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Method for detecting platelet microparticles

A technology for platelet particles and platelets, which is used in measuring devices, individual particle analysis, particle and sedimentation analysis, etc. It can solve the problems of difficult quantification, difficulty, and many interference factors, and achieve the effect of improving accuracy and saving consumption.

Inactive Publication Date: 2014-03-19
GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, electron microscopy needs to cure and spray gold on PMP to achieve the conductivity of PMP, so the preparation of detection samples is cumbersome, difficult, and difficult to quantify
When flow cytometry uses PMP size and surface antigen characteristics to study the formation of PMPs, particles with a diameter of less than 0.5 μm are generally regarded as PMPs, but particles within this range are likely to be other blood cell debris or impurity particles, so it is necessary to Use a variety of platelet-specific fluorescent antibodies to further identify PMPs. However, too many fluorescent antibodies are not only expensive, but also have many interference factors, resulting in deviations in PMP detection results.

Method used

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  • Method for detecting platelet microparticles
  • Method for detecting platelet microparticles

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Experimental program
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Embodiment 1

[0019] A method for detecting platelet particles, comprising the steps of:

[0020] (1) Preparation of platelet samples

[0021] Take 2 ml of venous blood from a healthy person and put it into a sodium citrate anticoagulant tube, centrifuge at 150×g for 15 minutes to obtain platelet-rich plasma; take 200 μL of the platelet-rich plasma and place it in a tube coated with 200 μL of human fibrinogen at a concentration of 2 mg / mL Incubate at room temperature in 35mm cell culture dish for 30 minutes, use PBS buffer to elute the platelets that are not adhered to the bottom of the cell culture dish, add 1.5mL PBS buffer and set aside;

[0022] (2) Take 200 μL of the supernatant in the cell culture dish, and use flow cytometry to quantitatively analyze the number of platelet particles (see figure 2 A);

[0023] (3) Place the Ag / AgCl reference electrode connected to the scanning ion conductance microscope on the cell culture dish obtained in step (1); place the detection electrode co...

Embodiment 2

[0028] A method for detecting platelet particles, comprising the steps of:

[0029] (1) Preparation of platelet samples

[0030] Take 2 ml of venous blood from a healthy person and put it into a sodium citrate anticoagulant tube, centrifuge at 150×g for 15 minutes to obtain platelet-rich plasma; take 200 μL of the platelet-rich plasma and place it in a tube coated with 200 μL of human fibrinogen at a concentration of 2 mg / mL Incubate at room temperature for 20 minutes in a 35mm cell culture dish, use PBS buffer to elute the platelets not adhered to the bottom of the cell culture dish, add 2mL of PBS buffer and set aside;

[0031] (2) Take the supernatant in 200 μL cell culture dish, and use flow cytometry to quantitatively analyze the number of platelet particles;

[0032] (3) Place the Ag / AgCl reference electrode connected to the scanning ion conductance microscope on the cell culture dish obtained in step (1); place the detection electrode connected to the scanning ion cond...

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Abstract

The invention discloses a method for detecting platelet microparticles. The method comprises the following steps of: (1) preparing a platelet sample; (2) quantitatively analyzing the number of the platelet microparticles; (3) placing a reference electrode and a detection electrode which are connected to a scanning ionic conductance microscope in a culture dish; (4) drawing a three-dimensional topological structure chart of the platelet surface topography; (5) drawing a three-dimensional topological structure chart of the platelet surface topography 60 minutes after adding an activator, and qualitatively observing the formation of the platelet microparticles; and (6) taking the supernate in the cell culture dish with the activator, and quantitatively analyzing the number of the platelet microparticles formed after the platelet is activated. The method disclosed by the invention avoids the influence of other blood cell debris and contamination particles on the detection of platelet microparticles (PMP), qualitatively and quantitatively detects the PMP, does not need dyeing or any special treatment, saves the usage of platelet specific antibodies, and improves the accuracy of PMP formation detection.

Description

technical field [0001] The invention relates to a method for detecting platelet microparticles. Background technique [0002] Blood cell microparticles (Microparticles, MP) are a kind of ultrafine particles that exist in the blood and are released by stimulation, activation or apoptosis of various cells (such as platelets, white blood cells, lymphocytes, red blood cells, endothelial cells, vascular smooth muscle cells, etc.). membranous vesicles. Among them, platelet microparticles (Platelet Microparticles, PMPs) account for about 70% to 90% of the total MP in blood. The platelet particle (PMP) membrane carries most of the active ingredients on the platelet membrane in the resting state, so it has the function of promoting hemostasis and accelerating coagulation like platelets; in addition, the PMP membrane also carries the markers on the activated platelet membrane, which not only has Strong procoagulant activity, also has certain anticoagulant activity, plays an importan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N15/14
Inventor 张彦军张建宁董京飞刘丽
Owner GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV