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Endosperm tissue embedding method and application thereof

An endosperm and resin technology, applied in the field of plant tissue embedding, can solve problems such as difficulty in obtaining a fixed embedding effect

Active Publication Date: 2021-04-16
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the tight arrangement of rice endosperm starch granules, it is difficult to obtain a good fixed embedding effect by the conventional plant high-pressure freezing-freezing substitution method

Method used

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  • Endosperm tissue embedding method and application thereof
  • Endosperm tissue embedding method and application thereof
  • Endosperm tissue embedding method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Example 1 Fixation and embedding of rice endosperm samples 3 days after flowering

[0040] The present embodiment provides a method for embedding endosperm tissue of the present invention, specifically as follows:

[0041] 1. High pressure freeze fixation, take Leica EM PACT2 as an example.

[0042] 1) Preparation: Open the Leica EM PACT2, add a sufficient amount of liquid nitrogen to the liquid nitrogen chamber, add the pressure liquid methylcycloethane, and use the empty carrier (carrier, Flat Specimen Carrier, Leica, cat#16706899) ​​to high-pressure freeze three times , make the machine in a stable state, and confirm whether the temperature and pressure curves are normal.

[0043] 2) Sample preparation: Take the rice endosperm 3 days after fresh flowering and put it in 0.15M sucrose solution, use a single-sided blade to cross-cut the rice endosperm, take the endosperm tissue with a thickness of 400 μm and a diameter of 1.2 mm, and put it into a carrier with a thickn...

Embodiment 2

[0057] Example 2 Fixation and embedding of rice endosperm samples 12 days after flowering

[0058] This embodiment provides a method for embedding endosperm tissue of the present invention, specifically as follows (parts not described in detail are the same as in Example 1):

[0059] 1. High pressure freeze fixation, take Leica EM PACT2 as an example.

[0060] 1) Preparation: Open the Leica EM PACT2, add a sufficient amount of liquid nitrogen to the liquid nitrogen chamber, add the pressure liquid methylcycloethane, and freeze it three times with an empty carrier under high pressure to keep the machine in a stable state, and confirm whether the temperature and pressure curves are correct. normal.

[0061] 2) Sample preparation: Take fresh rice endosperm 12 days after flowering and place it in 0.15M sucrose solution, use a single-sided blade to cross-cut the rice endosperm, take endosperm tissue with a thickness of 400 μm and a diameter of 1.2 mm, and put it into a carrier wit...

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Abstract

The invention relates to the technical field of plant tissue embedding, and particularly discloses an endosperm tissue embedding method and application thereof. The endosperm tissue embedding method disclosed by the invention is realized through a high-pressure freezing technology and a freezing substitution technology, and procedures, endosperm permeation and other links during freezing substitution embedding are explored, so that the obtained method can be used for obtaining a rice endosperm tissue sample capable of keeping the original cell structure form of a plant; and the method is suitable for rice endosperm samples and similar plant tissues in different development stages. The tissue sample embedded by the method can subsequently obtain high-quality ultrathin sections, can be used for observing the ultrastructure of a transmission electron microscope, and can also be used for carrying out positioning and quantitative research on protein under the ultrastructure through immune colloidal gold staining.

Description

technical field [0001] The invention relates to the technical field of plant tissue embedding, in particular to an endosperm tissue embedding method and application. Background technique [0002] The development of conventional electron microscope technology in life sciences makes it possible to observe the ultrastructure of tissues and cells. Although the resolution of transmission electron microscopy has reached the atomic level, in the process of biological sample preparation, chemical reagents such as formaldehyde and glutaraldehyde often have unpredictable effects on the chemical fixation of samples, which cannot fully reflect the true nature of tissues and cells. Morphological structure, and the antigenic preservation effect on macromolecular substances such as proteins is poor. High-pressure cryofixation technology can completely fix tissues and cells in a very short time without using any chemical fixatives, which can be combined with cryosubstitution to unify the f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
Inventor 张丽娜任玉龙潘天魏潜吴佳楠
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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