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

Pretreatment solution, kit and pretreatment method for fluorescence in situ hybridization of cell samples

A technique of fluorescence in situ hybridization and pretreatment liquid, which is applied in the fields of biochemical equipment and methods, and the determination/inspection of microorganisms, which can solve the problems of decreased activity and incomplete enzymatic digestion.

Active Publication Date: 2022-05-20
JIAXING ACCB DIAGNOSTICS
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Different protease batches have different activities, and after long-term use, the activity will gradually decrease, which can lead to incomplete enzyme digestion

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
  • Pretreatment solution, kit and pretreatment method for fluorescence in situ hybridization of cell samples
  • Pretreatment solution, kit and pretreatment method for fluorescence in situ hybridization of cell samples
  • Pretreatment solution, kit and pretreatment method for fluorescence in situ hybridization of cell samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] After cytological preparation with normal human blood, pretreatment was carried out according to the method of the present invention (the steps shown in Table 1). postnatal results. After pretreatment, observe under a bright-field microscope whether there are still cell outlines. Then FISH hybridization and DAPI staining were performed. Observe under a fluorescent microscope whether there are DAPI-stained nuclei, and the signal points of the orange gene-specific probe and the green chromosome CEP probe.

[0051] The test results are shown in Table 1 and figure 1 with figure 2 As shown, among them, figure 1 It shows the profile of normal human blood cell droplet under bright field microscope after pretreatment with 100mM NaOH, 0.5% Triton x-100 solution at different temperatures; figure 2 Shown is the FISH signal of the normal human blood cell droplet under the fluorescence microscope after pretreatment with 100mM NaOH, 0.5% Triton x-100 solution.

[0052] figur...

Embodiment 2

[0057] In Example 2, the same blood cell sample as in Example 1 was used for fluorescence in situ hybridization, except that the steps shown in Table 2 were used for pretreatment, and more experimental conditions were tried. The test results are shown in Table 2.

[0058] Table 2:

[0059]

[0060]

[0061]

[0062]It can be seen from Table 2 that after the blood cells are treated with the pretreatment solution containing 15-150 mM lye and 0.1%-1% non-ionic surfactant, FISH signals can be detected. Wherein, the fluorescence in situ detection effect of the blood cells after the pretreatment solution containing 100mM KOH and 0.5% NP-40 is equivalent to the treatment effect of the pretreatment solution containing 100mM NaOH and 0.5% TritonX-100 in Example 1, in The fluorescence images at 25°C are relatively clear. The second is the pretreatment solution containing 15mM NaOH and 0.8% Triton X-100, and the pretreatment solution containing 150mM NaOH and 0.3% Triton X-100...

Embodiment 3

[0064] According to the conventional method, exfoliated cells in the urine of normal people were used for preparation. After sheeting, pretreatment was carried out according to the method of the present invention (shown in Table 3). In this example, different treatment fluid formulations were tested (as shown in Table 3 below). Observation by bright field microscopy was performed after pretreatment. Then, FISH hybridization and DAPI staining were performed according to conventional methods, and observed under a fluorescent microscope.

[0065] table 3:

[0066]

[0067] The test results are shown in Table 3 and image 3 and Figure 4 As shown, among them, image 3 It shows the profile of the exfoliated cell droplet in normal human urine under bright field microscope after the solution of different formulations pretreated the exfoliated cells in urine at room temperature; Figure 4 It shows the FISH signal under the fluorescence microscope after the normal human urine ...

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 provides a pretreatment solution, a kit and a pretreatment method for fluorescence in situ hybridization of cell samples. Wherein, the pretreatment solution includes: 20-110 mM NaOH or KOH; and 0.1-1.0% non-ionic surfactant. By using the non-ionic surfactant in the above concentration range, the cell membrane can be destroyed to a certain extent but the basic shape and structure of the cell nucleus can be maintained. The lye in the above concentration range processes the cell sample, denatures the DNA and protein in the cell, and exposes the nucleic acid bound to the protein. In addition, non-ionic surfactants also help to increase the surface tension of cells and thus facilitate the loosening of the entire cell structure, which facilitates the efficient hybridization of DNA in the nucleus with fluorescent probes. The use of the pretreatment liquid has short treatment time, low cost and good stability of treatment effect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a pretreatment liquid, a kit and a pretreatment method for fluorescence in situ hybridization of cell samples. Background technique [0002] Fluorescence in situ hybridization (FISH) is a new technology developed in the 1980s that combines molecular biology and cytogenetics. Its principle is to use fluorescently labeled nucleic acid probes to hybridize with complementary regions of nucleic acids in samples. At present, it has been widely used in many fields such as genetic disease diagnosis, virus infection analysis, prenatal diagnosis, tumor genetics and genome research. [0003] Cytological samples include blood cells and various exfoliated cells, including urine, cervical samples, pleural fluid, amniotic fluid, etc., which contain exfoliated cells from various tissues and are widely used in clinical diagnosis and scientific research. For example, exfoliated cells in u...

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 Patents(China)
IPC IPC(8): C12Q1/6841C12Q1/6806
CPCC12Q1/6841C12Q1/6806C12Q2543/10C12Q2563/107C12Q2527/125C12Q2565/50
Inventor 丁凤王芳高梦娇凌萧洁李轶亢陈皓媛
Owner JIAXING ACCB DIAGNOSTICS
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