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Preparation method and application of exosome for nanoflow cytometry detection

A flow cytometer and exosome technology, applied in the field of exosome preparation, can solve the problems of difficult promotion and high cost, and achieve the effects of high sensitivity, high selectivity and simple operation.

Pending Publication Date: 2021-06-04
THE FIRST AFFILIATED HOSPITAL OF BENGBU MEDICAL COLLEGE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Now based on the high cost of some immunosorbent methods, it is difficult to promote, while the indirect labeling flow cytometry technology is based on the improvement of ordinary flow technology, and its application opportunities are more

Method used

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  • Preparation method and application of exosome for nanoflow cytometry detection
  • Preparation method and application of exosome for nanoflow cytometry detection
  • Preparation method and application of exosome for nanoflow cytometry detection

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Embodiment 1

[0023] A preparation method for exosomes detected by nanometer flow cytometry, comprising the steps of:

[0024] (1) Take exosome samples;

[0025] (2) The exosome samples were thawed on ice;

[0026] (3) Ultrasonic treatment of exosome samples;

[0027] (4) Add PBS to the centrifuge tube, then add the antibody to the centrifuge tube, mix thoroughly, then add the exosome sample, and then obtain the exosome resuspended in PBS after incubation and centrifugation in an ultra-high speed centrifuge.

[0028] During the sonication in step (3), the duration of sonication is controlled to be 10 min.

[0029] The capacity of the centrifuge tube in step (4) is 200ul; the amount of PBS added is 48ul; the amount of antibody added is 1ul; the amount of exosome sample added is 1ul.

[0030] The length of incubation in step (4) is controlled to be 30min.

[0031] The duration of centrifugation in the ultrahigh speed centrifuge in step (4) is 20min, and the centrifugal force is controlled...

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Abstract

The invention discloses a preparation method of an exosome for nanoflow cytometry detection, and belongs to the technical field of cell substance detection. The preparation method comprises the following steps: (1) taking an exosome sample; (2) carrying out on-ice unfreezing treatment on the exosome sample; (3) performing ultrasonic treatment on the exosome sample; and (4) adding PBS into a centrifugal tube, then adding an antibody into the centrifugal tube, fully and uniformly mixing the mixture, then adding the exosome sample, performing incubating, and carrying out centrifugal treatment by an ultra-high-speed centrifuge to obtain the PBS resuspended exosome. The invention discloses the preparation method of the exosome for nanoflow cytometry detection. The method is simple to operate, accurate, reliable, high in sensitivity, free of extraction by an organic solvent and high in selectivity to a target detection object.

Description

technical field [0001] The invention belongs to the technical field of cell substance detection, and in particular relates to a preparation method and application of exosomes for nano flow cytometer detection. Background technique [0002] The working principle of flow cytometry is to conduct multi-parameter and rapid quantitative analysis of single cells or other biological particles through monoclonal antibodies at the molecular level. Flow cytometry generally includes three main steps: fluorescent antibody incubation, centrifugation and resuspension, and on-machine detection. Exosomes refer to small membrane vesicles (30-150nm) that contain complex RNA and proteins and are secreted autonomously by cells. Now they specifically refer to disc-shaped vesicles with a diameter of 40-100nm, which carry the above information and play a role in cell communication. In recent years, exosomes have shown an important position in the field of tumor research, and the detection of exoso...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N15/14
CPCG01N33/56966G01N15/14G01N15/1484G01N2469/10
Inventor 江畅金海振朱雪茹夏武艳余佳桦高翔傅小龙
Owner THE FIRST AFFILIATED HOSPITAL OF BENGBU MEDICAL COLLEGE
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