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Pre-treatment method of liquid material for TEM embedding

A technology of transmission electron microscopy and liquid samples, which is applied in the direction of analyzing materials, using wave/particle radiation for material analysis, instruments, etc., and can solve problems such as inconvenient observation, no way to embed, and sample dispersion

Inactive Publication Date: 2021-06-04
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Centrifugation will cause changes in the shape of the tangible substance of the sample, which cannot truly reflect the true shape of the substance.
And when the sample concentration is very low, the sediment sample cannot be obtained after centrifugation, and there is basically no way to embed it.
Moreover, when the traditional method is used to embed the obtained precipitate, it is easy to disperse the sample in the agarose gel, which causes a lot of inconvenience to the observation after sectioning.

Method used

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  • Pre-treatment method of liquid material for TEM embedding
  • Pre-treatment method of liquid material for TEM embedding
  • Pre-treatment method of liquid material for TEM embedding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The steps of the transmission electron microscope embedding method of the liquid sample of the present invention are as follows:

[0027] 1. Make fresh 2.5%-3.5% agarose gel (0.5g-0.7g agarose dissolved in 20ml of water), let it fully dissolve between 80-90°C. Then place it in a 55-65°C water bath for later use.

[0028] 2. Freeze a piece of metal in the refrigerator in advance, it doesn’t need to be too big, just enough to fit a glass slide. There is no requirement for the material of the metal, as long as the metal surface is flat.

[0029] The purpose of the frozen metal is to quickly allow the gel to cool and set.

[0030] 3. Choose a tapered capillary pipette (see figure 1 ), the inner diameter of one end of the gel tube is 1 mm, and the other end is 5 mm. The thick end of the capillary dropper is sealed with the index finger, and the thin end of the capillary dropper is immersed in the agarose gel in the water bath for about 10 -20 seconds, the immersion depth...

Embodiment 2

[0043] Embodiment 2 application embodiment

[0044] The research on the digestion process of milk has always been a research hotspot at home and abroad. How to witness the digestion process through transmission electron microscopy has always been a difficult problem to overcome. Even through centrifugation, it is difficult to separate the formed components, and the formed components are delaminated and deformed. However, the method of the present invention can well solve this problem and is applicable to all milk sample materials.

[0045] Centrifuge the formed components of milk and embed them in agarose gel. figure 2 .

[0046] After the milk is processed and embedded by the method in this embodiment 1, the results of transmission electron microscope observation are shown in image 3 and Figure 4 .

[0047] figure 2 After centrifuging the formed components of milk, they were embedded in agarose gel and observed in ultra-thin sections, the cells were seriously deform...

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Abstract

The invention provides a method for pre-embedding a liquid sample in a transmission electron microscope, comprising the following steps: (1), sealing one end of the capillary dropper, immersing the other end of the capillary dropper in melted agarose gel, making one end of the capillary dropper Evenly stick the agarose gel on the surface to form a tubular agarose gel, then place the capillary pipette with the agarose gel on the frozen metal for 1-1.5 minutes; (2), place the slide glass on the frozen metal slice, the tubular agarose gel obtained in step (1) is separated from the capillary dropper onto the glass slide; (3), the agarose gel is cut into small tubes, and the liquid sample is added to the small tube; (4), with The agarose gel seals the openings at both ends of the tubules, and after the agarose gel is completely solidified, the agarose gel with the tubules is cut and fixed with aldehyde. The method of the invention does not cause changes in appearance, can truly reflect the real appearance of substances, and has no minimum requirement on the concentration of liquid samples.

Description

technical field [0001] The invention relates to a method for pre-embedding a liquid material in a transmission electron microscope. Background technique [0002] The traditional TEM embedding method requires a high concentration of liquid material, and the method of centrifuging to collect sediment is basically used for embedding. Centrifugation will cause changes in the shape of the tangible substance of the sample, which cannot truly reflect the true shape of the substance. And when the sample concentration is very low, no sediment sample can be obtained after centrifugation, and there is basically no way to embed it. Moreover, when the obtained precipitate is embedded in the traditional method, the sample is easily dispersed in the agarose gel, which causes a lot of inconvenience to the observation after sectioning. Contents of the invention [0003] In order to solve the problems existing in the prior art, the present invention provides a pre-treatment method for TEM...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N23/2202G01N23/2251
Inventor 贺延玉杜平刘秀
Owner GANSU AGRI UNIV