Preparation method of frozen slices of traditional Chinese medicine cinnamomum cassia

A production method and cryosectioning technology, which are applied in the preparation, sampling, and measuring devices of test samples, can solve the problems of easy breaking, different cell structure, uneven cross-section, etc. short cycle effect

Pending Publication Date: 2021-05-04
鉴甄检测技术(上海)有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Disclosed in the Chinese invention patent (CN 103776657 A) is a kind of manufacturing method of coconut leaf frozen section, promptly adopts the frozen section mode of frozen section to obtain the frozen section of complete structure fast, but because: 1) leaf quality and cell tissue structure and bark Completely different; 2) and the sample implemented in this patent is a fresh leaf, while the traditional Chinese medicine cinnamon is a dry bark with a thickness of 0.2 to 0.8 cm. The outer surface is slightly rough, with irregular fine wrinkles and laterally protruding lenticels; the inner surface is slightly Flat, with fine longitudinal grains, hard and brittle, easy to break, and uneven cross-section; and the structure between cells and tissues is in an extremely dry state, and it cannot be cut without proper pretreatment methods
Therefore, the method for making frozen slices of leaves provided by this invention patent is not suitable for the frozen slices of the traditional Chinese medicine cinnamon

Method used

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  • Preparation method of frozen slices of traditional Chinese medicine cinnamomum cassia
  • Preparation method of frozen slices of traditional Chinese medicine cinnamomum cassia

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Effect test

Embodiment 1

[0049] The present embodiment provides a kind of preparation method of Chinese medicine cinnamon frozen section, comprises the following steps:

[0050] (1) Material pretreatment:

[0051] Take the traditional Chinese medicine cinnamon, use a tool to trim a section of 1 × 1cm sample; put it into a beaker filled with 70% ethanol: water = 1:1 (volume ratio), soak for 1 hour, take it out, and cut it into a length of 8mm with a blade. Take a short sample with a width of about 5mm, and smooth the edges; soak for another 2 hours to make the short sample moist, take it out, dry the surface solution, put it into a test tube containing chloral hydrate test solution, and heat to boiling for 6 times; take out the sample and dry the surface solution;

[0052] (2) Sample fixation:

[0053] Add OCT cryo-embedding agent to the base of the cold stage of the Leica CM1950 cryostat to meet the size of the short-section sample, pre-freeze for 20 seconds, take out the base, place the short-secti...

Embodiment 2

[0063] In this embodiment, the proportion of ethanol in the soaking solution in the preparation method is explored, and the specific operation method and experimental results are as follows:

[0064] (1) Material pretreatment:

[0065] Put the sample into a beaker filled with 70% ethanol:water=2:1 (volume ratio) solution, and the others are the same as in Example 1.

[0066] (2) Sample fixation: same as in Example 1.

[0067] (3) Frozen section: same as embodiment 1.

[0068] (4) film exhibition and film production: with embodiment 1.

[0069] (5) Microscopic observation and photographing: same as in Example 1.

[0070] Experimental results: In this method, the proportion of ethanol in the soaking solution is relatively large, although the time for soaking the sample is slightly shortened, but the water content of the obtained sample is low, and the intercellular combination is not tight enough. When other conditions remain unchanged, The resulting slices are more prone to...

Embodiment 3

[0072] This embodiment explores the soaking time in the pretreatment step in the preparation method, and the specific operation method and experimental results are as follows:

[0073] (1) Material pretreatment:

[0074] The pre-soaked trimmed sample was then soaked for 3.5 hours, and the others were the same as in Example 1.

[0075] (2) Sample fixation: same as in Example 1.

[0076] (3) Frozen section: same as embodiment 1.

[0077] (4) film exhibition and film production: with embodiment 1.

[0078] (5) Microscopic observation and photographing: same as in Example 1.

[0079] Experimental results: In this method, the soaking time of the sample is longer, the water content between the cells is larger, the tissue structure is looser, and the texture of the sample is too soft, which not only affects the control of the subsequent freezing time, but also may produce ice crystals in the cells. To some extent, the cell structure will be destroyed.

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Abstract

The invention provides a preparation method of frozen slices of traditional Chinese medicine cinnamomum cassia. The preparation method comprises the following steps of: material pretreatment: soaking a traditional Chinese medicine cinnamomum cassia sample in a solution containing 70% ethanol and water in a volume ratio of 1-2:1 for 1-2h; then taking out the traditional Chinese medicine cinnamomum cassia sample, and cutting the sample into short sections with a length of 5-8mm and a width of 5-8mm; soaking the short-section samples for 2-3h, taking out the short-section samples, wiping the short-section samples, putting the short-section samples into a test tube filled with a chloral hydrate test solution, and performing heating until the short-section samples are boiled for 5-7 times; then taking out the samples and wiping the samples; sample embedding, freezing and fixing; freezing and slicing; and flattening and slice making. According to the method, the tissue structure is not easy to deform, and the sample is not easy to break during slicing; dyeing is not needed; the traditional Chinese medicine cinnamomum cassia frozen slice with a complete structure can be quickly obtained; the slice tissue structure is clear, and cell inclusions are completely reserved; the manufacturing period is short, and the whole process from pretreatment to microscopic observation photographing can be completed within 4-6h, so that a complete image is obtained; the method is of great significance in identifying the traditional Chinese medicine cinnamomum cassia, expanding the application range of cinnamomum cassia and formulating related standards.

Description

technical field [0001] The invention relates to the technical field of microscopic tissue sectioning, in particular to a method for preparing frozen sections of traditional Chinese medicine cinnamon. Background technique [0002] Chinese medicine cinnamon is the dry bark of Cinnamomum cassia Presl, a plant of Lauraceae. It is very hot in nature, has the effects of nourishing fire and supporting yang, igniting fire and returning to the original, dispelling cold and relieving pain, and warming the meridians [1] . Cinnamon is a dual-purpose product for medicine and food. Its related products involve many fields such as medicine, food, daily necessities, and pesticides. It has a wide range of uses and a large amount of use. It is an important medicinal plant species in the world [2] . [0003] Since cinnamon and the counterfeit Chaigui are both Lauraceae plants, they have similar traits and smell, which can easily cause confusion [3-4] . And Chaigui is mainly used to treat ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/42G01N21/84
CPCG01N1/28G01N1/2813G01N1/42G01N21/84
Inventor 余佳琦孟千万沙禕炜吴清云
Owner 鉴甄检测技术(上海)有限公司
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