Method for fixing fish brain tissue specimen through systemic heart perfusion
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A technique for specimen fixation and brain tissue, applied in the biological field, can solve the problems of cardiac perfusion techniques that are rarely reported in literature, and achieve the effect of avoiding excessive bleeding and preventing shedding
Inactive Publication Date: 2015-04-29
YANSHAN UNIV
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[0012] Aquatic animals and terrestrial animals have great morphological and physiological
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Embodiment 1
[0032] Experimental animals: healthy adult carp, weighing 1.13kg and body length 36.52cm, purchased from the aquatic product market in Qinhuangdao City.
[0033] Main reagents: eugenol (Shanghai Medical Instrument Co., Ltd. Dental Material Factory, batch number 201103), normal saline, distilled water, neutral red, neutral gum, paraffin, xylene, absolute ethanol, 10% formalin solution , 2% hydrochloric acid solution.
[0034] Main instruments: BQ50S micro-flow speed-regulating peristaltic pump (Baoding Ref Fluid Technology Co., Ltd.), 7002 mold electric grinder (Yongkang Xiclair Tools Co., Ltd.), RM2235 tissue slicer (Germany Leica Company), TS100 inversion Biological microscope (Nikon Company, Japan), HH-S digital display constant temperature water bath (Jintan Medical Instrument Factory), 2 sets of infusion tubes, and 1 set of surgical instruments.
[0035] 1. Animal anesthesia: Mix 6 mL of eugenol and 48 mL of absolute ethanol and shake well, let it stand for 10 minutes, po...
Embodiment 2
[0045] Experimental animals: healthy adult crucian carp, weighing 0.33 kg and body length 19.21 cm, purchased from the aquatic product market in Qinhuangdao City.
[0046] Main reagents: eugenol (Shanghai Medical Instrument Co., Ltd. Dental Material Factory, batch number 201103), normal saline, distilled water, absolute ethanol, 10% formalin solution.
[0047] Main instruments: BQ50S micro-flow speed-regulating peristaltic pump (Baoding Ref Fluid Technology Co., Ltd.), 7002 mold electric grinder (Yongkang Xikelai Tools Co., Ltd.), 2 sets of infusion tubes, and 1 set of surgical instruments.
[0048] 1. Animal anesthesia: use 0.36g / L eugenol anesthesia solution for anesthesia, and the anesthesia time is about 5 minutes (the body is smaller, and the anesthesia time is shorter than that of carp), and it enters the state of anesthesia A3 (anesthesia phase III).
[0049] 2. Thoracotomy: The operation is the same as that in Embodiment 1, but because the crucian carp is small in size...
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Abstract
The invention discloses a method for fixing a fish brain tissue specimen through systemic heart perfusion. The method is mainly characterized in that after fish anesthesia, thoracotomy is performed on the periphery of a heart surface projection area of the fish, ventral aorta intubation is performed through the ventricle, common cardinal vein intubation is performed through the atrium, ligation with an operation thread is used for fixing, and an out flow tube is connected with a graduated vessel for measuring liquid out-flowing quantity; a constant-speed perfusion pump is used for perfusing normal saline through a perfusion tube, and a 10% formalin solution is adopted for reperfusion; when perfusion is started, a line is marked on the skull instantly and craniotomy is performed, and the perfusion time is then decided through combination of the perfusion quantity and the out-flowing quantity when the color of the brain tissue changes white; the brain is picked and placed in the 10% formalin solution to be fixed for 48 h after perfusion is stopped, and the brain tissue specimen with complete structure, normal form and good state is obtained. By means of the method, the problems of insufficient fixing, non-comprehensive fixing area, tissue autolysis, cell deformation and the like of the fish brain tissue can be solved effectively, the brain tissue specimen with more complete structure and more ideal state is obtained easily, and the natural form is retained better.
Description
technical field [0001] The invention belongs to the biological field, in particular to a method for fixing animal tissue samples. Background technique [0002] Cardiac perfusion is the technical basis for tissue sectioning, pathological specimen preparation, immunohistochemistry, in situ hybridization, in situ PCR, and intracerebral localization marker research on animal brain tissue. Animal in vivo heart perfusion technique is a method of fixation before tissue specimens, and it is a method of perfusing the heart with fixative solution when the animal is anesthetized. [0003] After death, animal brain tissue will soon undergo autolysis due to the action of intracellular lysosomal enzymes. The purpose of cardiac perfusion is to fix the brain tissue with fixative solution before animal death, inhibit the reproduction of bacteria in the tissue, and prevent corruption. , to avoid the occurrence of brain tissue autolysis, it can also increase the hardness of the tissue, make t...
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