CN216082456U - Fluorescence labeling rate detecting system - Google Patents
Fluorescence labeling rate detecting system Download PDFInfo
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- CN216082456U CN216082456U CN202122661112.XU CN202122661112U CN216082456U CN 216082456 U CN216082456 U CN 216082456U CN 202122661112 U CN202122661112 U CN 202122661112U CN 216082456 U CN216082456 U CN 216082456U
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Abstract
The application discloses a fluorescence labeling rate detection system, which relates to the technical field of medicine embolism and comprises a sample tank, a filtering device, a fixed container, a diluent tank and a fluorescence detector; the output end of the sample tank is connected with the input end of the filtering device through a first conveyor; the first conveyor is used for conveying the sample liquid; the output end of the filtering device for intercepting the fluorescent marker in the sample liquid is connected with the first input end of the fixed container; the output end of the diluent tank is connected with the second input end of the fixed container through a second conveyor, and the second conveyor is used for conveying diluent; the output end of the fixed container is connected with the input end of the fluorescence detector through a third conveyor; the third conveyor is used for conveying the diluted solution to be measured in the fixed container. The detection system that this application this a quantitative sample, filtration, dilution, detection flow's collocation design formed, not only easy operation is convenient, has also improved the detection precision and has avoided the sample extravagant.
Description
Technical Field
The application relates to the technical field of drug embolization, in particular to a fluorescence labeling rate detection system.
Background
In order to observe the metabolism of the sample in human body, a fluorescent marker is often marked on the sample, and the fluorescence marking rate is an index for measuring the effect of the fluorescent marker.
At present, when the index is measured, the fluorescence detection is often carried out by manually sampling for many times, the operation is complex, errors easily occur, and the manual sampling can not be accurately quantified, so that the sample waste is easily caused.
SUMMERY OF THE UTILITY MODEL
In view of the above, in order to solve the above technical problems, the present application aims to provide a fluorescence labeling rate detection system, which has the following specific technical solutions:
a fluorescence labeling rate detection system, comprising:
the sample tank is used for providing a sample liquid, and a fluorescent marker and a fluorescent monomer for fluorescent labeling are dispersed in the sample liquid;
the diluent tank is used for providing a diluent;
the filter device is used for trapping the fluorescent marker, and the input end of the filter device is connected with the output end of the sample tank through a first conveyor;
a first input end of the fixed container is connected with the output end of the filtering device, and a second input end of the fixed container is connected with the output end of the diluent tank through a second conveyor;
and the input end of the fluorescence detector is connected with the output end of the fixed container through a third conveyor.
In some embodiments, the sample tank is a reaction kettle; and/or
The fluorescent marker is temperature-sensitive nanogel marked with the fluorescent monomer, and the temperature-sensitive nanogel is N-isopropyl acrylamide hydrogel.
In some embodiments, the first conveyor is a peristaltic pump; and/or
The second conveyor is a peristaltic pump; and/or
The third conveyor is a peristaltic pump.
In some embodiments, the fluorescence labeling rate detection system further comprises: a controller;
the controller is electrically connected to the first conveyor, the second conveyor, the third conveyor, and the fluorescence detector.
In some embodiments, the sample tank, the first transporter, the filtration device, and the fixed container are connected in sequence by a transfer pipe; and/or
The diluent tank, the second conveyor and the fixed container are sequentially connected through a conveying pipeline; and/or
The fixed container, the third conveyor and the fluorescence detector are sequentially connected through a conveying pipeline.
In some embodiments, a first control valve is further arranged on the conveying pipeline between the filtering device and the constant volume device; and/or
And a second control valve is also arranged on the conveying pipeline between the second conveyor and the constant volume device.
In some embodiments, the filtration device has filtration pores with adjustable pore size; and/or
The filtering device comprises at least two filters, and the filter pores of the filters are different in pore size.
In some embodiments, the diluent is ultrapure water, PBS buffer, physiological saline, animal serum, or animal whole blood.
In some embodiments, the sample tank is further connected to an agitator provided with an agitating portion for agitating the sample liquid.
In some embodiments, the fluorescence labeling rate detection system further comprises: a waste liquid tank;
the waste liquid tank is connected with the output end of the fluorescence detector.
According to the technical scheme, in the fluorescence labeling rate detection system designed by the application, when the fluorescence labeling rate detection system is applied specifically, sample liquid in a sample tank can be quantitatively conveyed to a fixed container through a first conveyor, and a fluorescence label is intercepted through a filtering device during conveying, so that a solution to be detected conveyed to the fixed container is mainly a fluorescence monomer which is a raw material for forming the fluorescence label and is used for fluorescence labeling; then, quantitatively conveying the dilution liquid in the dilution liquid tank to a constant volume device through a second conveyor to dilute the solution to be detected, so that the concentration of the fluorescent monomer subsequently conveyed to the fluorescence detector meets the requirement of the detection limit of the fluorescence detector, then conveying the diluted solution to be detected to the fluorescence detector through a third conveyor to detect, and converting the detected fluorescence intensity to obtain the fluorescence labeling rate of the fluorescence labeling reaction.
Compared with the prior art, the fluorescent marker detection system designed by the matching of the quantitative sampling, filtering, diluting and detecting processes has the following beneficial effects:
1) quantitative sampling can be realized by operating the first conveyor, the second conveyor and the third conveyor, so that manual sampling is avoided, the error is small, the stability is good, and the accuracy of a fluorescence labeling rate detection result is improved;
2) the sample liquid and the diluting liquid can be obtained by operating the first conveyor and the second conveyor respectively, so that the precision is better than that of a traditional manual sampling mode, and the sample waste is reduced;
3) the filter equipment is used for intercepting the fluorescent marker in the sample liquid, the filter equipment is arranged in the fluorescent marking rate detection system, the interference of the fluorescent marker and impurities can be avoided, the accuracy of the fluorescent marking rate detection result is influenced, and meanwhile, the concentration of the fluorescent monomer in the solution to be detected can be ensured in the detection limit range of the fluorescent detector by the design that the output end of the diluent tank is connected with the second input end of the fixed container through the second conveyor, so that the fluorescent marking rate detection system provided by the application has excellent detection precision.
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without inventive exercise.
FIG. 1 is a schematic diagram of a fluorescence labeling rate detection system provided herein;
reference numerals: 1-a sample tank; 21-a first conveyor; 22-a second conveyor; 23-a third conveyor; 3-a filtration device; 4-determining a container; 5-a fluorescence detector; 6-a diluent tank; 7-a waste liquid tank; 81-a first control valve; 82-a second control valve; 9-stirrer.
Detailed Description
The technical solutions of the embodiments of the present application will be described clearly and completely with reference to the accompanying drawings, and it is obvious that the described embodiments are some, but not all, of the embodiments of the present application. All other embodiments obtained by a person of ordinary skill in the art based on the embodiments in the present application without any creative effort belong to the protection scope of the embodiments in the present application.
In the description of the embodiments of the present application, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, and are only for convenience in describing the embodiments of the present application and simplifying the description, but do not indicate or imply that the referred devices or elements must have specific orientations, be configured in specific orientations, and operate, and thus, should not be construed as limiting the embodiments of the present application. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
In the description of the embodiments of the present application, it should be noted that the terms "mounted," "connected," and "connected" are used broadly and are defined as, for example, a fixed connection, an exchangeable connection, an integrated connection, a mechanical connection, an electrical connection, a direct connection, an indirect connection through an intermediate medium, and a communication between two elements, unless otherwise explicitly stated or limited. Specific meanings of the above terms in the embodiments of the present application can be understood in specific cases by those of ordinary skill in the art.
The embodiment of the application discloses a fluorescence labeling rate detection system.
Referring to fig. 1, the fluorescence mark rate detection system provided in the embodiment of the present application mainly includes a sample tank 1, a filter device 3, a constant volume device 4, a diluent tank 6, and a fluorescence detector 5, an input end of the filter device 3 is connected to an output end of the sample tank 1 through a first conveyor 21, a first input end of the constant volume device 4 is connected to an output end of the filter device 3, a second input end of the constant volume device 4 is connected to an output end of the diluent tank 6 through a second conveyor 22, and an input end of the fluorescence detector 5 is connected to an output end of the constant volume device 4 through a third conveyor 23.
The sample tank 1 is used for providing a sample solution, and a fluorescent marker and a fluorescent monomer for fluorescent labeling are dispersed in the sample solution. It is understood that the fluorescent label is a kind of reaction in which a fluorescent monomer is labeled on an object to be labeled to form a fluorescent label, and the fluorescent monomer is a raw material for forming the fluorescent label, and the fluorescent monomer is modified on the object to be labeled to form the fluorescent label through the fluorescent label reaction.
In one embodiment, the sample tank 1 is a reaction kettle, and the sample liquid is a reaction liquid in the labeling reaction, and the fluorescence labeling rate detection system provided by the present application can realize real-time monitoring of the labeling process, such as quantitative sampling from the reaction kettle at regular time for fluorescence detection, determining the status of the fluorescence labeling reaction according to the monitoring result, and determining when the labeling reaction is terminated.
As the reaction continues, the amount of the fluorescent monomer labeled on the substance to be labeled increases, which causes the concentration of the free fluorescent monomer in the sample solution to decrease, so that the excitation light intensity detected by the fluorescence detector 5 gradually decreases from a highest detection value to a stable detection value, and when the excitation light intensity decreases to the stable detection value, the reaction is completed and the reaction is terminated. And then, converting the stable fluorescence intensity to obtain the fluorescence labeling rate in the fluorescence labeling reaction process, namely the ratio of the number of the fluorescent monomers which are labeled on the to-be-labeled substance to form the fluorescence label to the number of the fluorescent monomers in the system at the initial stage of the reaction.
The fluorescence marker is temperature-sensitive nanogel for marking the fluorescence monomer, namely the to-be-marked object is temperature-sensitive nanogel, while the temperature-sensitive nanogel used in the embodiment of the application is N-isopropyl acrylamide hydrogel, and the temperature-sensitive nanogel can be N-isopropyl acrylamide hydrogel specifically. Specifically, the temperature-sensitive nanogel is N-isopropyl acrylamide or a cross-linked polymer of N-isopropyl acrylamide and a comonomer, the comonomer is preferably at least one of Acrylic Acid (AA), N-N-propyl acrylamide (NNP), methacrylic acid (MAA), hydroxyethyl methacrylate (HEMA), hydroxyethyl acrylate (HEA) and acrylamide (AAm), and the cross-linking agent is N, N-methylene bisacrylamide. The temperature-sensitive nano gel is an embolization material which is discovered by the research and development team at the beginning of the century and has excellent human body temperature response activity and embolization effect.
The first conveyor 21, the second conveyor 22 and the third conveyor 23 are all used as solution conveying devices and can be designed as peristaltic pumps, and quantitative sampling, even ultra-quantitative sampling can be realized by setting flow parameters of the peristaltic pumps.
The first conveyor 21 is used for conveying the sample liquid in the sample tank 1 to the filtering device 3, that is, providing sample liquid conveying power, so as to meet the sample liquid conveying requirement, and meanwhile, the flow data of the first conveyor 21 can be set, so that quantitative conveying is realized. In some embodiments, the first conveyor 21 is a peristaltic pump.
The first conveyor 21 is connected at its input to the output of the sample tank 1 and at its output to the input of the filter device 3. The output end of the sample tank 1 can be arranged at the bottom of the sample tank, so that liquid can be conveniently discharged, the sample tank 1 and the filtering device 3 can be arranged above the filtering device 3 in a relative arrangement position, the sample liquid can be further conveyed by the aid of the gravity of the sample liquid, and the purpose of saving energy consumption is achieved.
The output end of the filtering device 3 is connected with the first input end of the constant volume device 4 and is used for intercepting fluorescent markers in the sample liquid, namely intercepting impurities such as high molecular products in the sample liquid, and fluorescent monomers can enter the constant volume device 4 through the filtering device 3, so that the solution to be detected which is subsequently conveyed to the constant volume device 4 is mainly the fluorescent monomers which are not successfully marked.
The aperture of the filtering hole of the filtering device 3 is adjustable, namely, the filter element can be selected for use as a filter with replaceable design, and the filter with the proper aperture can be selected according to the type of the fluorescent marker. In some embodiments, the filtering device 3 comprises at least two filters, and the filtering pores of each filter have different pore sizes, and preferably, the filters are arranged in parallel between the first conveyor 21 and the constant volume device 4 and share the same input end and output end, so that the filter with the proper filtering pore size can be selected according to the type of the labeled temperature-sensitive nanogel for use. The application range of the system can be enlarged and the applicability can be improved no matter the filter device 3 is designed to be adjustable or the filter device 3 is designed to be selectable.
The diluent tank 6 is used for providing a diluent, the diluent is ultrapure water, PBS buffer solution, physiological saline, animal serum or animal whole blood, and a proper diluent can be flexibly selected according to actual conditions. The above-mentioned diluents are commercially available or can be prepared according to a predetermined formulation by a method conventionally used in the art.
The second conveyor 22 is used for conveying the sample liquid in the diluent liquid tank 6 to the constant volume device 4, similar to the first conveyor 21, the second conveyor 22 also provides solution conveying power, and in practical application, quantitative conveying can be realized by setting flow data of the second conveyor 22. In some embodiments, second conveyor 22 is a peristaltic pump.
The output end of the diluent tank 6 is connected with the input end of the second conveyor 22, the output end of the second conveyor 22 is connected with the second input end of the constant volume device 4, and the second conveyor 22 is used for conveying the diluent in the diluent tank 6 to the constant volume device 4, so that the concentration of the fluorescent monomer subsequently conveyed to the fluorescence detector 5 meets the requirement of the detection limit of the fluorescence detector 5. The relative distribution position between the diluent tank 6 and the constant volume device 4 can be that the diluent tank 6 is arranged above the constant volume device 4, and can also play a role in saving energy consumption for conveying diluent.
The fixed container 4 can be a transparent sealed container, and the surface of the fixed container can be marked with metering scales, so that the operation personnel can conveniently check the amount of the solution to be detected and the diluting liquid entering the fixed container 4. The output of deciding container 4 can locate self bottom, and self lateral wall top can be located to the first input of constant volume ware 4, and self top can be located to the second input of constant volume ware 4, specifically does not do the restriction.
The output end of the fixed container 4 is connected with the input end of a third conveyor 23, the output end of the third conveyor 23 is connected with the input end of the fluorescence detector 5, and the third conveyor 23 is used for conveying the diluted solution to be detected in the fixed container 4. Similarly, the position distribution between the constant volume device 4 and the fluorescence detector 5 can be that the constant volume device 4 is distributed above the fluorescence detector 5, so that the energy consumption of the solution to be detected during conveying can be saved. In some embodiments, third conveyor 23 is a peristaltic pump.
The fluorescence detector 5 is a detector capable of generating fluorescence excitation light with a certain range of wavelengths and receiving fluorescence excitation light with a certain range of wavelengths, and when in use, the emission wavelength and the detection wavelength of the fluorescence detector 5 can be set according to the type of the fluorescence marker. The fluorescence detector is an existing device purchased on the market.
As shown in fig. 1, as for the connection of the sample tank 1, the first conveyor 21, the filtering device 3 and the constant volume device 4, the sample tank, the first conveyor, the filtering device and the constant volume device are connected in sequence through a conveying pipeline; as for the connection of the diluent tank 6, the second conveyor 22 and the constant volume device 4, the connection is specifically realized in sequence through a conveying pipeline; the connection of the constant volume device 4, the third conveyor 23 and the fluorescence detector 5 is specifically connected in sequence by a conveying pipe. The delivery pipe is a pipe for solution circulation, which is a conventional device purchased in the market.
Wherein, still be provided with first control valve 81 on the conveyer pipe that the place between filter equipment 3 and the constant volume ware 4, close this first control valve 81 after the sample liquid quantity that gets into constant volume ware 4 satisfies the detection demand to further improve the quantitative precision of sample liquid. Similarly, a second control valve 82 is further disposed on the delivery pipeline between the second conveyor 22 and the constant volume device 4, and the function of the second control valve 82 is similar to that of the first control valve 81, which is not described in detail. The first control valve 81 and the second control valve 82 may be, for example, a solenoid flow valve, and are not limited.
In some embodiments, the fluorescence labeling rate detection system provided in the embodiments of the present application further comprises: a controller; the controller is electrically connected with the first conveyor 21, the second conveyor 22, the third conveyor 23 and the fluorescence detector 5, and can automatically control the first conveyor 21, the second conveyor 22, the third conveyor 23 and the fluorescence detector 5 by controlling the controller, so that quantitative sampling and detection are realized, even the work is stopped, and thus, automatic real-time sampling and detection are realized. The controller can be a PLC controller or a microprocessor and the like, is not limited, and can realize automatic real-time and quantitative fluorescence labeling rate detection operation by configuring the controller. It will be appreciated that to further increase the degree of automation, the controller is also electrically connected to the first control valve 81 and the second control valve 82.
As shown in fig. 1, the fluorescence labeling rate detection system provided in the embodiment of the present application further includes: and the waste liquid tank 7 is connected with the output end of the fluorescence detector 5, and is used for receiving the detected waste liquid.
According to the technical scheme, the fluorescence labeling rate detection system designed by the application has the advantages that when the fluorescence labeling rate detection system is specifically applied, the sample liquid in the sample tank 1 can be quantitatively conveyed to a fixed container through a first conveyor 21, and the fluorescent marker is intercepted by a filtering device 3 during the conveying, so that the solution to be measured transferred into the fixed container 4 is mainly unlabeled fluorescent monomer, and thereafter, the solution to be measured is diluted by quantitatively feeding the diluting solution in the diluting solution tank 6 to the constant volume device 4 by the second conveyor 22, so that the concentration of the fluorescent monomer subsequently delivered to the fluorescence detector 5 meets the requirement of the detection limit of the fluorescence detector 5, and then, and conveying the diluted solution to be detected to a fluorescence detector 5 through a third conveyor 23 for detection, and converting the detected fluorescence intensity to obtain the fluorescence labeling rate in the fluorescence labeling reaction process.
Compared with the prior art, the fluorescent marker detection system designed by the matching of the quantitative sampling, filtering, diluting and detecting processes has the following beneficial effects:
1) quantitative sampling can be realized by operating the first conveyor 21, the second conveyor 22 and the third conveyor 23, so that manual sampling is avoided, the error is small, the stability is good, and the accuracy of the fluorescence labeling rate detection result is improved;
2) the acquisition of the sample liquid and the dilution liquid can be realized by operating the first conveyor 21 and the second conveyor 22 respectively, so that the precision is better than that of a traditional manual sampling mode, and the sample waste is reduced;
3) filter equipment 3 is arranged in intercepting the fluorescence marker in the sample liquid, sets up filter equipment in fluorescence mark rate detecting system, can avoid fluorescence marker and impurity to disturb, simultaneously, this design that the output of diluent jar 6 is connected through second conveyer 22 and the second input of constant volume ware 4 can guarantee the fluorescence monomer concentration in the solution that awaits measuring in the detection limit scope of fluorescence detector for the fluorescence mark rate detecting system that this application provided has excellent detection precision.
The detection process of the fluorescent marker detection system provided by the embodiment of the application can be as follows:
1. the sample injection volume of each peristaltic pump is set, for example, to 1 mL/min;
2. the proper filtering aperture of the filtering device 3 is determined according to the type of the marked temperature-sensitive nanogel, and the filtering can be realized by replacing a filter element or selecting different filters;
3. setting the emission wavelength and the detection wavelength of the fluorescence detector 5 according to the type of the marked temperature-sensitive nanogel;
4. the first conveyor 21 is started, the first control valve 81 is adjusted to quantitatively convey the sample liquid to the constant volume device 4, for example, to quantitatively convey 10 μ L, and the first conveyor 21 and the first control valve 81 are closed after the conveyance is completed. The second conveyor 22 is started again, the second control valve 82 is adjusted to quantitatively convey the dilution liquid to the constant volume device 4, for example, 990 μ L of the dilution liquid is quantitatively conveyed, the dilution ratio can be 1: 100-200, specifically, the dilution ratio can be set according to the requirement, and the second conveyor 22 and the second control valve 82 are closed after the conveyance is finished. And starting the third conveyor 23, and conveying the proportioned solution to be detected to the fluorescence detector 5 for detection.
While the fluorescence mark rate detecting system provided by the present application has been described in detail, the present disclosure should not be construed as limiting the present application, and those skilled in the art will appreciate that the following detailed description and the following detailed description can be modified according to the spirit and scope of the present application.
Claims (10)
1. A fluorescence labeling rate detecting system, comprising:
the sample tank is used for providing a sample liquid, and a fluorescent marker and a fluorescent monomer for fluorescent labeling are dispersed in the sample liquid;
the diluent tank is used for providing a diluent;
the filter device is used for trapping the fluorescent marker, and the input end of the filter device is connected with the output end of the sample tank through a first conveyor;
a first input end of the fixed container is connected with the output end of the filtering device, and a second input end of the fixed container is connected with the output end of the diluent tank through a second conveyor;
and the input end of the fluorescence detector is connected with the output end of the fixed container through a third conveyor.
2. The fluorescence labeling rate detecting system of claim 1, wherein the sample tank is a reaction kettle; and/or
The fluorescent marker is temperature-sensitive nanogel marked with the fluorescent monomer, and the temperature-sensitive nanogel is N-isopropyl acrylamide hydrogel.
3. The fluorescence mark rate detection system of claim 1, wherein the first conveyor is a peristaltic pump; and/or
The second conveyor is a peristaltic pump; and/or
The third conveyor is a peristaltic pump.
4. The fluorescence labeling rate detecting system of claim 1, further comprising: a controller;
the controller is electrically connected to the first conveyor, the second conveyor, the third conveyor, and the fluorescence detector.
5. The fluorescence labeling rate detecting system according to any one of claims 1 to 4, wherein the sample tank, the first conveyor, the filtering means, and the fixed container are connected in sequence by a conveying pipe; and/or
The diluent tank, the second conveyor and the fixed container are sequentially connected through a conveying pipeline; and/or
The fixed container, the third conveyor and the fluorescence detector are sequentially connected through a conveying pipeline.
6. The fluorescence mark rate detecting system according to claim 5, wherein a first control valve is further disposed on the delivery line between the filtering device and the constant volume device; and/or
And a second control valve is also arranged on the conveying pipeline between the second conveyor and the constant volume device.
7. The fluorescence labeling rate detecting system according to any one of claims 1 to 4, wherein the aperture of the filter pores of the filter device is adjustable; and/or
The filtering device comprises at least two filters, and the filter pores of the filters are different in pore size.
8. The fluorescence labeling rate detecting system according to any one of claims 1 to 4, wherein the diluting solution is ultrapure water, PBS buffer, physiological saline, animal serum or animal whole blood.
9. The fluorescence labeling rate detecting system according to any one of claims 1 to 4, wherein a stirrer is further connected to the sample tank, and the stirrer is provided with a stirring portion for stirring the sample liquid.
10. The fluorescence labeling rate detecting system according to any one of claims 1 to 4, further comprising: a waste liquid tank;
the waste liquid tank is connected with the output end of the fluorescence detector.
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