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
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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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