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Pear fruit stone cell specific histochemical localization method

A technique of histochemistry and pear fruit, applied in the field of cell staining, can solve the problems of accelerated browning of pear pulp, burnt skin, corroded skin, etc., and achieves the effects of high discrimination, sensitive response and easy operation.

Active Publication Date: 2020-10-30
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] current routine The Wiesner method only stains the lignin, which will stain the lignin in the stone cells, peel, pulp, fruit core, fruit stalk and other parts, causing background interference
This dyeing method is not only easy to fade and high cost, but also the reagents used such as: concentrated hydrochloric acid, concentrated ammonia water, potassium permanganate, etc. are all hazardous chemicals. Once inhaled, it will cause serious damage to the respiratory tract and other parts. If you accidentally touch it, you will burn skin, corrodes the skin, and strong acid and alkali will also accelerate the browning of pear pulp

Method used

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  • Pear fruit stone cell specific histochemical localization method
  • Pear fruit stone cell specific histochemical localization method
  • Pear fruit stone cell specific histochemical localization method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Diaminobenzidine-specific histochemical localization of stone cells

[0044] The pear fruit is placed in the vitamin C solution of 0.05g / L and sliced ​​(the thickness of the section is 3cm), then the pear fruit slice is submerged in the diaminobenzidine solution with a concentration of 0.5g / L, and transferred to the light incubator Soak in dark and culture for 4 hours (reagents are easy to decompose when exposed to light), adjust the temperature to 24°C, decolorize and rinse with decolorizing agent, seal the slide with FAA fixative solution and observe under ordinary optical microscope. The staining results are as follows: figure 1 shown. Wherein the decolorizing agent is glycerin solution and ethanol solution, the volume ratio of glycerol solution and ethanol solution is 1:1, the volume percentage content of glycerin solution is 30%, and the volume percentage content of ethanol solution is 70%.

[0045] from figure 1 It can be seen that all the stone cells are dyed r...

Embodiment 2

[0047] Nitrotetrazolium-stone cell-specific histochemical localization

[0048] Pear fruit is placed in 0.05g / L vitamin C solution and sliced ​​(the thickness of the slice is 3cm), then the pear fruit slice is submerged in the nitrotetrazole solution with a concentration of 0.5g / L, and transferred to light culture Soak in the box and cultivate in the dark for 4 hours (the reagent is easy to decompose when exposed to light), the temperature is adjusted to 24°C, the decolorizing agent is decolorized and rinsed, and the slide is sealed with FAA fixative solution for observation. The staining results are as follows: figure 2shown. Wherein the decolorizing agent is glycerin solution and ethanol solution, the volume ratio of glycerol solution and ethanol solution is 1:1, the volume percentage content of glycerin solution is 30%, and the volume percentage content of ethanol solution is 70%.

[0049] from figure 2 It can be seen that the stone cells are stained blue-gray, and the ...

Embodiment 3

[0051] Comparison of specific histochemical localization of pear stone cells in Dangshansu pear and Liangli Zaosu

[0052] Put the Dangshansu pear fruit and Liangli Zaosu fruit in 0.05g / L vitamin C solution to slice (3cm), and then submerge the pear fruit slices in 0.5g / L diaminobenzidine solution and nitric acid solution respectively. In the tetrazolium-based solution, transfer to the light incubator and soak in the dark for 4 hours (the reagent is easy to decompose when exposed to light), the temperature is adjusted to 24°C, the decolorizing agent is decolorized and rinsed, and the slide is sealed with FAA fixative solution for observation. The staining results are as follows: Figure 3-2 with Figure 3-3 shown. Wherein the decolorizing agent is glycerin solution and ethanol solution, the volume ratio of glycerol solution and ethanol solution is 1:1, the volume percentage content of glycerin solution is 30%, and the volume percentage content of ethanol solution is 70%.

[...

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Abstract

The invention provides a pear fruit stone cell specific histochemical localization method, and belongs to the technical field of cell staining. The method comprises the following steps: slicing pear fruits in a vitamin C solution to obtain pear fruit slices; submerging the pear fruit slices in a dyeing reaction solution for 2-5 hours to obtain soaked pear fruit slices, wherein the components of the dyeing reaction solution comprise diaminobenzidine or nitrotetrazole; and decolorizing the soaked pear fruit slices by using a decolorizing agent, and sealing the slices by using a stationary liquid. According to the invention, the method is high in distinguishing degree, and can accurately, conveniently and effectively distinguish the pear pulp parenchyma cells from the stone cells; the safetycoefficient is high, hazardous chemical substances are not involved, and pungent smell is avoided; vitamin C effectively reduces the browning phenomenon of slices and prevents color development colorinterference; and morphological structure characteristics in the stone cell development process can be efficiently observed, and pear varieties with different stone cell contents and sizes can be distinguished.

Description

technical field [0001] The invention belongs to the technical field of cell staining, in particular to a method for specific histochemical localization of stone cells in pear fruit. Background technique [0002] Pear is one of the important deciduous fruit trees in my country, with a long history of cultivation, wide distribution and abundant resources. Stone cells are formed by the deposition of lignin on the cell wall of parenchyma cells in pear fruit, which causes secondary thickening of the cell wall. The content of pear stone cells is a decisive factor for the quality of pear fruit. Therefore, it is of great significance to study the morphology, distribution and tissue localization of pear stone cells. [0003] Previous studies have shown that the formation of pear stone cells is a process of programmed cell death, and the period of cell apoptosis is closely related to the accumulation of reactive oxygen species, which play a key role in the polymerization and depositi...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N1/31G01N21/84
CPCG01N1/30G01N1/31G01N21/84G01N2021/8466
Inventor 蔡永萍章扬程曦金青黄满畅鲁津津
Owner ANHUI AGRICULTURAL UNIVERSITY
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