Method for collecting blood of small-size experimental fish
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A technology for blood collection and experimentation, which is applied to blood sampling devices, anticoagulation treatment, and types of packaging items. and destruction, preventing hemolysis, preventing platelet aggregation and thrombosis
Active Publication Date: 2020-05-05
GUANGDONG LAB ANIMALS MONITORING INST
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The above-mentioned operating device is complicated, which is not conducive to the rapid development of related experiments under laboratory conditions
In addition, due to the strict examination system for the welfare of experimental animals that limits the number of experimental animals used, the detection and evaluation of pathological models such as fat infiltration of small experimental fish often require dissection and tissue sampling during the modeling process. It is of practical significance to carry out testing and evaluation
At present, only the mullet goby (Mugilogobius chulae) and black-spotted medaka (Oryzias melastigma) are the only small seawater experimental fish developed internationally. In view of the huge differences in their living environments, species and morphological physiology, the research results of zebrafish Often not directly applicable to marine fish
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Embodiment 1
[0059] The mensuration of embodiment 1 blood collection and blood sugar content
[0060] A method for selecting a fish blood sampling location, comprising the following steps:
[0061] 1) Collect juvenile gobies fertilized for 96h–7d, stay in fresh water for 0.5h, wash with PBS 3-4 times, and fix in 4% paraformaldehyde overnight;
[0062] 2) After washing with PBST solution for 3 times, dehydrate with 25%, 50%, 75% and 100% methanol diluted with PBST in sequence, and incubate at room temperature for 15 min at each gradient concentration;
[0063] 3) Hydrate with 100%, 75%, 50% and 25% methanol diluted with NTMT solution in turn, and incubate at room temperature for 15 min at each gradient concentration; NTMT consists of 100 mM NaCl, 100 mM Tris-HCl, 50 mM MgCl2, 0.1% Tween 20 composition;
[0064] 4) After mixing NBT and BCIP (the mass ratio of NBT and BCIP: 100-450 μg / mL: 50-200 μg / mL), incubate at room temperature in the dark for 30 minutes, and then stop the reaction with...
Embodiment 2
[0091] Example 2, indicators such as glucose tolerance were used to compare the normal group of the mullet goby and the pancreatic fat infiltration model group.
[0092] The pancreatic fatty infiltration model goby in Example 2 of the present invention is the mullet goby Zhushi, which is a pancreatic fatty infiltration model fish prepared by a high-fat feeding method. It is compared with the pancreas slices of normal gobies Figure 8 . Figure 8 A is the pancreatic duct of CK normal goby; B is the pancreatic duct of the pancreatic fat infiltration model after long-term feeding with high-fat I high-fat diet.
[0093] See Table 2 for the determination of CK (control group) and high-fat feed ingredients.
[0094] Table 2 Feed basic composition analysis and particle size
[0095]
[0096] The model construction and specific feeding process are as follows: Strictly control the feeding conditions (12h light and night cycle, water temperature 24±2°C, pH 7.0±0.5, ammonia nitrogen...
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Abstract
The invention discloses a method for collecting blood of a small-size experimental fish. According to the method, an optimal blood collection position of mugilogobius chulae is confirmed through experiments such as early-stage blood vessel coloring, a capillary pipet which is applicable to rapid blood collection of the rimental fish within an appropriate range is prepared, and thus hemolysis and bacterial pollution can be effectively prevented. Meanwhile, the invention provides a solution formula which is particularly applicable to blood collection of a small-size seawater fish. By adopting the solution formula, platelet aggregation and thrombosis can be effectively prevented, an anticoagulation function can be achieved, blood glucose degradation can be well inhibited, and accuracy of sugar tolerance experiments can be ensured. On premise that survival of a model fish is ensured, the positive rate of a pancreas fat infiltration model can be evaluated through a method with the combination of sugar tolerance experiments and growth and blood indexes, and practical significances can be achieved.
Description
technical field [0001] The invention discloses a method for collecting blood of small seawater experimental fish. The invention relates to the fields of animal experiments and biotechnology, and relates to an improved blood collection device suitable for small experimental fish. Background technique [0002] In recent years, small experimental fish such as zebrafish (Danio rerio) and medaka (Oryzias latipe) have played an increasingly important role in genetics, environmental toxicology, pharmacology, molecular developmental biology, and immunology research. Mugilogobius chulae is currently the only standardized seawater experimental fish with independent intellectual property rights in China, and is recommended for biological toxicity experiments. Researchers have long been committed to the standardization of Gobies mullet. So far, two strains have been inbred to the 13th generation, and the closed population has been bred to the 19th generation. Systematic research has be...
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