Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Isotonic hemolysin and preparation method thereof, method for treating biological sample and method for detecting leukocyte membrane antigen

A biological sample, hemolysin technology, applied in the field of immunological cell analysis, can solve the problems of inability to achieve hemolysin effect, differences in white blood cell morphology, differences in granulocyte grouping, etc., to reduce scientific research and medical costs, less cell debris, and easier saved effect

Active Publication Date: 2019-12-17
GENERAL HOSPITAL OF NUCLEAR IND
View PDF3 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The formula of hemolysin has a certain relationship with the instrument. Different instruments use different hemolysin. The same hemolysin shows different effects on different instruments. The main reason is that the leukocyte morphology after hemolysin lysis is different. The optical path design will lead to differences in lymphoid, monocyte and granulocyte grouping
At present, many hemolysins on the market are only suitable for a certain type of flow cytometer, and cannot achieve the role of a hemolysin universal multi-type flow cytometer

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Isotonic hemolysin and preparation method thereof, method for treating biological sample and method for detecting leukocyte membrane antigen
  • Isotonic hemolysin and preparation method thereof, method for treating biological sample and method for detecting leukocyte membrane antigen
  • Isotonic hemolysin and preparation method thereof, method for treating biological sample and method for detecting leukocyte membrane antigen

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0030] On the basis of the above-mentioned isotonic hemolysin, the present invention also discloses a preparation method of isotonic hemolysin, comprising the following steps:

[0031] S1. Take sodium nitrite, sodium chloride, calcium chloride and magnesium chloride according to the above content and add them into the water medium to fully dissolve to obtain a mixed solution.

[0032]S2. Add formaldehyde, glycerol, and isobutanol solution to the mixed buffer solution obtained in step S1, add aqueous medium to 1 liter, adjust the pH value to 7.4-7.6 with sodium bicarbonate, and then filter to obtain hemolysin.

[0033] The method for treating biological samples with the above isotonic hemolysin is as follows: adding the hemolysin to the biological sample at room temperature, and mixing for 10-15 minutes; the biological sample is an anticoagulant venous whole blood; The volume ratio of the hemolysin to the biological sample is 1:40-3:40.

[0034] Meanwhile, the method for detec...

Embodiment

[0049]

[0050] Accuracy test:

[0051] In order to measure the inaccuracy in the results and evaluate the size of the systematic error, the following experiments were carried out: 15 anticoagulated peripheral blood samples from healthy and sub-healthy people were selected, each sample was measured twice with two hemolysins, and the experimental results were recorded. , perform linear regression and correlation analysis. Table 1 Correlation analysis of the percentage of T lymphocyte subsets detected by self-made hemolysin and OptiLyse C hemolysin (%)

[0052]

[0053]

[0054] Precision test:

[0055] In order to evaluate the size of the random error in the measurement results, the following experiments were carried out: 1. Repeated experiments within a batch: using Immuno-Trol cells quality control blood, two kinds of hemolysins were used to add samples for 10 consecutive times in the same time period; 2. batch testing Repeated experiments: Using Immuno-Trol cells ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

An isotonic hemolysin comprises an oxidation dissolving agent, a cosolvent, a cell solubilizer, a cell fixing agent, inorganic salts and water. The oxidation dissolving agent is sodium nitrite, and the content of the oxidation dissolving agent is 2-20g / L. The cosolvent comprises one or more of glycerol, diethylene glycol and propylene glycol, and the content of the cosolvent is 0.3-3mmol / L. The cell solubilizer comprises one or more of isobutanol, methanol and ethanol, and the content of the cell solubilizer is 0.1-1mol / L. The cell fixing agent comprises one or more of paraformaldehyde, formaldehyde, ethanol and acetone, and the content of the cell fixing agent is 10-40g / L. The inorganic salts comprise sodium chloride, magnesium chloride and calcium chloride, wherein the content of the sodium chloride is 1-10g / L, the content of the magnesium chloride is 1-100mmol / L, and the content of the calcium chloride is 1-100mmol / L. A preparation method of the hemolysin, a method for treating a biological sample by the hemolysin and a method for detecting a leukocyte membrane antigen by a flow cytometry are presented. After a sample is treated by the hemolysin, each population of leukocytes ispartitioned obviously, leukocyte membrane protein is not damaged, the original biological activity is retained, and red blood cells can be completely split. The result is accurate, and the performance is stable.

Description

technical field [0001] The invention relates to the field of immunological cell analysis, in particular to an isotonic hemolysin and a preparation method thereof, a method for processing biological samples and a method for detecting white blood cell membrane antigens. Background technique [0002] Blood cell analysis is of great value for scientific research, disease prevention, diagnosis and treatment, from the initial study of cell shape, size, quantity to the most recent antigenic composition on the cell surface. Traditional cellular immunity and non-specific immune function detection techniques cannot perform multi-parameter and high-sensitivity analysis on the size, shape, plasma membrane, and internal structure of target cells from the single-cell level. Flow cytometry can quickly measure, sort, Store and display a series of important biophysical and biochemical characteristic parameters of dispersed cells suspended in liquid, and can sort out specified cell subpopulat...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N33/533G01N15/14
CPCG01N33/533G01N15/1404G01N15/1434
Inventor 俞秋兴
Owner GENERAL HOSPITAL OF NUCLEAR IND
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com