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Vacuum dyeing method for ultrathin sections of electron microscope

A technology of ultra-thin sectioning and staining, which is applied in the preparation of test samples, instruments, scanning probe technology, etc., to achieve the effect of clean slices, good staining effect and short staining time

Active Publication Date: 2016-12-07
江西金域医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] In order to solve the defects in the electron microscope ultra-thin section staining method in the prior art, the purpose of the present invention is to provide a vacuum staining method for the electron microscope ultra-thin section to solve the above defects

Method used

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  • Vacuum dyeing method for ultrathin sections of electron microscope
  • Vacuum dyeing method for ultrathin sections of electron microscope
  • Vacuum dyeing method for ultrathin sections of electron microscope

Examples

Experimental program
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Effect test

Embodiment 1

[0032] Embodiment 1, a kind of vacuum staining method of electron microscope ultrathin section

[0033] S1 Load 80 pieces of the copper nets with slices on the dyed silica gel plate according to the serial number to get the stained plate;

[0034] S2 places the stained plate prepared in step S1 in a petri dish, and puts the petri dish in a vacuum preservation box, then absorbs the supernatant of the uranyl acetate solution, which is prepared by mixing 4 g of acetic acid The uranyl powder is dissolved in 100ml of methanol with a volume concentration of 68% and stirred and dissolved; centrifuged in a centrifuge at a speed of 6000r / min for 12min, and the supernatant is taken; the supernatant is added to the staining plate, covered and cultivated The lid of the dish is vacuumized, and the vacuum degree is 30.41kpa, and it is dyed for 30min under vacuum conditions;

[0035] S3 recover the uranyl acetate solution dyed in step S2, put the stained plate on the shelf, then rinse with ...

Embodiment 2

[0037] Embodiment 2, a kind of vacuum staining method of electron microscope ultrathin section

[0038] S1 Load 90 pieces of the copper nets with slices on the dyed silica gel plate according to the serial number to obtain the dyed plate;

[0039] S2 places the stained plate prepared in step S1 in a petri dish, and puts the petri dish in a vacuum preservation box, then absorbs the supernatant of the uranyl acetate solution, which is prepared by mixing 4 g of acetic acid The uranyl powder is dissolved in 100ml of methanol with a volume concentration of 68% and stirred and dissolved; centrifuged in a centrifuge with a speed of 6000r / min for 14min, and the supernatant is taken; the supernatant is added to the staining plate, covered and cultivated The lid of the dish is vacuumized, and the vacuum degree is 34.31kpa, and it is dyed for 30min under vacuum conditions;

[0040] S3 recover the uranyl acetate solution dyed in step S2, put the stained plate on the shelf, then rinse wit...

Embodiment 3

[0042] Embodiment 3, a kind of vacuum staining method of electron microscope ultrathin section

[0043] S1 Load 100 pieces of copper nets with slices on the dyed silica gel plate according to the serial number to get the dyed plate;

[0044] S2 places the stained plate prepared in step S1 in a petri dish, and puts the petri dish in a vacuum preservation box, then absorbs the supernatant of the uranyl acetate solution, which is prepared by mixing 4 g of acetic acid The uranyl powder is dissolved in 100ml of methanol with a volume concentration of 68% and stirred and dissolved; centrifuged in a centrifuge with a speed of 6000r / min for 16min, and the supernatant is taken; the supernatant is added to the staining plate, covered and cultivated Dish cover, carry out vacuum treatment, described vacuum degree is 39.21kpa, dyeing 30min under vacuum condition;

[0045] S3 recover the uranyl acetate solution dyed in step S2, put the dyed plate on the shelf, then rinse with double distil...

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Abstract

The invention belongs to the technical field of sections of electron microscopes and particularly relates to a vacuum dyeing method for ultrathin sections of an electron microscope. The vacuum dyeing method for the ultrathin sections of the electron microscope comprises the following steps: copper wire meshes with sections fished are loaded to a dyeing silica gel plate in sequence, the dyeing silica gel plate is placed into a vacuumizing preservation box for dyeing with uranyl acetate, then dyeing with lead citrate is performed under a vacuum condition, and the ultrathin sections are obtained. According to the vacuum dyeing method for the ultrathin sections of the electron microscope, contact of CO2 in the air with lead citrate can be effectively isolated, and pollution to the sections and influence on electron microscopy observation due to a lead carbonate precipitate generated through a reaction of CO2 in the air and a lead salt are avoided. Meanwhile, the sections prepared with the vacuum dyeing method for the ultrathin sections of the electron microscope are clean, pollution-free, clear in structure, good in contrast effect and more beneficial to study of ultrastructures.

Description

technical field [0001] The invention belongs to the technical field of electron microscope sectioning, and in particular relates to a vacuum staining method for electron microscope ultrathin sections. Background technique [0002] For ultrathin sections of biological samples, the image contrast comes from the sample's ability to scatter the electron beam, which depends on the atomic composition. The higher the atomic number, the higher the electron density, the stronger the ability to scatter electrons, and it appears black under the electron microscope; the lower the atomic number, the lower the electron density, the weaker the ability to scatter electrons, and it appears white under the electron microscope. Biological samples are mainly composed of elements with low atomic numbers such as C, H, O, N, P, S, etc. Unstained ultra-thin sections have very weak contrast, especially for sections used for medical diagnosis. The structure is not clear, which seriously affects the ...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01Q30/20
CPCG01N1/30G01Q30/20
Inventor 侯晓涛王家健陶然王伶唐良玉冯强李改莫南勋陈建波汪垚
Owner 江西金域医学检验实验室有限公司
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