CN215179635U - Time-resolved test system for anaerobic test of microorganism outer membrane protein - Google Patents

Abstract

The application belongs to the technical field of optical analysis, and particularly relates to a time-resolved test system for anaerobic testing of microorganism outer membrane proteins. A test system for anaerobic test of microorganism outer membrane protein with time resolution comprises a gas displacement device, a sample injection device and a reaction detection device which are connected in sequence, wherein the sample injection device comprises a microorganism anaerobic sample injection mechanism and a reactant anaerobic sample injection mechanism; the reaction detection device comprises a cuvette, an optical fiber spectrometer, an integrating sphere and a light source, wherein incident holes of the light source, the cuvette and the integrating sphere are sequentially arranged along the direction of an optical axis, and the integrating sphere is connected with the optical fiber spectrometer through an optical fiber; the cuvette is provided with a sample inlet and a sample outlet; the output end of the microorganism anaerobic sampling mechanism and the output end of the reactant anaerobic sampling mechanism are both communicated with the sampling port. The method has the advantages of shortening the time interval between the sample introduction and the detection of the microorganisms and the reactants and realizing the effect of hundred millisecond kinetic analysis of the redox state of the outer membrane protein.

Description

Time-resolved test system for anaerobic test of microorganism outer membrane protein

Technical Field

The present application belongs to the technical field of optical analysis. More particularly, it relates to a time-resolved test system for anaerobic testing of microbial outer membrane proteins.

Background

Cytochrome c proteins, which have high redox activity and mediate electron transfer between the inside and the outside of microbial cells, are present on cell membranes of electroactive microorganisms. Their presence is an important reason why microorganisms are able to achieve iron oxide reduction, dye degradation, pollutant conversion. In order to study the redox kinetics of these proteins, researchers need to rapidly capture the changes in their redox states in order to characterize their redox states over time.

The early literature reported that the redox state of outer membrane proteins of microorganisms can be tested by UV-visible spectroscopy, but there are still difficulties in applying the test method to the actual measurement process. Such as: first, the common uv-vis spectrometer cannot directly test the redox state of the microbial outer membrane proteins. This is because the microbial cells can strongly scatter the detection light, making it difficult for the spectrometer to capture the spectrum of the microbial outer membrane proteins, directly leading to a large deviation in the spectroscopic test. Second, the output rate of the spectrum must be within one hundred milliseconds, otherwise the spectrum cannot be recorded quickly. In fact, however, the test requires a spectral scan, which takes time. If an ultraviolet spectrophotometer is used in combination with an integrating sphere (taking a general TU-1901 type spectrophotometer by beijing puolfacture as an example), the scanning speed is set to be "fast", the scanning interval is "1 nm", and the scanning is performed from 600 nm to 300 nm, which takes about 70 seconds, which obviously cannot realize fast spectrum capture. Furthermore, the reaction process for measuring the redox state of the outer membrane protein of microorganisms using spectroscopy is significantly affected by oxygen, which must be measured under anaerobic conditions.

The patent application CN205898667U provides a device for rapidly determining the state of microorganism outer membrane protein, a fiber optic spectrometer and an integrating sphere are matched, and the interference generated by scattered light is eliminated by the integrating sphere, so that the effect of testing the redox state of the microorganism outer membrane protein under a stable condition is realized; in this patent, the fiber optic spectrometer is capable of capturing full spectrum features at speeds on the order of hundreds of milliseconds, thereby achieving the hundreds of milliseconds spectrum capture of the redox state of the microbial outer membrane proteins. In addition, nitrogen flow is used for replacing air in the testing device, so that an oxygen-free condition is created.

With respect to the above-described related art, the inventors have considered that although the spectrometer can record data every hundred milliseconds, accurate detection cannot be achieved for a reaction within 1 second in practice. The reason for this is that for the 1 second reaction process, i.e. the reaction end, a sample injection system of less than 100 milliseconds needs to be provided to effectively shorten the time interval between the samples reaching the cuvette. If the sample introduction time itself exceeds one second, the reaction is finished when the sample reaches the position of the detector, and the dynamic change of the spectrum can not be captured practically. Therefore, monitoring the dynamic change of protein within 1 second requires designing a faster anaerobic sampling device to shorten the time interval between sampling and detection. Thereby realizing dynamic capture of the redox state of the protein on a resolution scale of hundred milliseconds.

SUMMERY OF THE UTILITY MODEL

In order to shorten the time interval between the sample introduction and the detection of microorganisms and reactants and enable a spectrometer to capture the dynamic change of the redox state of the microorganism outer membrane protein from a millisecond scale, the application provides a test system for the anaerobic test of the microorganism outer membrane protein, which is time-resolved. The testing device disclosed by the application can realize hundred millisecond-level sample injection of the outer membrane protein of the microorganism under the anaerobic condition, and complete time-resolved redox state testing of the outer membrane protein of the microorganism, thereby realizing hundred millisecond-level kinetic analysis of the redox state of the outer membrane protein.

The application provides a test system for anaerobic testing of microorganism outer membrane protein with time resolution.

The above purpose of the present application is achieved by the following technical solutions:

a test system for anaerobic test of microorganism outer membrane protein with time resolution comprises a gas displacement device, a sample injection device and a reaction detection device which are sequentially connected, wherein the sample injection device comprises a microorganism anaerobic sample injection mechanism capable of storing a microorganism sample and a reactant anaerobic sample injection mechanism capable of storing a reactant sample; the reaction detection device comprises a cuvette, an optical fiber spectrometer, an integrating sphere and a light source, wherein incident holes of the light source, the cuvette and the integrating sphere are sequentially arranged along the direction of an optical axis, and the integrating sphere is connected with the optical fiber spectrometer through an optical fiber; the cuvette is provided with a sample inlet and a sample outlet; the output end of the microorganism anaerobic sampling mechanism and the output end of the reactant anaerobic sampling mechanism are communicated with the sampling port.

When the time-resolved microorganism outer membrane protein anaerobic test is carried out, because the cuvette is arranged at the incident hole of the integrating sphere, light emitted by the light source can directly pass through the cuvette to enter the integrating sphere and is finally captured by the optical fiber spectrometer. Therefore, when the light source and the fiber optic spectrometer are in working states, the cuvette can be considered to be in a state to be detected at any time. When an experimenter utilizes the microorganism anaerobic sampling mechanism and the reactant anaerobic sampling mechanism to inject the respectively stored sample liquid into the cuvette proportionally and rapidly (namely within hundred milliseconds), the two sample liquids are mixed and start to react, and the optical fiber spectrometer captures and records the dynamic change of the spectrum passing through the cuvette.

By adopting the technical scheme, the time interval between the mixing, reaction and spectral detection of the two sample liquids is shortened to be nearly zero, so that the fiber optic spectrometer can capture the dynamic change of the redox state of the microorganism outer membrane protein from a millisecond scale, and finally the time-resolved microorganism outer membrane protein anaerobic test is successfully realized. The existing devices capable of realizing the hundred millisecond anaerobic sample injection include, but are not limited to, a peristaltic pump, a syringe and the like, and specific parameters of the devices can be set according to the volume of the cuvette and the flow rate of the sample liquid. Such as: the cuvette volume is 1mL, and 1mL of sample is needed in total from the contact of the two sample liquids until the cuvette is filled with the sample liquid, so that when the flow rate of the sample liquid injected into the cuvette exceeds 10mL/s, the sample liquid can be injected into the cuvette within 100ms, and hundred millisecond-level sample injection is realized. Those skilled in the art will appreciate that any device capable of achieving a hundred millisecond anaerobic sample injection should be considered to fall within the scope of the present application.

Of course, it should be understood by those skilled in the art that if the two sample liquids are mixed and redox reaction occurs before entering the cuvette, the anaerobic sampling mechanism can inject the sample mixed liquid into the cuvette within hundreds of milliseconds, and the fiber optic spectrometer can capture the dynamic change of the corresponding spectrum at the initial stage of the redox reaction, thereby completing the anaerobic test. At this time, the parameters of the apparatus may be set according to the volume of the cuvette, the inner diameter of the tube communicating with the cuvette inlet, and the flow rate of the sample liquid. Such as: the length of the tube communicated with the sample inlet of the cuvette is 1cm, the inner diameter of the tube is 0.5cm, the volume of the tube inner space of the tube can be calculated to be 0.20mL according to a cylinder volume formula, the volume of the cuvette is 1mL, namely, the cuvette is filled with the sample liquid from the contact of the two sample liquids, and 1.2mL of samples are needed. Therefore, when the flow rate of the sample liquid in the tube exceeds 12mL/s, the sample liquid can be injected into the cuvette from the tube opening within 100ms, and thus the hundred-millisecond sample injection is realized.

Preferably, the sample outlet and the sample inlet are sequentially arranged along the gravity direction.

It will be understood by those skilled in the art that the arrangement of the sample outlet and the sample inlet along the direction of gravity does not mean that the sample outlet and the sample inlet are strictly distributed along the vector direction of gravity, but means that the included angle between the direction from the sample outlet to the sample inlet and the vector direction of gravity is less than 90 °, and when the cuvette is correctly mounted on the fiber optic spectrometer, the sample inlet should be located below the sample outlet. Therefore, when the sample solution enters the cuvette from the sample inlet, the sample solution is gradually accumulated from the bottom of the cuvette and the liquid level is improved, so that the risks of violent stirring and shaking of the sample solution are reduced, the change of absorbance of the sample solution is reduced, and the accuracy of a detection result is improved.

Preferably, the sample inlet is arranged on the side wall of the cuvette, and the sample outlet is arranged on the top end of the cuvette.

The sample outlet is arranged at the top end of the cuvette and is beneficial to filling the inner cavity of the cuvette with sample solution, so that the difference of the light refractive index between the gas phase and the liquid phase is further reduced, and the accuracy of the spectrum detection is improved.

Preferably, the output end of the microorganism anaerobic sampling mechanism is communicated with a first sampling pipe, the output end of the reactant anaerobic sampling mechanism is communicated with a second sampling pipe, the sampling port is communicated with a mixing pipe, and the first sampling pipe, the second sampling pipe and the mixing pipe form a three-way pipe through a three-way valve.

Through adopting above-mentioned technical scheme, two kinds of sample solutions are premixed in the mixing tube, have better degree of consistency when the solution after the mixture gets into in the cell to the probability that the light refracting index that the density difference that has further reduced the solution leads to appears the deviation, and then improve spectral detection's rate of accuracy.

Preferably, the cuvette is fixedly connected with a sample inlet and a sample outlet, the sample inlet is arranged at the sample inlet, and the mixing tube is elastically sleeved at the sample inlet; the sample outlet nozzle is arranged on the sample outlet, and the sample outlet nozzle is connected with a sample outlet pipe with elasticity.

Through adopting above-mentioned technical scheme, the appearance pipe of advancing and play is managed to be connected with advancing kind mouth and play kind mouth respectively through elasticity, not only makes the appearance pipe of advancing and play kind can with advance kind mouth, play kind mouth firm connection, can carry out timely change to the appearance pipe of advancing and play kind when the circumstances such as jam appears in appearance pipe of advancing and play kind simultaneously to guarantee that the device normally works.

Preferably, the sample injection nozzle comprises a head part and a neck part, the neck part is fixedly connected to the cuvette, the head part is in a circular truncated cone shape, the end with the larger diameter of the head part is fixedly connected with the neck part, the diameter of the end with the larger diameter of the head part is larger than that of the neck part, and a cavity channel for sample circulation is arranged in the head part and the neck part; the structure of the sample outlet nozzle is completely consistent with that of the sample inlet nozzle.

Through adopting above-mentioned technical scheme, the head that is round platform form setting has the effect that the guide inlet tube embolias the introduction nozzle, and the head diameter is greater than the fixed effect that the neck diameter is favorable to reinforcing the inlet tube simultaneously for the inlet tube is difficult for droing from the introduction nozzle.

Preferably, the microorganism anaerobic sample introduction mechanism comprises a first peristaltic pump and a microorganism sample storage container, the first sample introduction pipe is communicated with the microorganism sample storage container, and the first sample introduction pipe is arranged at the working end of the first peristaltic pump; the reactant anaerobic sampling mechanism comprises a second peristaltic pump and a reactant sample storage container, the second sampling pipe is communicated with the reactant sample storage container, and the second sampling pipe is installed at the working end of the second peristaltic pump.

By adopting the technical scheme, the peristaltic pump sucks the sample liquid from each storage container and injects the sample liquid into the cuvette, so that the anaerobic mixing of the sample solution is completed. In addition, the peristaltic pump can reach the rotating speed of more than 500r/min, thereby smoothly realizing the effect of rapid sample introduction within hundred milliseconds.

Preferably, the sample feeding device further comprises a controller, and the first peristaltic pump and the second peristaltic pump are electrically connected to the output end of the controller to control the start and stop of the first peristaltic pump and the second peristaltic pump simultaneously.

The controller can control the first peristaltic pump and the second peristaltic pump to start and stop simultaneously, so that the effect of mixing the microorganism sample solution and the reactant sample solution in a specific proportion simultaneously and finely is achieved.

Optionally, the first sample injection tube and the second sample injection tube are silicone tubes.

The silicone tube has better toughness and can better adapt to the extrusion action of the peristaltic pump.

Optionally, the sampling device further comprises a waste liquid cylinder, and the waste liquid cylinder is communicated with the sample outlet pipe.

The waste liquid after reaction can be pumped into the waste liquid cylinder through the sample outlet pipe, so that the waste liquid is collected, and the environmental friendliness of the experiment is improved.

The utility model discloses following beneficial effect has:

1. this application places microorganism sample solution and reactant sample solution respectively in microorganism anaerobic sampling mechanism and reactant anaerobic sampling mechanism, utilizes two sampling mechanism with two kinds of sample solution direct injection cuvettes for two kinds of sample solutions take place to mix and take place redox reaction in getting into the cuvettes or before getting into the cuvettes, carry out real-time spectrum capture detection to the cuvettes simultaneously. By adopting the method, hundred millisecond-level sample introduction, reaction and spectrum capture of the microbial protein redox state can be effectively realized, so that time-resolved microbial protein redox state hundred millisecond-level kinetic test is smoothly realized on the live bacteria level.

2. The arrangement of the colorimetric ware and the three-way valve which are sealed at the periphery enables sample solution to enter the colorimetric ware gently under the condition of uniform mixing, so that the difference of the internal light refractive index of the colorimetric ware caused by phase difference and solution density difference is reduced, and the accuracy of spectral analysis is improved.

3. The sample inlet nozzle and the sample outlet nozzle which are provided with the head and the neck can improve the connection stability between the mixing tube and the sample outlet tube and the cuvette, thereby providing better protection effect for smooth test.

4. The controller can stably, finely and synchronously control the work of the two peristaltic pumps, thereby ensuring that two sample solutions can be simultaneously injected into the cuvette according to a specific proportion and further ensuring the normal operation of the reaction.

Drawings

FIG. 1 is a schematic diagram showing the structure of a test system for anaerobic assay of time-resolved microbial outer membrane proteins in example 1 of the present application.

Fig. 2 is a schematic structural diagram of a test system in another angle in embodiment 1 of the present application.

FIG. 3 is an exploded view of the cuvette, the mixing tube and the sample discharge tube in example 1 of the present application.

FIG. 4 is a partial cross-sectional view of a test system for anaerobic testing of microbial outer membrane proteins resolved in time according to example 2 of the present application.

Fig. 5 is an enlarged view of the area a in fig. 4.

Fig. 6 is an enlarged view of the region B in fig. 4.

Description of reference numerals: 1. a gas displacement device; 11. a nitrogen gas cylinder; 12. a breather pipe; 2. a sample introduction device; 21. a microorganism anaerobic sampling mechanism; 211. a microbial sample storage cup; 212. a first peristaltic pump; 213. a first syringe; 22. a reactant oxygen-free sample injection mechanism; 221. a reactant sample storage cup; 222. a second peristaltic pump; 223. a second syringe; 23. a three-way pipe; 231. a first sample introduction pipe; 2311. a first ventilation one-way valve; 2312. a first sample introduction one-way valve; 232. a second sample injection pipe; 2321. a second ventilation one-way valve; 2322. a second sample introduction one-way valve; 233. a mixing tube; 234. a three-way valve; 3. a reaction detection device; 31. a cuvette; 311. a sample inlet; 312. a sample outlet; 313. a sample injection nozzle; 3131. a neck portion; 3132. a head portion; 314. a sample outlet nozzle; 3141. a sample outlet pipe; 32. a fiber optic spectrometer; 33. an integrating sphere; 34. a halogen lamp; 4. a waste liquid tank.

Detailed Description

The invention is further described with reference to the drawings and the following examples, which are not intended to limit the invention in any way. Unless otherwise indicated, the reagents, methods and apparatus employed in the present invention are conventional in the art.

Unless otherwise indicated, reagents and materials used in the following examples are commercially available.

Example 1

A test system for time-resolved anaerobic testing of microbial outer membrane proteins, referring to FIG. 1, comprises a gas displacement device 1, a sample introduction device 2, a reaction detection device 3 and a waste liquid cylinder 4.

Referring to fig. 1 and 2, the gas replacement device 1 includes a replacement gas storage container. According to the test requirements, the displacement gas must not contain oxygen and must also be biotoxic. Accordingly, the displacement gas may be selected from most inert gases, such as hydrogen, helium, nitrogen, argon, mixtures of any of the above, and the like. In this embodiment, the replacement gas storage container is a nitrogen gas cylinder 11, the output end of the nitrogen gas cylinder 11 is communicated with a vent pipe 12, and one end of the vent pipe 12 far away from the nitrogen gas cylinder 11 is communicated with the sample injection device 2, so that nitrogen gas in the nitrogen gas cylinder 11 can be sent from the nitrogen gas cylinder 11 to the sample injection device 2 through the vent pipe 12 under high pressure.

Referring to fig. 1 and 2, the sample injection device 2 includes a microorganism anaerobic injection mechanism 21, a reactant anaerobic injection mechanism 22, and a three-way pipe 23. The output end of the microorganism anaerobic sampling mechanism 21 and the output end of the reactant anaerobic sampling mechanism 22 are both communicated with a three-way pipe 23, and the input end of the microorganism anaerobic sampling mechanism 21 and the input end of the reactant anaerobic sampling mechanism 22 are both communicated with the vent pipe 12. Optionally, the vent pipe 12 is arranged in a Y shape, one end of the vent pipe 12 is communicated with the output end of the nitrogen gas cylinder 11, and the other two ends are respectively communicated with the input end of the microorganism anaerobic sampling mechanism 21 and the input end of the reactant anaerobic sampling mechanism 22. However, in this embodiment, there are two nitrogen gas cylinders 11, each nitrogen gas cylinder 11 is communicated with one vent pipe 12, one vent pipe 12 is communicated with the input end of the microorganism anaerobic sampling mechanism 21, and the other vent pipe 12 is communicated with the input end of the reactant anaerobic sampling mechanism 22.

Referring to fig. 2, the microorganism anaerobic sampling mechanism 21 includes a first peristaltic pump 212 and a microorganism sample storage container, and the reactant anaerobic sampling mechanism 22 includes a second peristaltic pump 222 and a reactant sample storage container. It will be understood by those skilled in the art that the microorganism sample storage container and the reagent sample storage container are containers for storing relevant samples, and the containers can be selected from cups, bottles, barrels, tanks, and the like, and specifically, in this embodiment, the microorganism sample storage container is the microorganism sample storage cup 211, and the reagent sample storage container is the reagent sample storage cup 221. The two vent pipes 12 are respectively communicated with the microbial sample storage cup 211 and the reactant sample storage cup 221, the microbial sample storage cup 211 is communicated with the three-way pipe 23 through a first sample inlet pipe 231, in this embodiment, the first sample inlet pipe 231 is a hose used in cooperation with the first peristaltic pump 212, the reactant sample storage cup 221 is also communicated with the three-way pipe 23 through a second sample inlet pipe 232, and in this embodiment, the second sample inlet pipe 232 is a hose used in cooperation with the second peristaltic pump 222.

Referring to fig. 2, in detail, the tee 23 includes a mixing pipe 233 and a three-way valve 234, and the first sample pipe 231, the second sample pipe 232 and the mixing pipe 233 are communicated through the three-way valve 234. The mixing tube 233 leads to the reaction detecting unit 3.

Referring to fig. 2, further, the first sample tube 231 and the second sample tube 232 are silicone tubes with high deformation resistance and extrusion resistance. Further, the first peristaltic pump 212 and the second peristaltic pump 222 are controlled by a controller. In this embodiment, the controller is a single-chip microcomputer having two output ends, one of the output ends of the single-chip microcomputer is electrically connected to the input end of the first peristaltic pump 212, and the other output end of the single-chip microcomputer is electrically connected to the input end of the second peristaltic pump 222. The single chip microcomputer can control the first peristaltic pump 212 and the second peristaltic pump 222 to be started or stopped simultaneously, and in addition, the single chip microcomputer can control the first peristaltic pump 212 and the second peristaltic pump 222 to pump the solution according to the set rotating speed.

Referring to fig. 2, the reaction detecting apparatus 3 includes a cuvette 31, a fiber spectrometer 32, an integrating sphere 33, and a light source, incident holes of the light source, the cuvette 31, and the integrating sphere 33 are sequentially arranged along an optical axis direction, a mixing tube 233 is communicated with the cuvette 31, and the integrating sphere 33 is connected with the fiber spectrometer 32 through an optical fiber. The light source may be a xenon lamp or a halogen lamp 34, and in the present embodiment, the halogen lamp 34 is used as the light source.

Referring to fig. 2 and 3, the cuvette 31 is a small tube made of quartz or glass, the periphery of which is closed, and the inside of which is a cavity (for containing liquid), the cuvette 31 is provided with a sample inlet 311 and a sample outlet 312, the sample inlet 311 is provided on a side wall of the cuvette 31 and is close to one end of the cuvette 31, and the sample outlet 312 is provided at one end of the cuvette 31 far from the sample inlet 311. Further, cell 31 fixedly connected with advances kind mouth 313 and appearance mouth 314, advance kind mouth 313 and the structure of appearance mouth 314 is identical completely, advance kind mouth 313 locate advance kind mouth 311 and with the cavity intercommunication in the cell 31, appearance mouth 314 is located out kind mouth 312 and is communicated with the cavity in the cell 31, the one end cover that the triple valve 234 is kept away from to hybrid tube 233 is located outside advance kind mouth 313, hybrid tube 233 has the elasticity, make hybrid tube 233 can utilize elasticity and advance kind mouth 313 sealing connection. The sample outlet 314 is connected with an elastic sample outlet pipe 3141, and the sample outlet pipe 3141 is sleeved outside the sample outlet 314, so that the sample outlet pipe 3141 can be hermetically connected with the sample outlet 314 by using elasticity.

Referring to fig. 2 and 3, when the cuvette 31 is correctly installed, the cuvette 31 is vertically disposed, and the sample inlet 313 is located below the sample outlet 314, so that the sample outlet 314 and the sample inlet 313 can be sequentially distributed along the gravity direction. Furthermore, the sample injection nozzle 313 includes a neck 3131 and a head 3132, the neck 3131 is a straight-walled tubular shape, one end of the neck 3131 is fixedly connected to the cuvette 31, the other end is fixedly connected to the head 3132, and a cavity for fluid to flow through is formed between the neck 3131 and the head 3132; the head 3132 is in a circular truncated cone shape, the end of the head 3132 with a larger diameter is fixedly connected to the neck 3131, and the end of the head 3132 with a larger diameter is larger than the neck 3131, so that the mixing tube 233 and the head 3132 have a higher connection stability when the mixing tube 233 is tightly fastened to the head 3132 and the neck 3131.

Referring to fig. 1 and 3, the end of the sample outlet pipe 3141 away from the sample outlet nozzle 314 is communicated with the waste liquid tank 4, so that the waste liquid after the reaction is completed can be collected and treated uniformly by the waste liquid tank 4.

The working principle of the embodiment is as follows: the tester opens the valve of the nitrogen gas cylinder 11 in the gas replacement device 1, so that nitrogen gas flows through the vent pipe 12, the microorganism sample storage cup 211 (or the reactant sample storage cup 221), the first sample inlet pipe 231 (or the second sample inlet pipe 232), the mixing pipe 233, the cuvette 31, the sample outlet pipe 3141 and the waste liquid cylinder 4 in sequence under high pressure, thereby completing the gas replacement in the whole test system and enabling the test system to be in an anaerobic environment. Then, the tester closes the valve of the nitrogen gas cylinder 11, the first peristaltic pump 212 and the second peristaltic pump 222 are controlled to be opened by the single chip microcomputer, the first peristaltic pump 212 and the second peristaltic pump 222 simultaneously mix the microorganism sample and the reactant solution in a specific ratio in the mixing tube 233, then inject the mixture into the cuvette 31 through the sample inlet 311, react, and the reacted solution flows out through the sample outlet 312 and enters the waste liquid tank 4. In the reaction process, light emitted by the halogen lamp 34 passes through the solution in the cuvette 31 and then enters the integrating sphere 33 through the incident hole of the integrating sphere 33, and finally the spectral signal is captured by the optical fiber spectrometer 32 and analyzed, so that the time-resolved microorganism outer membrane protein anaerobic test is completed.

Example 2

Referring to fig. 4, the difference from the embodiment 1 is that the microorganism anaerobic sampling mechanism 21 is a first syringe 213, and the microorganism sample solution is stored in the first syringe 213; the reactant anaerobic sampling mechanism 22 is a second injector 223, and the reactant sample solution is stored in the second injector 223.

Referring to fig. 4 and 5, the output end of the first syringe 213 communicates with the side wall of the first sample tube 231, and the end of the first sample tube 231 remote from the three-way valve 234 communicates with the vent tube 12. A first ventilation one-way valve 2311 and a first sample introduction one-way valve 2312 are arranged in the first sample introduction pipe 231, the first ventilation one-way valve 2311 is arranged at the joint of the vent pipe 12 and the first sample introduction pipe 231, and the first ventilation one-way valve 2311 is opened and closed in a one-way mode towards the direction of the first sample introduction pipe 231; the first sample inlet one-way valve 2312 is disposed at a junction of the first injector 213 and the first sample inlet pipe 231, and the first sample inlet one-way valve 2312 is opened and closed in one direction toward the first sample inlet pipe 231. Further, the first injector 213 is disposed near the cuvette 31 such that the sample solution in the first injector 213 can rapidly enter the cuvette 31.

Referring to fig. 4 and 6, the output end of the second syringe 223 is communicated with the side wall of the second sample tube 232, and the end of the second sample tube 232 remote from the three-way valve 234 is communicated with the vent tube 12. A second gas exchange one-way valve 2321 and a second sample introduction one-way valve 2322 are arranged in the second sample introduction pipe 232, the second gas exchange one-way valve 2321 is arranged at the joint of the vent pipe 12 and the second sample introduction pipe 232, and the second gas exchange one-way valve 2321 is opened and closed in one way towards the direction of the second sample introduction pipe 232; the second sample introduction check valve 2322 is disposed at a junction of the second injector 223 and the second sample introduction pipe 232, and the second sample introduction check valve 2322 opens and closes unidirectionally towards the second sample introduction pipe 232. Further, the second injector 223 is disposed adjacent to the cuvette 31 such that the sample solution in the second injector 223 can rapidly enter the cuvette 31.

Referring to fig. 4, the first injector 213 and the second injector 223 are both controlled by a single chip microcomputer to inject the solution at a rate, so that the first injector 213 and the second injector 223 can both inject the corresponding sample solution into the cuvette 31 according to a preset ratio.

The working principle of the embodiment is as follows: the tester opens the valve of the nitrogen gas cylinder 11 in the gas replacement device 1, so that nitrogen gas flows through the vent pipe 12, the first sample inlet pipe 231 (or the second sample inlet pipe 232), the mixing pipe 233, the cuvette 31, the sample outlet pipe 3141 and the waste liquid cylinder 4 in sequence under high pressure, thereby completing the replacement of gas in the whole test system and enabling the test system to be in an oxygen-free environment. Then, the tester closes the valve of the nitrogen gas cylinder 11, the first injector 213 and the second injector 223 are controlled by the single chip microcomputer to inject the microorganism sample and the reactant solution in a specific ratio into the mixing tube 233, the two solutions are mixed in the mixing tube 233 and then injected into the cuvette 31 through the injection port 311 to react, and the reacted solution flows out through the sample outlet 312 and enters the waste liquid tank 4. In the reaction process, light emitted by the halogen lamp 34 passes through the solution in the cuvette 31 and then enters the integrating sphere 33 through the incident hole of the integrating sphere 33, and finally the spectral signal is captured by the optical fiber spectrometer 32 and analyzed, so that the time-resolved microorganism outer membrane protein anaerobic test is completed.

The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be equivalent replacement modes, and all are included in the scope of the present invention.

Claims (10)

1. The utility model provides a test system that time-resolved microorganism adventitia protein anaerobism test was used, includes gas replacement device (1), sample sampling device (2) and reaction detection device (3) that connect gradually, its characterized in that: the sample injection device (2) comprises a microorganism anaerobic sample injection mechanism (21) capable of storing a microorganism sample and a reactant anaerobic sample injection mechanism (22) capable of storing a reactant sample; the reaction detection device (3) comprises a cuvette (31), a fiber optic spectrometer (32), an integrating sphere (33) and a light source, wherein incident holes of the light source, the cuvette (31) and the integrating sphere (33) are sequentially arranged along the direction of an optical axis, and the integrating sphere (33) is connected with the fiber optic spectrometer (32) through an optical fiber; the cuvette (31) is provided with a sample inlet (311) and a sample outlet (312); the output end of the microorganism anaerobic sampling mechanism (21) and the output end of the reactant anaerobic sampling mechanism (22) are communicated with the sampling port (311).

2. The test system of claim 1, wherein: the sample outlet (312) and the sample inlet (311) are sequentially arranged along the gravity direction.

3. The test system of claim 2, wherein: the sample inlet (311) is arranged on the side wall of the cuvette (31), and the sample outlet (312) is arranged on the top end of the cuvette (31).

4. A test system according to any one of claims 1-3, wherein: the output end of the microorganism anaerobic sampling mechanism (21) is communicated with a first sampling pipe (231), the output end of the reactant anaerobic sampling mechanism (22) is communicated with a second sampling pipe (232), the sampling port (311) is communicated with a mixing pipe (233), and the first sampling pipe (231), the second sampling pipe (232) and the mixing pipe (233) form a three-way pipe (23) through a three-way valve (234).

5. The test system of claim 4, wherein: the cuvette (31) is fixedly connected with a sample inlet nozzle (313) and a sample outlet nozzle (314), the sample inlet nozzle (313) is arranged at the sample inlet (311), and the mixing tube (233) is elastically sleeved on the sample inlet nozzle (313); the sample outlet (314) is arranged at the sample outlet (312), and the sample outlet (314) is connected with a sample outlet pipe (3141) with elasticity.

6. The test system of claim 5, wherein: the sample injection nozzle (313) comprises a head part (3132) and a neck part (3131), the neck part (3131) is fixedly connected to the cuvette (31), the head part (3132) is in a circular truncated cone shape, the end of the head part (3132) with the larger diameter is fixedly connected with the neck part (3131), the end of the head part (3132) with the larger diameter is larger than the diameter of the neck part (3131), and channels for sample circulation are arranged in the head part (3132) and the neck part (3131); the structure of the sample outlet nozzle (314) is completely consistent with that of the sample inlet nozzle (313).

7. The test system of claim 4, wherein: the microorganism anaerobic sample feeding mechanism (21) comprises a first peristaltic pump (212) and a microorganism sample storage container, the first sample feeding pipe (231) is communicated with the microorganism sample storage container, and the first sample feeding pipe (231) is installed at the working end of the first peristaltic pump (212); the reactant anaerobic sampling mechanism (22) comprises a second peristaltic pump (222) and a reactant sample storage container, a second sampling pipe (232) is communicated with the reactant sample storage container, and the second sampling pipe (232) is installed at the working end of the second peristaltic pump (222).

8. The test system of claim 7, wherein: the sample feeding device (2) further comprises a controller, and the first peristaltic pump (212) and the second peristaltic pump (222) are electrically connected with the output end of the controller to control the start and stop of the first peristaltic pump (212) and the second peristaltic pump (222) at the same time.

9. The test system of claim 7, wherein: the first sampling tube (231) and the second sampling tube (232) are silicone tubes.

10. The test system of claim 5, wherein: still include waste liquid jar (4), waste liquid jar (4) with go out appearance pipe (3141) intercommunication.

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