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A kind of pipetting solution for frozen ultrathin section of polymer material and preparation method of section

A technology for freezing ultra-thin sections and polymer materials, applied in sampling devices and other directions, can solve the problems of inability to distinguish different structures and forms, complicated steps, introduction of chemical reagents, etc., and achieves a simple and easy preparation method, simple composition, and operation steps. simple effect

Active Publication Date: 2018-04-10
CHINA PETROLEUM & CHEM CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The wet knife method is similar to ultra-thin sectioning at room temperature. The dimethyl sulfoxide solution that does not freeze at low temperature is used as the tank body, and the section is unfolded after floating on the solution, and the sample is taken with a pre-cooled filmed copper grid; the method’s The disadvantage is that dimethyl sulfoxide solution has certain toxicity, and this method is not applicable at extremely low temperature, and has been gradually replaced by dry knife method
However, when sucrose phosphate is used as the pipetting solution, the pipetting solution on the section needs to be washed before staining, and when observed under the electron microscope, a small amount of residual pipetting solution is easily carbonized under electron beam bombardment, resulting in sample Decreased resolution or inability to distinguish between different structural morphologies
In addition, dyes such as osmic acid can undergo oxidation-reduction reactions with aldehyde groups in sugars, reducing the contrast of the sample and reducing the clarity of the image.
[0008] In general, the above methods have certain defects for the removal of sections in frozen ultrathin sections of polymer samples, such as limitations by temperature and sample preparation tools, complicated steps, introduction of new chemical reagents or toxicity, reduced Image resolution and clarity, etc.

Method used

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  • A kind of pipetting solution for frozen ultrathin section of polymer material and preparation method of section
  • A kind of pipetting solution for frozen ultrathin section of polymer material and preparation method of section
  • A kind of pipetting solution for frozen ultrathin section of polymer material and preparation method of section

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Embodiment 1

[0031]Add deionized water, absolute ethanol and cyclohexanol (deionized water: absolute ethanol: cyclohexanol = 1: 1.5: 0.5) drop by volume on a clean glass slide to prepare frozen ultrathin section pipetting solution , and then immerse the platinum ring in the pipetting solution. Put the platinum ring dipped in the pipetting solution into the freezer of the microtome, freeze at -140°C for 20s, and then stick the cut styrene-butadiene-isoprene ternary under the blade of the slicer Copolymer (SIBR) sliced ​​samples. Remove the platinum ring with the sliced ​​sample and place it horizontally on the copper grid. The side of the platinum ring with the sliced ​​sample faces down and is in contact with the copper grid. Use filter paper to suck the pipetting liquid from under the copper grid and remove it after 1 min. Platinum ring, so that the ultra-thin section sample is attached to the upper surface of the copper grid. After the copper grid was dried, the sections were stained w...

Embodiment 2

[0035] Add deionized water, absolute ethanol and pentaerythritol (deionized water: absolute ethanol: pentaerythritol = 4: 1: 1) drop by volume on a clean glass slide to prepare frozen ultrathin section pipetting solution, and then transfer the platinum The ring is immersed in the pipetting solution. Put the platinum ring dipped in the pipetting solution into the freezer chamber of the microtome, freeze at -110°C for 10 seconds, and then stick the cut styrene-butadiene-acrylonitrile terpolymer under the blade of the slicer (ABS) sliced ​​samples. Remove the platinum ring with the sliced ​​sample and place it horizontally on the copper grid. The side of the platinum ring with the sliced ​​sample faces down and is in contact with the copper grid. After 2 minutes, remove the platinum ring to make the ultrathin sliced ​​sample stick flat. Attached to the upper surface of the copper mesh. After the copper grid was dried, the sections were stained with osmic acid for 3 hours, and t...

Embodiment 3

[0039] Add deionized water, absolute ethanol and glycerol (deionized water: absolute ethanol: glycerol = 1: 0.5: 1.5) drop by volume on a clean glass slide to prepare frozen ultrathin section pipetting solution , and then immerse the platinum ring in the pipetting solution. Put the platinum ring dipped in the pipetting solution into the freezer of the microtome, freeze at -30°C for 10s, and then stick the cut polypropylene / SiO 2 Sliced ​​samples. Remove the platinum ring with the sliced ​​sample and place it horizontally on the copper grid. The side of the platinum ring with the sliced ​​sample faces down and is in contact with the copper grid. After 2 minutes, remove the platinum ring to make the ultrathin sliced ​​sample stick flat. Attached to the upper surface of the copper mesh. After the copper grid was dried, the sections were directly observed with a transmission electron microscope, and the results were as follows: image 3 As shown in A.

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Abstract

The invention provides a pipetting solution for frozen ultrathin sections of polymer materials, which is composed of water and organic alcohol compounds with a volume ratio of 6:1-1:8. The composition is simple, the preparation method is simple and easy, there is no need to wash the slices after use, and the ratio can be easily adjusted according to the freezing temperature of the sliced ​​materials, so as to achieve the best pipetting effect. The present invention also provides a method for preparing frozen ultra-thin sections of macromolecular materials by using the above-mentioned pipetting liquid to pipette sections. The operation steps are simple, suitable for the use of glass knife or diamond knife, and there is no damage to the knife edge. After the section is removed, the section does not need to be rinsed, and can be directly stained, thereby reducing the operation steps. The invention provides a high-efficiency and simple method for preparing frozen ultra-thin sections of polymer materials for transmission electron microscope testing.

Description

technical field [0001] The invention relates to a pipetting liquid for frozen ultrathin sections of polymer materials and a preparation method for frozen ultrathin sections of polymer materials using the pipetting liquid. Background technique [0002] Transmission Electron Microscope is one of the important instruments for studying the microstructure of polymer materials. However, the premise of being able to observe the microstructure of materials on a transmission electron microscope is that the electron beam of the electron microscope can penetrate the material sample, and the quality of the observed image strongly depends on the thickness of the sample. Therefore, the electron microscope sample must be very thin, and the general thickness needs to be controlled between 10-100nm. [0003] Ultrathin sectioning is the most basic and important sample preparation technique in polymer sample preparation methods for transmission electron microscopy. However, for polymer mater...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/06
Inventor 杨娟刘滢郭曦
Owner CHINA PETROLEUM & CHEM CORP