Preparation method and application of Chinese giant salamander phalanx section
A technique of phalangeal and slicing, which is applied in the field of preparation of slices of phalanges of Chinese giant salamander, and can solve problems such as differences in results, incomplete generalization of methods for amphibians without tails and amphibians with tails, gaps in age identification, etc.
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Embodiment 1
[0027] A preparation method for Chinese giant salamander phalanx slices, the steps are as follows:
[0028] 1 Materials and methods
[0029] 1.1 Apparatus and reagents
[0030] Surgical scissors, tweezers, beaker, glass slides, cryostat (Leica, Germany), cryosection embedding agent OTC (Leica, Germany), staining cup.
[0031] 3% nitric acid solution, hematoxylin dye solution, eosin dye solution, 0.8% hydrochloric acid ethanol solution.
[0032] 1.2 Method
[0033] 1.2.1 Take the second joint of the giant salamander phalanx as the research object to make slices, and observe the age results of the giant salamander.
[0034] 1.2.2 Take giant salamander samples of known age and use this method for age identification.
[0035] Specific steps are as follows:
[0036] (1) Sampling: The toe bone samples of 4-year-old Chinese giant salamander were fixed in Bouin's solution (Bouin's) at room temperature for 24 hours. The formula of Born's solution is 75ml of picric acid, 25ml of a...
Embodiment 2
[0044] A preparation method of Chinese giant salamander phalanx slices, the steps are as follows:
[0045] 1 Materials and methods
[0046] 1.1 Apparatus and reagents
[0047] Surgical scissors, tweezers, beaker, glass slides, cryostat (Leica, Germany), cryosection embedding agent OTC (Leica, Germany), staining cup.
[0048] 3% nitric acid solution, hematoxylin dye solution, eosin dye solution, 0.8% hydrochloric acid ethanol solution.
[0049] 1.2 Method
[0050] 1.2.1 Take giant salamander phalanges as the research object to make slices and observe the age results of giant salamander.
[0051] 1.2.2 Take giant salamander samples of unknown age and use this method for age identification.
[0052] Specific steps are as follows:
[0053] (1) Sampling: The giant salamander phalanges of unknown age were fixed in Bouin's solution (Bouin's) at room temperature for 72 hours.
[0054] The formula of Born's solution is: mix 75ml of picric acid, 25ml of anhydrous formaldehyde and 3m...
Embodiment 3
[0061] A kind of application of the preparation method of Chinese giant salamander phalanx slice, its steps are as follows:
[0062] The slices prepared in Examples 1 and 2 were observed under a microscope, and the age was identified according to the number of annual ring marks on the slices.
[0063] From figure 1 It can be seen from the figure that, using the method of the present invention, it can be clearly seen that there is a group of concentric circles in the giant salamander skeleton. The concentric circles are blue-black after being stained with hematoxylin, and the outline is obvious but there will be incomplete closure. The circles The interphase is a deep blue-pink color displayed by both eosin and hematoxylin. The concentric circles are clearly identifiable, which is the so-called "bone age". The bone age of the known age identified by the method of the present invention is 4 years old; the bone age of the unknown giant salamander is 10 years old.
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