Dehydrating and embedding improvement method of human body or animal tissues

A technology of animal tissue and dehydration bag, which is applied in the preparation of test samples, etc., can solve the problem of fragility of paraffin tissue blocks, and achieve the effect of less toxic and side effects, smooth tissue and high safety

Inactive Publication Date: 2013-08-07

THE THIRD XIANGYA HOSPITAL OF CENT SOUTH UNIV

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AI-Extracted Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the deficiencies of the prior art and provide an improved method of dehydration ...

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Abstract

The invention discloses a dehydrating and embedding improvement method of human body or animal tissues. The method comprises the following steps of: fixing; dehydrating; transparence treatment; wax-dipping and embedding. The method comprises the following steps of: first, dehydrating fixed tissue by using ethanol with different concentrations from low concentration to high concentration in a gradient manner; then, placing the tissue dehydrated in the gradient manner into a mixed solution of absolute ethyl alcohol and butanol in a volume ratio of 1:1-1:5 for dehydrating, wherein the time for dehydrating by using the mixed liquid is 30-60 minutes, and then, further using butanol to dehydrate the tissue which is dehydrated by using the mixed liquid at 4-16 DEG C for 8-20 hours; at the end of dehydration, dipping the tissue in butanol for transparence treatment; and finally, wax-dipping and embedding the transparent tissue. The method provided by the invention is simple and convenient to operate, high in safety and reliability, remarkable in effect and wide in application.

Application Domain

Preparing sample for investigation

Technology Topic

ButanolAlcohol ethyl +5

Examples

Experimental program(6)

Example Embodiment

[0018] Example 1:

[0019] In a method for improving dehydration and embedding of human or animal tissues of the present invention, skin tissue (can also be loose tissues such as glands and mucous membranes) is selected as the implementation object, and the specific operation steps are as follows:

[0020] 1. Fixation: Obtain skin tissue samples through clinical surgery or animal experiments, and quickly cut them into blocks of about 0.5cm×1cm×1cm (thickness×length×width), first use 4% paraformaldehyde (mass/volume ratio, solvent For PBS) fixation, overnight at 4℃, time is 8h-16h, and then placed in 0.02% (mass/volume ratio) NaN 3 Stored in the PBS storage solution for later use, can be stored for a long time.

[0021] 2. Dehydration: First, the fixed skin tissue sample is dehydrated by ethanol concentration gradient: the skin tissue sample is dehydrated by 50% volume concentration of ethanol for 20 minutes, 70% volume concentration of ethanol for 1 hour, and 80% volume concentration of ethanol for 1 hour. , 95% volume concentration ethanol dehydration for 45min, again with 95% volume concentration ethanol dehydration 45min, dehydrated ethanol dehydration 45min, complete the gradient dehydration, the gradient dehydration process is carried out at 4 ℃ ~ 16 ℃; after the gradient dehydration, the skin The tissue sample is placed in a 1:1 mixed solution of absolute ethanol and isobutanol for dehydration of the mixture. The dehydration time of the mixture is 45 minutes; after the mixture is dehydrated, put the obtained skin tissue sample in isobutanol Place it in a refrigerator at 4°C overnight for further dehydration, and the time is 14h-18h.

[0022] 3. Transparent: Put the dehydrated skin tissue sample into n-butanol and soak for 2h.

[0023] 4. Wax immersion: first immerse the skin tissue sample after transparent treatment in soft wax with a melting point of 45℃~56℃ for 1h, and then immerse it in hard wax with a melting point of 56℃~60℃ for 1h, set the first immersion The wax temperature is 55°C, and the second dip wax temperature is 60°C.

[0024] 5. Embedding: Put the waxed skin tissue sample at the bottom of the embedding frame, then add hard wax to the embedding frame and place it at 4°C overnight before sectioning.

Example Embodiment

[0025] Example 2:

[0026] An improved method for dehydration and embedding of human or animal tissues of the present invention selects liver tissue (also can be substantive normal tissues such as spleen, kidney, muscle, etc., or cancer tissue) as the implementation object. The specific operation steps are as follows :

[0027] 1. Fixation: Obtain liver tissue samples through clinical surgery or animal experiments, and quickly cut them into blocks of about 0.5cm×1cm×1cm, fix them with 4% paraformaldehyde, overnight at 4℃, for 8h-16h, then Put in 0.02%NaN 3 Stored in the PBS storage solution for later use, can be stored for a long time.

[0028] 2. Dehydration: First, the fixed liver tissue samples are dehydrated by ethanol concentration gradient: the liver tissue samples are dehydrated by 50% volume concentration of ethanol for 30 minutes, 70% volume concentration of ethanol for 1 hour to 2 hours, and 80% volume concentration of ethanol. Dehydrate for 1h~2h, dehydrated with 95% volume concentration of ethanol for 1h, again dehydrated with 95% volume concentration of ethanol for 1h, dehydrated with absolute ethanol for 1h, and dehydrated with absolute ethanol for 1h to complete the gradient dehydration. The gradient dehydration process is all at 4℃ ~16℃; after gradient dehydration, place the liver tissue sample in a 1:1 mixed solution of absolute ethanol and isobutanol for dehydration of the mixture. The dehydration time of the mixture is 1h; the mixture is dehydrated Afterwards, the obtained liver tissue samples were placed in isobutanol and placed in a refrigerator at 4°C overnight for further dehydration, for a period of 14 to 18 hours.

[0029] 3. Transparent: Put the dehydrated liver tissue sample into n-butanol and soak for 2h~4h.

[0030] 4. Wax immersion: first immerse the liver tissue sample after transparent treatment in soft wax with a melting point of 45℃~56℃ for 1.5h, then immerse it in hard wax with a melting point of 56℃~60℃ for 1.5h, set the first The second dip wax temperature is 55℃, and the second dip wax temperature is 60℃.

[0031] 5. Embedding: Put the liver tissue sample soaked in wax on the bottom of the embedding frame, then add hard wax to the embedding frame and place it at 4°C overnight before sectioning.

Example Embodiment

[0032] Example 3:

[0033] In a method for improving dehydration and embedding of human or animal tissues of the present invention, bone tissue is selected as the implementation object, and the specific operation steps are as follows:

[0034] 1. Fixation: Obtain bone tissue samples through clinical surgery or animal experiments, quickly cut them into blocks of about 0.5cm×1cm×1cm, fix them with 4% paraformaldehyde, overnight at 4℃, for 8h-16h, then Put in 0.02%NaN 3 Stored in the PBS storage solution for later use, can be stored for a long time.

[0035] 2. Dehydration: First, the fixed bone tissue samples are dehydrated by ethanol concentration gradient: the bone tissue samples are dehydrated by 50% volume concentration ethanol for 30 minutes, 70% volume concentration ethanol for 2 hours, and 80% volume concentration ethanol for 1 hour. , 95% volume concentration ethanol dehydration for 1 hour, again with 95% volume concentration ethanol dehydration for 1 hour, dehydrated ethanol dehydration for 1 hour, to complete the gradient dehydration, the gradient dehydration process is carried out at 4 ℃ ~ 16 ℃; after the gradient dehydration, the bone The tissue sample is placed in a 1:1 mixed solution of absolute ethanol and isobutanol for dehydration of the mixture. The dehydration time of the mixture is 1h; after the mixture is dehydrated, put the bone tissue sample in isobutanol Place it in a refrigerator at 4°C overnight for further dehydration, and the time is 14h-18h.

[0036] 3. Transparent: Put the bone tissue sample after dehydration into n-butanol and soak for 3h (3h~4h can be used).

[0037] 4. Wax immersion: first immerse the bone tissue sample after transparent treatment in soft wax with a melting point of 45℃~56℃ for 2h, and then immerse it in hard wax with a melting point of 56℃~60℃ for 1h, set the first immersion The wax temperature is 55°C, and the second dip wax temperature is 60°C.

[0038] 5. Embedding: Put the skeletal tissue sample soaked in wax on the bottom of the center of the embedding frame, then add hard wax to the embedding frame and place it at 4°C overnight before sectioning.

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Description & Claims & Application Information

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Classification and recommendation of technical efficacy words

Moderation
Improve safety and reliability

Dehydrating and embedding improvement method of human body or animal tissues

A technology of animal tissue and dehydration bag, which is applied in the preparation of test samples, etc., can solve the problem of fragility of paraffin tissue blocks, and achieve the effect of less toxic and side effects, smooth tissue and high safety

AI-Extracted Technical Summary

Problems solved by technology

Abstract

Examples

Example Embodiment

Example Embodiment

Example Embodiment

PUM

Description & Claims & Application Information

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