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Tissue section cutting method for sea water left eye floundre and right eye founder fertilized egg

A technology of tissue slices and fertilized eggs, which is applied in the preparation of test samples, instruments, teaching models, etc., to achieve the effect of avoiding operation steps, high repeatability and good effect

Inactive Publication Date: 2006-03-22
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the inadequacy that the slicing method of fertilized eggs of common organisms and freshwater fish cannot be applied to the slicing of seawater flounder and flounder eggs, the present invention has established a whole set of Material Fixation, Handling and Tissue Sectioning Methods for Saltwater Flounder Roe Slicing

Method used

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  • Tissue section cutting method for sea water left eye floundre and right eye founder fertilized egg
  • Tissue section cutting method for sea water left eye floundre and right eye founder fertilized egg
  • Tissue section cutting method for sea water left eye floundre and right eye founder fertilized egg

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Example 1: Tissue section of flounder fertilized eggs

[0026] Step 1: put the fertilized eggs of flounder at different development stages into 50 times the volume of Bouin's fixative solution for 10 hours, wash with 70% ethanol for several times (wash away the fixative solution) and save.

[0027] Step 2: After the preserved eggs are washed with 70% ethanol, the egg membrane is punctured in the direction of the plant pole with a dissecting needle.

[0028] Step 3: Place it on a glass slide, and adjust the position and direction of the egg under a dissecting microscope after dripping hot-melt agar.

[0029] Step 4: Dehydrate the embedded agar block as a tissue block with 70%, 80%, 90%, 95%, and 100% ethanol, and the treatment time for each group is 30 minutes. A small amount of eosin was added to 95% ethanol to stain the ovules red.

[0030] Step 5: Put it into terpineol for transparent treatment, and the treatment time is 4 hours.

[0031] Step 6: Put terpineol and ...

Embodiment 2

[0035] Example 2: Tissue section of turbot fertilized eggs

[0036] Step 1: Put fertilized turbot eggs at different development stages in 50 times the volume of Bouin's fixative solution for 12 hours, wash with 70% ethanol for several times (wash away the fixative solution) and save.

[0037] Step 2: After the preserved eggs are washed with 70% ethanol, the egg membrane is punctured in the direction of the plant pole with a dissecting needle.

[0038] Step 3: Put it on a glass slide, drop hot-melt agar and adjust the position and direction of the eggs under a dissecting microscope.

[0039] Step 4: Dehydrate the embedded agar block as a tissue block with 70%, 80%, 90%, 95%, and 100% ethanol, and the treatment time for each group is 1 hour. A small amount of eosin was added to 95% ethanol to stain the ovules red.

[0040] Step 5: Put the dehydrated agar block into terpineol for transparent treatment, and the treatment time is 6 hours.

[0041] Step 6: Put terpineol and hot-m...

Embodiment 3

[0045] Example 3: Tissue section of flounder fertilized eggs

[0046] Step 1: Put the fertilized eggs of flounder at different development stages in 100 times the volume of Bouin's fixative solution and fix them for 6 hours, wash them with 70% ethanol for several times (wash away the fixative solution) and save them.

[0047] Step 2: After the preserved eggs are washed with 70% ethanol, the egg membrane is punctured in the direction of the plant pole with a dissecting needle.

[0048] Step 3: Put it on a glass slide, drop hot-melt agar and adjust the position and direction of the eggs under a dissecting microscope.

[0049] Step 4: Dehydrate the embedded agar block as a tissue block with 70%, 80%, 90%, 95%, and 100% ethanol, and the treatment time for each group is 45 minutes. A small amount of eosin was added to 95% ethanol to stain the ovules red.

[0050] Step 5: Put the dehydrated agar block into terpineol for transparent treatment, and the treatment time is 12 hours.

...

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Abstract

The present invention relates to microscopic tissue section technology, and establishes one effective tissue section method for the fertilized egg of sea water left eyed founder and right-eyed founder based on the features of the fertilized egg of sea water left eyed founder and right-eyed founder. The fertilized egg is polylecithal pelagic egg with high egg membrane toughness and narrow perivitelline space. The tissue section method includes the following steps: 1. fixing the polylecithal pelagic egg with Bouiní»s fixing fluid and optimizing treatment period; 2. pricking the egg membrane with dissecting needle in the vegetative pole direction to peel off the egg membrane; and 3. directly embedded to fix embryo with hot agar. The method is simple and effective, and may be also applied in the tissue section of other sea water fish egg.

Description

technical field [0001] The invention relates to a micro-tissue slicing technique, in particular to a set of embryo positioning embedding continuous slicing method suitable for fertilized eggs of marine flounder and flounder fish. technical background [0002] In the existing reports on fish roe sectioning, the materials used are freshwater fish roe, such as grass carp and crucian carp, while the tissue section of seawater fish roe has not been reported yet. For freshwater fish eggs, fix them with Bouin's fixative solution or Smith fixative solution, remove the film, dehydrate them with ethanol gradient according to the common biological tissue section method, and make paraffin-embedded sections after transparent treatment with xylene. [0003] But utilize this method to carry out the tissue slice of marine flounder flounder roe but be easy to chop, can not obtain complete slice. The main reason is that flounder flounder eggs are different from most other freshwater fish egg...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/06G09B23/28
Inventor 孙威尤锋张培军徐永立
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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