Method for separating single cells in animals and plants

A separation method and single-cell technology, applied in the field of single-cell separation in animals and plants, can solve the problems of low cell purity, impurity separation, small diameter, etc., to improve cell yield and purity, promote cell activity, and high cell purity. Effect

Pending Publication Date: 2020-09-11
INNER MONGOLIA AUTONOMOUS REGION ACAD OF AGRI & ANIMAL HUSBANDRY SCI
View PDF5 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, multiple cells and proteins in animal and plant tissues stick together, and the viability of single cells is far inferior to that of microorganisms, so it is also difficult to separate impurities
[0004] To sum up, the problem existi

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
  • Method for separating single cells in animals and plants
  • Method for separating single cells in animals and plants
  • Method for separating single cells in animals and plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A method for isolating single cells in animals and plants, comprising:

[0026] Step 1, biological sample pretreatment: add active liquid and separation enzyme to the biological sample, soak for 1 hour, and obtain the soaking mixture;

[0027] The active liquid was prepared according to the following method: sodium chloride 9g, potassium chloride 0.5mg, lanthanum nitrate 0.01mg, 4-hydroxyethylpiperazineethanesulfonic acid 2.4g, distilled water to 1L, pH 7.2;

[0028] The solid-liquid ratio of the biological sample and the active liquid is 10g:100mL;

[0029] The isolated enzyme is a mixture of pectinase and collagenase, and 0.5 g of pectinase and 0.5 g of collagenase are added to every liter of active liquid;

[0030] Step 2, small molecule filtration: filter the soaking mixture with a filter membrane with a pore size of 0.22 μm;

[0031] Step 3, large particle filtration: soak and stir the filter membrane and the sediment above it in the active solution, resuspend si...

Embodiment 2

[0034] A method for isolating single cells in animals and plants, comprising:

[0035] Step 1, biological sample pretreatment: add active liquid and separation enzyme to the biological sample, soak for 1.5h, and obtain the soaking mixture;

[0036] The active liquid was prepared according to the following method: 8.5g of sodium chloride, 0.7mg of potassium chloride, 0.05mg of lanthanum nitrate, 2.4g of 4-hydroxyethylpiperazineethanesulfonic acid, distilled water to 1L, pH 7.4;

[0037] The solid-liquid ratio of the biological sample and the active liquid is 15g:100mL;

[0038] The isolated enzyme is a mixture of pectinase and collagenase, and 0.75 g of pectinase and 0.75 g of collagenase are added per liter of active liquid;

[0039] Step 2, small molecule filtration: filter the soaking mixture with a filter membrane with a pore size of 0.45 μm;

[0040] Step 3, large particle filtration: soak and stir the filter membrane and the sediment above it in the active solution, res...

Embodiment 3

[0043] A method for isolating single cells in animals and plants, comprising:

[0044] Step 1, biological sample pretreatment: add active liquid and separation enzyme to the biological sample, soak for 2 hours, and obtain the soaking mixture;

[0045] The active solution was prepared according to the following method: sodium chloride 9g, potassium chloride 0.5mg, lanthanum chloride 0.03mg, 4-hydroxyethylpiperazineethanesulfonic acid 2.4g, distilled water to 1L, pH 7.2;

[0046] The solid-liquid ratio of the biological sample and the active liquid is 20g:100mL;

[0047] The isolated enzyme is a mixture of pectinase and collagenase, and 1 g of pectinase and 1 g of collagenase are added in every liter of active liquid;

[0048] Step 2, small molecule filtration: filter the soaking mixture with a filter membrane with a pore size of 0.22 μm;

[0049] Step 3, large particle filtration: soak and stir the filter membrane and the sediment above it in the active solution, resuspend si...

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

PropertyMeasurementUnit
Particle sizeaaaaaaaaaa
Login to view more

Abstract

The invention belongs to the technical field of biology, and particularly relates to a method for separating single cells in animals and plants. The method comprises the steps of adding an active solution and a separating enzyme into a biological sample, and soaking to obtain a soaked mixture, wherein the active solution comprises sodium chloride, potassium chloride, a lanthanum compound and 4-hydroxyethylpiperazine ethanesulfonic acid; filtering the soaked mixture with a filter membrane; soaking and stirring the filter membrane and the precipitate on the filter membrane in the active solutionto enable single cells and microorganisms to be resuspended in the active solution, and collecting a composite cell suspension; and putting glass beads into the composite cell suspension, oscillating, and collecting the single-cell suspension. The cell activity is maintained by adopting the active solution, and the gelatinous substance in the sample is separated by adopting the separating enzyme,so that the cell yield and the cell purity are improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for separating single cells in animals and plants. Background technique [0002] Biological resources are important resources for studying the origin, variation and development direction of organisms. Whether it is a single-celled organism or a single cell isolated from a biological tissue, it has important research value. Typical single-cell biological resources include bacteria, fungi, archaea, etc., and the number and types of these microbial cells are various. Functional microorganisms of different species are widely used in food fermentation, organic matter degradation and other fields. Many functional microbial single cells exist in animal and plant tissues, mixed with other impurities such as protein and fat, and it is difficult to separate impurities. [0003] Living substances such as animals and plants are advanced life forms composed of many single c...

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): C12N5/04C12N5/071C12N1/02
CPCC12N5/04C12N5/067C12N1/02C12N2509/00
Inventor 刘永斌何江峰王力伟祁云霞马跃军付绍印何小龙王标达赖
Owner INNER MONGOLIA AUTONOMOUS REGION ACAD OF AGRI & ANIMAL HUSBANDRY SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap