquid enters the sample pipe of each shunt transfer pipeline, and meanwhile, the mass detection module carries out mass detection on the hyperpolarized solution;
S 4 and respectively adding the same cell system or different cell systems into at least two sample tubes, and simultaneously carrying out NMR metabolic tracing analysis or MRI metabolic tracing analysis or respectively carrying out NMR metabolic tracing analysis and MRI metabolic tracing analysis on the at least two sample tubes.
9. The method for multi-channel metabolic tracer detection of dnp probe molecules according to claim 8, wherein:
in step S 1 Before operation, the injector valve of the receiving container is opened, the buffer solution is injected into the receiving container through the injector, and after the injection is completed, the injector valve is closed.
10. The method for the multichannel metabolic tracer detection of dnp probe molecules according to claim 8, wherein the method for diverting the hyperpolarized sample solution per diverting line is as follows:
P 1 the control module sends instructions to the multi-channel switching valve and the transfer power pipeline and simultaneously sends instructions to the shunt electromagnetic valve, the shunt electromagnetic valve is opened, and the hyperpolarized sample solution is pushed into the volumetric flask from the quantitative ring;
P 2 when the liquid level in the volumetric flask reaches the set liquid level height, the liquid level meter feeds back an electric signal to the control module, the control module sends an instruction, the shunt electromagnetic valve is closed, the second liquid pump is opened, and the hyperpolarized sample solution enters the receiving container;
P 3 after the hyperpolarized sample solution is mixed with the buffer solution in the receiving containerThe control module controls the third liquid pump to be started, the first micro-cavity electromagnetic valve and the second micro-cavity electromagnetic valve, the hyperpolarized sample solution is transferred into the sample tube, and meanwhile, the hyperpolarized sample solution flows into the first micro-cavity and the second micro-cavity respectively, so that various indexes are detected.
11. The method for multi-channel metabolic tracer detection of dnp probe molecules according to claim 10, wherein: step P 2 In the above, the second liquid pump is turned on and the recovery solenoid valve is turned on, and the redundant hyperpolarized sample solution flows into the recovery tank.
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