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

Confining liquid and application thereof

A technology of blocking liquid and solution, which is applied in the direction of instruments, analytical materials, particles and sedimentation analysis, etc., can solve the problems of perishable, poor sealing effect of blocking liquid, etc., and achieve the effect of not easy to deteriorate, saving experimental cost, and accurate results

Inactive Publication Date: 2020-01-03
GUANGDONG UNIV OF TECH
View PDF11 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The invention provides a sealing liquid and its application, which solves the problems of poor sealing effect and easy deterioration of the existing sealing liquid

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
  • Confining liquid and application thereof
  • Confining liquid and application thereof
  • Confining liquid and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] This example is the induction of human induced pluripotent stem cells (hiPSCs) into cardiomyocytes

[0044]1. HiPSC-U1 culture method: freeze-thaw Matrigel from -80°C to 4°C overnight, aliquot Matrigel the next day and add 100 μl Matrigel to 10ml DMEM / F-12 for culture in a 15ml centrifuge tube, store Store in the refrigerator at 4°C for later use. Every day, replace each well with new medium: 4ml TeSR-E8 medium until the number of hiPSCs reaches about 75%.

[0045] 2. On the 0th day of differentiation, discard the old medium, replace and add 4ml (12μM CHIR99021+RPMI / B-27minus insulin) medium to each well, and return the multi-well plate to 37°C, 5% CO 2 Incubate in the incubator for 24h.

[0046] 3. On the first day of differentiation, discard the old medium, add 4ml of RPMI / B-27 medium (without insulin) to each well, and return the multi-well plate to 37°C, 5% CO 2 Incubate in the incubator for 24h.

[0047] 4. On the third day of differentiation, 72 hours after ad...

Embodiment 2

[0051] This embodiment is the immunofluorescent staining of cardiomyocytes in Example 1

[0052] (1) Digestion:

[0053] Aspirate off the old medium and wash the differentiated cells twice with 2 ml of PBS per well. Aspirate the PBS, add 2ml (0.25% (wt / vol) trypsin-EDTA) dissociating enzyme to dissociate the cells, and digest at 37°C for 5min in a 5% incubator.

[0054] (2) filter cells:

[0055] Cells were singulated by pipetting 5-10 times with a 1 ml pipette. The pipetted single cell suspension was then transferred to a 15ml centrifuge tube containing 4ml RPMI 20, and the cells were naturally filtered by gravity with a 70μm cell sieve.

[0056] (4) Counting:

[0057] Take 10 μl of the cell suspension and count the cells with a hemocytometer; and centrifuge the cell suspension at a speed of 1000 r / min for 4 min at room temperature. After centrifugation, the supernatant was discarded using a 1 ml pipette.

[0058] (5) Cell fixation:

[0059] After the old medium was di...

Embodiment 3

[0070] In this example, flow cytometric analysis was performed on the cardiomyocytes after immunofluorescence staining in Example 2 and Comparative Example 1.

[0071] The cardiomyocytes incubated in Example 2 and Comparative Example 1 were washed twice with 2 ml FlowBuffer-2 respectively; the cell pellet was resuspended in 50 μl FlowBuffer-1, and transferred to a flowing round bottom tube. Place the flow tubes on ice and perform flow cytometric analysis with a FACS Calibur. The result is as Figure 1~6 .

[0072] Depend on Figure 1~3 and Figure 4~6 Comparative analysis shows that: in comparative example 1, the number of cells obtained by blocking liquid injection is relatively small, and the fluorescence detection value is low, so the amount of cells required for the experiment will deviate from the theoretical value of the experiment; in example 2, the number of cells obtained by blocking liquid injection is more than expected. There is little difference between the ex...

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

The invention relates to the technical field of confining liquid, in particular to confining liquid and application thereof. The invention discloses confining liquid. The confining liquid comprises first confining liquid and second confining liquid; the first confining liquid comprises Twain-20, donkey serum, bovine serum albumin and a phosphate buffer solution; the second confining liquid comprises Twain-20, donkey serum, bovine serum albumin, a phosphate buffer solution and polyethylene glycol octyl phenyl ether. According to the confining liquid provided by the invention, the condition thatFcR on a cell surface can be non-specifically combined with the Fc segment of a fluorescein coupling antibody effectively be reduced, and the confining effect is good, so that the result of flow cytometry is more accurate, and the confining liquid is unlikely to go bad.

Description

technical field [0001] The invention relates to the technical field of blocking liquid, in particular to a blocking liquid and its application. Background technique [0002] Flow cytometry is an analytical technique that can perform multiparameter, rapid quantitative analysis of single cells or other biological particles. It can analyze tens of thousands of cells at high speed, and can measure multiple parameters from one cell at the same time. It has the advantages of high speed, high precision and good accuracy. It is one of the most advanced cell quantitative analysis technologies in the contemporary era. Quantitative analysis of cardiomyocytes by flow cytometry can obtain data information on the differentiation of human induced pluripotent stem cells (hiPSCs) into cardiomyocytes, so as to understand the differentiation of cardiomyocytes and provide data support for the follow-up work of culturing cardiomyocytes . [0003] Most of the fluorescein-conjugated antibodies t...

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
Patent Type & Authority Applications(China)
IPC IPC(8): G01N15/14G01N21/64G01N33/533G01N33/537
CPCG01N15/1436G01N21/6428G01N33/533G01N33/537G01N2015/144G01N2021/6439
Inventor 苗小敏汤亚东张焜蓝兴梓陈巧桐陈家盈梁大锡罗竣仁
Owner GUANGDONG UNIV OF TECH
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