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Immumofluorescent antibody labeling method of cotton tender tissue microtube framework

An immunofluorescence and antibody labeling technology, which is applied in the field of cotton scientific research, can solve the problems of no fluorescent antibody labeling, etc., and achieve the effects of reducing the formation of ice crystals or larger ice crystals, reducing cell damage, and preserving the activity of biomolecules

Inactive Publication Date: 2017-05-31
INST OF IND CROPS HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, fluorescent antibodies have not yet been labeled on the tissue level of cotton, such as the microtubule skeleton at the shoot tip and the microtubule skeleton at the root tip of cotton.

Method used

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  • Immumofluorescent antibody labeling method of cotton tender tissue microtube framework
  • Immumofluorescent antibody labeling method of cotton tender tissue microtube framework
  • Immumofluorescent antibody labeling method of cotton tender tissue microtube framework

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] In this example, immunofluorescent labeling was performed on the microtubule skeleton of the outer cortex cells of the main stem 1 cm below the shoot tip of the cotton three-leaf stage.

[0047] A method for immunofluorescence antibody labeling of cotton young tissue microtubule skeleton, comprising the following steps:

[0048] The first step, fixative fixation

[0049]Cut the outer skin of the main stem 1 cm below the tip of the cotton three-leaf stage with bare hands, put the outer skin of the main stem into a sealed container filled with fixative, vacuumize the sealed container for 15 minutes, and then place the sealed container at 28°C For 3 hours, the fixative in the sealed container fixes the young cotton tissue;

[0050] Described stationary liquid is to contain the glutaraldehyde that mass percent is 0.4%, the paraformaldehyde that mass percent is 3.6%, the Triton-100 that mass percent is 0.02%, pipes, MEGTA and MgSO , wherein the molar concentration of pipes ...

Embodiment 2

[0068] Repeat Example 1, with the following differences. In this example, the root cortex at 1 cm above the root tip of the cotton three-leaf stage is used for immunofluorescent antibody labeling. The specific implementation steps are as follows:

[0069] The first step, fixative fixation

[0070] Cut the root cortex at 1 cm above the root tip of cotton at the three-leaf stage with bare hands, put the root cortex into a sealed container filled with fixative, vacuumize the sealed container for 10 minutes, and then put the sealed container at 28 ° C for 2 Hours, the fixative in the airtight container fixes the tender tissue of cotton;

[0071] Described stationary liquid is to contain the glutaraldehyde that mass percent is 0.4%, the paraformaldehyde that mass percent is 3.6%, the Triton-100 that mass percent is 0.02%, pipes, MEGTA and MgSO , wherein the molar concentration of pipes is 50 mM, the molar concentration of MEGTA is 5 mM, and the molar concentration of MgSO4 is 5 mM...

Embodiment 3

[0089] Example 1 was repeated, with the following differences. In this example, immunofluorescent labeling was performed on the microtubule skeleton of the root epidermis at 0.5 cm above the root tip of cotton at the three-leaf stage.

[0090] A method for immunofluorescence antibody labeling of cotton young tissue microtubule skeleton, comprising the following steps:

[0091] The first step, fixative fixation

[0092] Cut the root cortex at 0.5 cm above the root tip of cotton at the three-leaf stage with bare hands, put the root cortex into a sealed container filled with fixative, vacuumize the sealed container for 8 minutes, and then place the sealed container at 30°C for 2 Hours, the fixative in the airtight container fixes the tender tissue of cotton;

[0093] Described stationary liquid is to contain the glutaraldehyde that mass percent is 0.4%, the paraformaldehyde that mass percent is 3.6%, the Triton-100 that mass percent is 0.02%, pipes, MEGTA and MgSO , wherein the ...

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Abstract

The invention provides an immumofluorescent antibody labeling method of a cotton tender tissue microtube framework. The method comprises the following process steps: performing fixation by a fixing liquid; performing dehydration; freezing a microtome section; removing non-specific fluorescence; and performing immunolabelling; performing section sealing and the like. The method provided by the invention combines fluorescein with the microtube framework at the cotton tender tissue to immunolabel the cotton tender tissue microtube framework so as to observe changes of the cotton tender tissue microtube framework in the tissue level and research resistance of cottons to non-biological adversity stress and resistance of cottons on plant diseases and insect pests. The method provided by the invention is simple in step, easy to master, clear in result and suitable for observing the cotton tender tissue microtube framework, and also has certain reference value in observing other plant tender tissue microtube frameworks.

Description

[0001] Technical field: [0002] The invention belongs to the technical field of cotton scientific research, and in particular relates to an immunofluorescent antibody labeling method for the microtubule skeleton of young cotton tissues. [0003] Background technique: [0004] Cotton is one of the important economic crops in my country. Some abiotic stresses such as low temperature, drought and salt damage seriously affect the growth and development of cotton. At the same time, pests and diseases are also one of the important reasons affecting the growth and development of cotton. Stress resistance and cotton's resistance to diseases and insect pests are important contents of scientific research on cotton by researchers. [0005] Fluorescent antibody labeling technology is to chemically combine fluorescein with specific antibodies to form a fluorescein-protein conjugate, that is, a fluorescently labeled antibody. This conjugate still retains the activity of the antibody and has ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533
CPCG01N33/533
Inventor 赵付安李艺李武房卫平侯甲男
Owner INST OF IND CROPS HENAN ACAD OF AGRI SCI
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