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Paraffin wax slicing method of ocean shellfish oocyte

A paraffin section and egg cell technology, which is applied to measurement devices, material analysis by optical means, instruments, etc., can solve problems such as unseen and small eggs, and achieve the effects of easy popularization and application, clear egg sections, and simple operation.

Active Publication Date: 2010-02-17
浙江万里学院宁海海洋生物种业研究院
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the traditional paraffin section technique has not solved many difficulties caused by small eggs, so there are few reports on the use of paraffin section technique to study the fertilization process of bivalve molluscs. haven't seen yet

Method used

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  • Paraffin wax slicing method of ocean shellfish oocyte
  • Paraffin wax slicing method of ocean shellfish oocyte

Examples

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Embodiment 1

[0036] 1. Sample fixation: Take meretrix meretrix from Shandong population, and conduct research on oxytocin and embryonic development at the Qingjiang Research Base of the Provincial Marine Aquaculture Research Institute in Wenzhou, Zhejiang. Take clam eggs of different development stages in 5mL centrifuge tubes, take samples every 5 minutes for 0-60 minutes after fertilization, and take samples every 10 minutes for 60-120 minutes, slowly add 2-3 mL of cold Bouin's fixative solution, and fix for 30 minutes. After the sample settles, replace the secondary fixative; fix with Bouin's solution for 5 hours, transfer to 70% alcohol, and store in a refrigerator at 4°C.

[0037] 2. Egg cell pretreatment: add 2 drops of 1% eosin dye solution to the centrifuge tube containing the egg cells, and stain for 1-2 minutes to make the surface of the egg cells dyed pink; take a square weighing paper and use a fine needle Puncture some small holes in the paper, then fold the paper, and wrap the...

Embodiment 2

[0044] The wide-shelled whole sea bamboo shoots are taken from the sea area of ​​Dongtou County, Wenzhou. After fertilization, fertilized eggs of different development stages are fixed. Since the diameter of the wide-shelled whole sea bamboo shoot eggs is only 40-50 μm, it is easy to lose when changing the fixative, so the amount of fixed eggs should be larger, and the centrifuge should be centrifuged at low speed to speed up the sedimentation.

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Abstract

The invention relates to a paraffin wax slicing method of ocean shellfish oocytes, comprising the steps of fixing, dehydration, clarity, waxdip, imbedment, slicing, dyeing, mounting, and the like. Theparaffin wax slicing method is characterized by solving the problem of the difficult waxdip and imbedment of the ocean shellfish oocytes due to small volume, improving a hematoxylin-eosin dyeing method and obtaining favorable slicing and dyeing effect. The paraffin wax slicing method is simple and easy to operate without adding instruments or equipment, can carry out paraffin wax slicing on mostof the ocean shellfish oocytes with the diameters more than or equal to 40 micrometers, transition steps are carried out among the dehydration, the clarity and the waxdip of the steps and mostly in two-step processing so that a reagent gradually and completely enters a tissue and is preserved in the original structure of the tissue in an intact way so as to provide an effective researching means for shellfish fertilization and early embryonic development research.

Description

technical field [0001] The invention relates to a paraffin sectioning method, in particular to a paraffin sectioning method suitable for marine shellfish egg cells with small egg diameters. Background technique [0002] Paraffin section technology is one of the important research methods of fertilization cytology. It has mature technology, stable method, low cost, convenient observation, and has the advantages of well-preserved tissue structure and clear image contrast, which can be clearly seen under light microscope. The changes in the external morphology and internal structure of fertilized eggs, especially the elongation and elongation of the spindle body and the changes in the nuclear membrane when the pronuclei of the two sexes combine, have unique advantages that are different from compression, transmission electron microscopy and fluorescence microscopy. Paraffin section technique has been widely used in fish fertilization cytology research. However, due to the smal...

Claims

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Application Information

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IPC IPC(8): G01N1/28G01N1/30G01N21/84
Inventor 董迎辉林志华
Owner 浙江万里学院宁海海洋生物种业研究院
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