Fluorescent dyeing method for cuttlefish embryonic development and nuclear action observation

A technology of embryonic development and fluorescent dyeing, applied in the field of fluorescent dyeing, can solve the problems of destroying materials and structures, deforming eggs, affecting the objectivity of observation results, etc., and achieves the effects of simple steps, convenient observation, and convenient operation

Inactive Publication Date: 2009-07-29
ZHEJIANG OCEAN UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

When the researchers observed the embryos of squid larvae, because the color of squid ink was black during the microscopic examination, it would seriously affect the accuracy of the naked eye microscopic examination, so a new method was needed to solve this problem. The nak

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  • Fluorescent dyeing method for cuttlefish embryonic development and nuclear action observation

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Embodiment 1

[0025] Example 1: Fluorescence staining method for the observation of embryonic development and nuclear behavior of squid, which has the following steps:

[0026] ①Select squid eggs, peel off the outer egg membrane and only keep the innermost egg membrane;

[0027] ②The cuttlefish eggs treated in step ① are immersed in the fixative for storage at 4°C, and the fixative is replaced 3 times during storage;

[0028] ③ Wash the cuttlefish eggs fixed in step ② with buffer solution for 3 times, and then dye them with fluorescent dyes, and the dyeing process is carried out in a dark environment;

[0029] ④ Cut off the tiny animal heads of the squid eggs processed in step ③, spread them on a glass slide, and take pictures and image under a fluorescence microscope.

[0030] In step ②, the fixing solution contains 2 parts of glutaraldehyde, 2.5 parts of paraformaldehyde, and 95.5 parts of filtered seawater by weight.

[0031] The dye in step ③ is 4,6-diamidinophenylindole, and the dyeing tim...

Embodiment 2

[0035] Example 2: Fluorescence staining method for the observation of embryonic development and nuclear behavior of squid, which has the following steps:

[0036] ①Select squid eggs, peel off the outer layer of egg membrane and retain only the innermost layer of egg membrane; squid eggs are large in size, ellipsoid-shaped and covered with three layers of egg membranes, cell division and development occur at the animal pole (small egg head), The outer egg membrane has a compact structure, which is not conducive to the penetration of the fixing solution used in the method of the present invention. Therefore, the two outer egg membranes should be removed, and only the innermost egg membrane should be retained; the other of the innermost egg membrane should be retained. One advantage is that the egg will not deform, and will not damage the material and structure existing on the surface of the fertilized egg, ensuring the accuracy of the observation results.

[0037] ②The cuttlefish eg...

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Abstract

The invention discloses a fluorescent staining method of cuttlefish embryo development and nuclear behavior observation. The method comprises the following steps: (1) selecting a cuttlefish egg, peeling off outer egg membranes and reserving an innermost egg membrane; (2) submerging the cuttlefish egg treated in the step (1) in a fixing liquid for conservation at 4 DEG C, and changing the fixing liquid for twice to four times during the conservation period; (3) washing the cuttlefish egg fixed in the step (2) with buffer solution for twice or thrice, and staining with a fluorescent staining agent in dark environment; and (4) cutting off a capitulum at the animal pole of the cuttlefish egg treated in the step (3), laying on a slide glass, and imaging under a fluorescence microscope. The method has the advantages of not destroying substances and structures on the surface of fertilized eggs, is characterized by simple step and convenient operation, and can rapidly, clearly and directly observe embryonic development and nuclear behavior of the cuttlefish under the fluorescence microscope.

Description

Technical field [0001] The invention relates to a fluorescent dyeing method for observation of the development and nuclear behavior of aquatic animals, in particular to a fluorescent dyeing method for the observation of embryonic development and nuclear behavior of squid. Background technique [0002] Cuttlefish, commonly known as cuttlefish, belong to the order of the cephalopods and ten arms. Squid meat contains a variety of amino acids, rich in nutrients and delicious meat. Squid ink has blood coagulation effect and is mainly used to stop bleeding and treat blood diseases; it can also protect hematopoietic stem cells, contains anti-cancer substances and has anti-tumor activity; cuttlefish bones are called cuttlefish, which have astringent, antacid, and hemostatic effects. One of the four major fishery targets in my country, it is highly adaptable to temperature and is widely distributed along the coast of China. It is a traditional fishery species with high economic value. Due...

Claims

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Application Information

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IPC IPC(8): G01N21/64
Inventor 吴常文迟长凤徐佳晶
Owner ZHEJIANG OCEAN UNIV
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