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Transmission electron microscope embedding pretreatment method for liquid materials

A technology for transmission electron microscopy and liquid samples, which is applied in the fields of analyzing materials, using wave/particle radiation for material analysis, measuring devices, etc.

Inactive Publication Date: 2019-05-03
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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  • Transmission electron microscope embedding pretreatment method for liquid materials
  • Transmission electron microscope embedding pretreatment method for liquid materials
  • Transmission electron microscope embedding pretreatment method for liquid materials

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 transmission electron microscope embedding pretreatment method for liquid materials. The method includes the following steps: (1) sealing one end of a capillary burette, and immersing the other end of the capillary burette into melted agarose gel so that the surface on one end of the capillary burette can be uniformly stuck with the agarose gel to form the tubular agarosegel, and placing the capillary burette stuck with the agarose gel on freezing metal for 1-1.5 min; (2) placing slide glass on the freezing metal, and separating the tubular agarose gel obtained by thestep (1) from the capillary burette to put on the slide glass; (3) cutting the agarose gel into a small tube, and adding a liquid sample into the small tube; and (4) sealing the openings on the two ends of the small tube by using the agarose gel, and after the agarose gel is completely solidified, cutting the agarose gel having the small tube, and performing fixing with aldehyde. The method cannot change appearance, so that the real appearance of substances can be actually reflected; and the method has no lowest requirements for the concentration of the liquid sample.

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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IPC IPC(8): G01N23/2202G01N23/2251
Inventor 贺延玉杜平刘秀
Owner GANSU AGRI UNIV