CN212568483U

CN212568483U - In-vivo heparin real-time monitoring device based on micro-fluidic chip

Info

Publication number: CN212568483U
Application number: CN202021836579.2U
Authority: CN
Inventors: 丁涛; 周星贝; 王树水
Original assignee: South China University of Technology SCUT
Current assignee: South China University of Technology SCUT
Priority date: 2020-08-28
Filing date: 2020-08-28
Publication date: 2021-02-19
Anticipated expiration: 2030-08-28

Abstract

The utility model discloses an in vivo heparin real-time monitoring device based on a microfluidic chip, which comprises a microdialysis probe used for acquiring a sample to be tested; the micro-fluidic chip is used for fully mixing a sample to be detected and a detection reagent therein; the fluorescence measuring instrument is arranged at the tail end of the microfluidic chip and is used for observing a fluorescence measuring value obtained after the sample and the detection reagent are mixed; and the data processing unit is connected with the fluorescence measuring instrument and obtains the heparin concentration through a fluorescence measuring value. The device has the advantages of high sensitivity and high analysis speed.

Description

In-vivo heparin real-time monitoring device based on micro-fluidic chip

Technical Field

The utility model relates to a detection area, concretely relates to internal heparin real-time supervision device based on micro-fluidic chip.

Background

Heparin is a commonly used clinical anticoagulant. During the treatment process, the blood concentration of heparin is a very important therapeutic effect indicating tool, and excessive concentration can cause bleeding, low concentration and insufficient anticoagulation often cause thrombus, thereby causing serious consequences. Therefore, there is a need for close monitoring of drug concentrations in patients when heparin is used. Currently, the clinical heparin monitoring items are Activated Partial Thromboplastin Time (APTT) and Activated Clotting Time (ACT). These tests are generally performed at 37 ℃ and the coagulation status of patients after heparin application is determined by adding negatively charged foreign substances (e.g., glass, kaolin, sulfate, collagen, etc.) to the plasma/whole blood sample to activate the coagulation process and measuring the time required for the sample to coagulate. APTT and ACT do not directly measure heparin concentration in blood and due to limitations in the measurement principle, these methods are not applicable to blood samples using large doses of heparin. The measurement results of the APTT and the ACT are easily influenced by various external factors such as blood drawing amount, inspection time, storage temperature and the like, and the sensitivity and the accuracy are poor; the inspection and measurement process consumes long time, the result is reported slowly, and the timeliness is insufficient; blood needs to be collected again in each detection, and the harm to patients is large. Therefore, it is necessary to develop a device for monitoring heparin blood concentration with small trauma.

SUMMERY OF THE UTILITY MODEL

In order to overcome the shortcoming and the deficiency that prior art exists, the utility model provides an internal heparin real-time supervision device based on micro-fluidic chip, this device with microdialysis and micro-fluidic technology combined application, have automatic and developments survey in succession, advantage that the wound is little.

The utility model adopts the following technical scheme:

an in vivo heparin real-time monitoring device based on a microfluidic chip comprises,

the microdialysis probe is used for obtaining a sample to be detected;

the micro-fluidic chip is used for fully mixing a sample to be detected and a detection reagent therein;

the fluorescence measuring instrument is arranged at the tail end of the microfluidic chip and is used for observing a fluorescence measuring value obtained after the sample and the detection reagent are mixed;

and the data processing unit is connected with the fluorescence measuring instrument and obtains the heparin concentration through a fluorescence measuring value.

The detection reagent comprises FXa and a fluorescent reagent.

Further, the device also comprises a microdialysis perfusion pump, wherein the microdialysis perfusion pump slowly injects phosphate buffer salt solution into the microdialysis probe at 400 nl/min.

The micro-fluidic chip comprises a liquid inlet, a flow channel, an observation position and a waste liquid port, wherein one end of the flow channel is connected with the liquid inlet, the other end of the flow channel is connected with the waste liquid port, and the observation position is arranged at the tail end of the flow channel.

Further, the device also comprises an injection pump, and the injection pump is communicated with a liquid inlet of the microfluidic chip.

The utility model has the advantages that:

the device combines the microdialysis and the microfluidic technology, and simultaneously, the heparin concentration in blood can be measured in real time by combining an anti-FXa method and fluorescence quantitative detection, so that on-line monitoring is realized;

the device also has the advantages of high sensitivity and high analysis speed.

Drawings

FIG. 1 is a schematic diagram of the apparatus of the present invention;

fig. 2 is a schematic diagram of the structure of the microfluidic chip of the present invention.

Detailed Description

The present invention will be described in further detail with reference to the following examples and drawings, but the present invention is not limited thereto.

Examples

As shown in fig. 1, an in vivo heparin real-time monitoring device based on a microfluidic chip comprises:

and the microdialysis probe 2 is used for obtaining a sample to be tested, the microdialysis probe is immersed in a heparin solution or blood 3 of an individual to be tested, the microdialysis perfusion pump 1 is started, and phosphate buffer saline solution is slowly injected into the microdialysis probe at the speed of 400nl/min for sampling.

The microfluidic chip 5 is used for fully mixing a sample to be detected and a detection reagent; the detection reagent is FXa and a fluorescent reagent.

The method specifically comprises the following steps: the microdialysis blood sample is injected into the microfluidic chip, the FXa and the fluorescent reagent are automatically injected into the chip by using the injection pump 4, the fluorescent reagent can release fluorescence after being decomposed by the FXa, and the heparin in the sample can inhibit the decomposition reaction of the FXa on the fluorescent reagent. Therefore, the strength of fluorescence released by the sample after being mixed with the two reagents can quantitatively reflect the content of heparin in the sample, and the fluorescence signal value is inversely proportional to the concentration of the heparin. After entering the chip, the blood sample is fully mixed with the reagent in the chip flow channel and stably releases fluorescence, when the mixture flows through the observation position of the chip, the fluorescence released by the mixture is measured by using the fluorescence measuring instrument 6, and the concentration of heparin is calculated according to the fluorescence signal value. Thereby realizing the on-line monitoring of heparin in the body of the subject and evaluating the anticoagulation condition in real time.

As shown in fig. 2, the microfluidic chip comprises a liquid inlet 5-1, a flow channel 5-2, an observation position 5-3 and a waste liquid port 5-4, wherein one end of the flow channel is connected with the liquid inlet, the other end of the flow channel is connected with the waste liquid port, and the observation position is provided with the tail end of the flow channel.

The fluorescence measuring instrument is used for observing a fluorescence measuring value of a mixed sample and a detection reagent through an observation position, and recording sequential fluorescence signal values at intervals, wherein the fluorescence detection excitation wavelength is 350nm, and the emission wavelength is 450 nm;

the data processing unit is connected with the fluorescence measuring instrument and obtains the heparin concentration through a fluorescence measuring value, and the data processing unit specifically comprises: and calculating the concentration of heparin in the hemodialysis sample of the experimental specimen according to a standard curve method, and drawing a blood concentration-time change curve of the specimen after heparin injection according to the result to determine the concentration change condition of the heparin in the specimen.

In this example, 11Elite, Harvard Apparatus, USA, and CMA 20, Sweden CMA was used as the syringe pump. The microfluidic chip is made of Guangzhou chrome company, the fluorescence measuring instrument is made of Axio Imager 2 and Germany Zeiss company, the phosphate buffer solution is made of Sigma-Aldrich company in the United states, the FXa is made of France Hyphen Biomed company, and the FXa specific fluorescent substrate is made of Pentapharm company in Switzerland.

The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited by 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

1. An in vivo heparin real-time monitoring device based on a microfluidic chip is characterized by comprising,

the microdialysis probe is used for obtaining a sample to be detected;

2. The in vivo heparin real-time monitoring device according to claim 1, wherein the detection reagent comprises FXa and a fluorescent reagent.

3. The in vivo heparin real-time monitoring device according to any one of claims 1-2, wherein further comprising a microdialysis perfusion pump which slowly injects phosphate buffered saline solution into the microdialysis probe at 400 nl/min.

4. The in vivo heparin real-time monitoring device according to claim 1, wherein the microfluidic chip comprises a liquid inlet, a flow channel, an observation position and a waste liquid port, one end of the flow channel is connected with the liquid inlet, the other end of the flow channel is connected with the waste liquid port, and the observation position is arranged at the tail end of the flow channel.

5. The in vivo heparin real-time monitoring device according to claim 4, further comprising an injection pump, wherein the injection pump is communicated with the liquid inlet of the microfluidic chip.