【摘要】 在介绍制作新鲜冰冻神经纤维切片简单、有效方法的同时,报告了在切片上同时显示三磷酸腺苷酶(ATP-ase)及胆碱酯酶(ChE)活性的组织化学方法,这对观察神经纤维内两种酶的相互关系和生理或病理性的细胞化学改变是有意义的。更多还原

【Abstract】 The histochemical method of cutting nerve fiber lengthways andshowing the activities of ATPase and ChE simultaneous on the same tissuesection was studied.It can not only distinguish the localization of the twoenzymes,but also reflect the vitalities and functions of them.The procedure isas follows:1).Draw the fresh nerve-muscle specimen slightly to its naturallength,freeze it speedly,then cut it lengthways and successively（10μm）until thevertical sections could been seen.Put the sections into an exsiccator for 10 to 15min with negative pressure,then fixed them for 20 min with 1% paraformal-dedyle（pH=7.4）at room temperature and wash them 3 to 5 times withcold water.2).Put the sections into ATPase solution for 1-2 hours at 37℃,wash 3 times with distilled water,then steep them in 1%CaCl₂for 3 times,2% CoCl₂ for 3 min,1% （NH₄）₂ S for 0.5～1 min respectively,and washthem 3 times with distilled water.3).Put the sections into ChE solution for 2—4 hours at 37℃,wash them 3 times with distilled water,have them passthrough alcolhol of different concentration,make them transparent with xylene andseal them up with Canada Gum.ATPase Solution.0.1M Sodium barbiturate 10ml0.18M Calcium Chloride 5mlDistilled Water 15mlAdenosin-5’-TriphosphatDisodium Salt 76mgWhen ATP is dissolved completely,adjust the solutionto pH7.4 with 1N NaOH andadd distilled water to total volume 50ml,then filte the solution.ChE Solution:Solution A:0 Aceto-5-Bromindoxyl 1.3mgAlcolhol 0.1mlSolution B:0.1M Tras/HCl Buffer（pH6-8） 2ml0.05M Potassium Ferricyanide 1.0ml0.05M Potassium Ferrocyanide 1.0ml0.1M Calcium Chloride 1.0mlDistilled Water 5.0mlMix the solution A and B just before the test.更多还原