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A fixative solution for eyeball tissue and a pretreatment method for making slices of eyeball tissue

A fixative and pretreatment technology, which is applied to the pretreatment of eyeball tissue production and the field of eyeball tissue fixative, can solve the problems of the fixative solution being difficult to penetrate the dense sclera, increasing the difficulty of eyeball production, and preparing eyeball paraffin sections, etc. , to achieve the effect of clear tissue structure without cracks, maintaining structural integrity, preventing fragmentation and shedding

Active Publication Date: 2022-01-18
武汉赛维尔生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

During the fixation process, it is difficult for conventional fixatives to penetrate the dense sclera, making it impossible to use the existing paraffin section method to prepare eyeball paraffin sections
Paraformaldehyde is often used to fix first and then dehydrated with ethanol, but due to the shrinkage of each layer, tissue separation will occur, especially the separation and detachment of the retina
[0003] In addition, due to the presence of aqueous humor and vitreous body in the eyeball to maintain the pressure inside the eyeball, this further increases the difficulty of eyeball filmmaking. If the operator directly removes the aqueous humor and vitreous body through rough cutting, it is easy to cause uneven incisions and even damage Structure of the eyeball, causing retinal detachment and fragmentation

Method used

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  • A fixative solution for eyeball tissue and a pretreatment method for making slices of eyeball tissue
  • A fixative solution for eyeball tissue and a pretreatment method for making slices of eyeball tissue
  • A fixative solution for eyeball tissue and a pretreatment method for making slices of eyeball tissue

Examples

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Effect test

Embodiment 1

[0030] The components of the fixative solution for eyeball tissue are as follows: 35 mL isotonic aqueous humor, 10 mL formaldehyde, 5 mL acetic acid, 37.5 mL absolute ethanol, and 12.5 mL pure water. The isotonic aqueous humor is composed of sodium chloride, urea, potassium chloride and pure water, the concentration of sodium chloride is 137.9mM, the concentration of urea is 13.43mM, and the concentration of potassium chloride is 2.67mM. The above liquids were mixed to obtain a colorless and precipitate-free solution, which was the fixative solution of this example.

[0031] A pretreatment method for eyeball tissue preparation, comprising the steps of:

[0032] (1) using the fixative to fix the eyeball tissue for 22 hours;

[0033] (2) using OCT embedding agent to embed the fixed eyeball tissue in step (1);

[0034] (3) freezing the eyeball tissue embedded in step (2) at -20°C;

[0035] (4) Put the frozen eyeball tissue obtained in step (3) on a cryostat at -20°C, cut off 1...

Embodiment 2

[0051]The components of the fixative solution for eyeball tissue are as follows: aqueous humor isotonic solution 40mL, formaldehyde 7mL, acetic acid 3mL, absolute ethanol 30mL, pure water 20mL. The isotonic aqueous humor consists of sodium chloride, urea, potassium chloride and pure water, wherein the concentration of sodium chloride is 150 mM, the concentration of urea is 3 mM, and the concentration of potassium chloride is 1 mM. The above liquids were mixed to obtain a colorless and precipitate-free solution, which was the fixative solution of this example.

[0052] A pretreatment method for eyeball tissue preparation, comprising the steps of:

[0053] (1) using the fixative to fix the eyeball tissue for 24 hours;

[0054] (2) using OCT embedding agent to embed the fixed eyeball tissue in step (1);

[0055] (3) freezing the eyeball tissue embedded in step (2) at -25°C;

[0056] (4) Place the frozen eyeball tissue obtained in step (3) on a cryostat at -25°C, cut off 1 / 3 of...

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Abstract

The invention belongs to the technical field of pathological reagents, and in particular relates to a fixative solution for eyeball tissue and a pretreatment method for preparing eyeball tissue slices. The components of the fixative are calculated by volume percentage: 20-50% aqueous humor isotonic fluid, 1%-10% acetic acid, 1-15% formaldehyde, 30%-50% absolute ethanol, 10-50% pure water 20%; the isotonic liquid of described aqueous humor is made up of sodium chloride, urea, potassium chloride and pure water, and the concentration of sodium chloride is 136~154mM, and the concentration of urea is 10~15mM, and the concentration of potassium chloride is 1 ~3mM. Using the fixative and pretreatment method of the present invention to make slices can ensure the integrity of the morphological structure of the eyeball, can prevent the retina from breaking and falling off in the slice making process, and is useful for eyeball follow-up morphology, histology, and pathology research. Positive and profound meaning.

Description

technical field [0001] The invention belongs to the technical field of pathological reagents, and in particular relates to a fixative solution for eyeball tissue and a pretreatment method for preparing eyeball tissue slices. Background technique [0002] The structure of the eyeball is precise and complex, the softness and hardness of the various layers of the eyeball wall vary greatly, and the connectivity between the retina, choroid, and sclera is poor. During the fixation process, it is difficult for conventional fixatives to penetrate the dense sclera, making it impossible to use the existing paraffin section method to prepare ocular paraffin sections. Paraformaldehyde is often used to fix first and then dehydrated with ethanol, but due to the shrinkage of each layer, tissue separation will occur, especially the separation and detachment of the retina. [0003] In addition, due to the presence of aqueous humor and vitreous body in the eyeball to maintain the pressure in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/28G01N1/30G01N1/36G01N1/42G01N1/06
CPCG01N1/286G01N1/30G01N1/36G01N1/42G01N1/06G01N2001/305G01N2001/2873
Inventor 张高英刘姗姗洪盼
Owner 武汉赛维尔生物科技有限公司
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