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Fixed liquid special for electron microscopy sample preparation of biological tissue sample and preparation method of fixed liquid

A technology of biological tissue and fixative, which is applied in the field of special fixative for electron microscope sample preparation of biological tissue samples and its preparation, which can solve the problems of poor preservation of microtubules and achieve the effect that bacteria are not easy to grow

Inactive Publication Date: 2018-11-30
北京艾普希隆生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Therefore, the present invention proposes a special fixative for electron microscope sample preparation of biological tissue samples and its preparation method, which is used to solve the problem of poor preservation of microtubules by common fixatives

Method used

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Effect test

preparation example Construction

[0019] A method for preparing a special fixative for electron microscope sample preparation of biological tissue samples, comprising the following steps:

[0020] Step 1: Dissolve 1.0-3.0g of paraformaldehyde powder in 20.0-30.0ml of distilled water, raise the temperature to below 65°C and keep stirring;

[0021] Step 2: Add 1.0-3.0ml of 1N sodium hydroxide while stirring. After the solution is transparent, add 3.2-6.6ml of 5% glutaraldehyde after cooling;

[0022] Step 3, add cacodylate buffer solution to the mixed solution to make the volume reach 50ml; the preparation method of cacodylate buffer solution is: prepare 4.0-5.2g sodium cacodylate, add distilled water to 100ml; 0.2N 1 - 20ml of hydrochloric acid, mix and dilute sodium cacodylate solution with 0.2N 1-20ml hydrochloric acid to 200ml to obtain cacodylate buffer solutions with different pH;

[0023] Step 4, finally add 22.0-26.2 mg of anhydrous calcium chloride, adjust the pH of the mixed solution to 6.8-7.5 to obt...

Embodiment 1

[0025] Dissolve 1.2g of paraformaldehyde powder in 20.0ml of distilled water, raise the temperature to below 65°C and keep stirring; add 1.0ml of 1N sodium hydroxide while stirring, after the solution is transparent, add 5% glutaraldehyde 3.2 after cooling ml; add cacodylic acid buffer solution to the mixed solution to make the volume reach 50.0ml;

[0026] The preparation method of cacodylate buffer solution is as follows: prepare 4.0g sodium cacodylate, add distilled water to 100ml; 0.2N1-20ml hydrochloric acid, mix and dilute sodium cacodylate solution with 0.2N 1-20ml hydrochloric acid to 200ml, Cacodylate buffer solution with different pH was obtained, see Table 1 for details;

[0027] pH

6.8

7.0

7.2

7.4

7.6

7.8

Sodium cacodylate ml

50

50

50

50

50

50

0.2N hydrochloric acidml

18.3

13.3

9.3

6.3

4.2

2.7

Add double distilled water to ml

200

200

200

200

200

200

[0028] ...

Embodiment 2

[0031] Dissolve 2.0g of paraformaldehyde powder in 25.0ml of distilled water, raise the temperature to below 65°C and keep stirring; while stirring, add 3.0ml of 1N sodium hydroxide, after the solution is transparent, cool and add 5% glutaraldehyde 5.0 ml; add cacodylate buffer solution to the mixed solution to make the volume reach 50.0ml; the preparation method of cacodylate buffer solution is: prepare 4.0g sodium cacodylate, add distilled water to 100ml; 0.2N 1-20ml For hydrochloric acid, mix sodium cacodylate solution with 0.2N 1-20ml hydrochloric acid and dilute it to 200ml to obtain cacodylate buffer solutions with different pHs, see Table 2 for details;

[0032] pH

6.4

6.6

6.8

7.0

7.2

7.4

Sodium cacodylate ml

50

50

50

50

50

50

0.2N hydrochloric acidml

18.3

13.3

9.3

6.3

4.2

2.7

Add double distilled water to ml

200

200

200

200

200

200

[0033] Table II

[0034] Step...

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Abstract

The invention relates to the technical field of preparation of an electron microscopy sample preparation fixed liquid, in particular to a fixed liquid special for electron microscopy sample preparation of biological tissue sample. The fixed liquid comprises the following constituents: 1.3-3.0g paraformaldehyde powder, 20.0-30.0ml distilled water, 1.0-5.0ml 1N sodium hydroxide, 3.0-6.0ml 5% glutaraldehyde, a dimethylarsinic acid buffer liquid and 2.20-26.0mg anhydrous calcium chloride. The preparation method of the fixed liquid comprises the following steps of 1, taking and dissolving 1.3-3.0gparaformaldehyde powder in the 20.0-30.0 distilled water, rising a temperature to 65 DEG C or below and constantly stirring; 2, simultaneously adding 1.0-3.0ml 1N sodium hydroxide during stirring, adding 3.0-6.0ml 5% glutaraldehyde after cooling and after the solution is transparent; 3, adding the dimethylarsinic acid or phosphate buffer liquid into a mixed solution so that volume reaches 50ml; and 4, adding 22.0-26.2ml anhydrous calcium chloride, and adjusting pH of the mixed liquid to 6.8-7.5, thereby obtaining the fixed liquid of the product.

Description

technical field [0001] The invention relates to the technical field of preparation of a fixative for electron microscope sample preparation, in particular to a special fixative for electron microscope sample preparation of biological tissue samples and a preparation method thereof. Background technique [0002] With the development of precision medicine, observing and understanding the ultra-microscopic structure of various organelles in a single cell under physiological and pathological conditions is of great significance for basic medical research and clinical pathological diagnosis. At present, light microscope and electron microscope are indispensable tools for the structural observation of animal cells. The structure image displayed by light microscope observation is limited by its magnification, and the structural information is very limited; the transmission electron microscope can observe the ultrastructure of cells ( Sui Shuguang.Experiment of using electron microsc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
CPCG01N1/28
Inventor 晋玉宽
Owner 北京艾普希隆生物科技有限公司
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