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A biomaterial dyeing method utilizing PIP

A technology of biological materials and dyeing methods, applied in the direction of analysis materials, test sample preparation, sampling, etc., can solve problems such as difficult to buy reagents

Active Publication Date: 2017-10-24
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to its radioactivity, reagent management and waste liquid treatment have always been issues that cannot be underestimated. In addition, the reagent is currently under strict control, and it is difficult for most scientific research institutions to buy this reagent except for the original inventory.

Method used

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  • A biomaterial dyeing method utilizing PIP
  • A biomaterial dyeing method utilizing PIP
  • A biomaterial dyeing method utilizing PIP

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, the application of PIP in negative staining

[0044] 1. Phage

[0045] Use PIP to negatively stain the phage, and use unstained and uranyl acetate to negatively stain the phage as a control. The specific steps for negatively staining the phage with PIP are as follows:

[0046] After the 200-mesh copper grid glow discharge hydrophilic treatment, take the phage suspension to be tested and drop it on the copper grid. After 1 minute, use filter paper to absorb the excess droplet, wash it with water once, and use 0.5% (mass percentage) PIP aqueous solution After staining for 1 minute, absorb excess liquid with filter paper, air-dry and observe under the microscope.

[0047] The procedure for negative staining of phage with uranyl acetate is as follows:

[0048] After the 200-mesh copper grid glow discharge hydrophilic treatment, take the phage suspension to be tested and drop it on the copper grid. After 1 minute, use filter paper to absorb the excess droplet...

Embodiment 2

[0063] Embodiment 2, the application of PIP in positive staining

[0064] Take plant leaf slices as an example to observe the role of PIP in positive staining.

[0065] Use PIP-lead citrate to double-stain the slices of the plant leaves, and use uranyl acetate-lead citrate to double-stain the slices of the plant leaves and lead citrate to single-stain the slices of the plant leaves as a control. Unstained plant leaf sections were used as unstained controls.

[0066] The operation steps of PIP-lead citrate double-staining the slices of plant leaves are as follows:

[0067] Spread a clean nitrocellulose membrane, drop 1% (mass percentage) PIP aqueous solution on the membrane, buckle the copper grid with slices on the droplet in the direction that the slice faces the PIP aqueous solution, and incubate for 25 minutes. After staining, immerse the net with tweezers in distilled water and wash repeatedly, then blot the excess liquid with filter paper. Then buckle the side containi...

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Abstract

A biomaterial dyeing method utilizing PIP is disclosed. The method includes dyeing a biomaterial by utilizing the PIP or a PIP-containing agent to complete biomaterial dyeing. The chemical structure of the PIP is shown as a formula I. The biomaterial can be microbes such as bacteriophages, viruses and bacteria, and can also be animal and plant cells, tissues, organs and sections thereof. Experiments prove that the PIP can be used for dyeing a plurality of biomaterials, and the PIP is not a dangerous compound, does not have physical or chemical danger, is harmless to health and environment, is easily available and water-soluble, and can replace uranyl acetate and be used for biomaterial dyeing.

Description

technical field [0001] The invention relates to a method for dyeing biological materials by using PIP in the field of biotechnology. Background technique [0002] Transmission electron microscopy is used to image the transmitted electrons through the sample. Most biological samples are composed of atoms with low atomic numbers such as carbon, hydrogen, and oxygen. In order to enhance the contrast of the sample, heavy metal dyeing is used to increase the scattering generated after collision with electrons. way to increase the contrast of the sample. According to the different ways in which heavy metals play an important role in imaging, the staining methods can be divided into negative staining and positive staining. Negative staining is mostly used for the observation of complete small particles, using heavy metal salts with higher electron density to surround the sample, so that the sample with low electron density can be contrasted against the electron-dense gray-black ba...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
CPCG01N1/30G01N2001/302
Inventor 梁静南刘一苇赵彤孙树涛樊峥李春立张晓兰王前
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI