不同脱钙条件下三种脱钙液脱钙效果的比较

Comparison for decalcifying effect of three decalcification reagents in the bone tissue

目的探讨在常温和加温脱钙条件下,甲酸混合液、Alexander硝酸甲醛液及Jenking脱钙液的脱钙效果和染色结果。方法取小鼠膝关节骨组织分为6份,根据脱钙条件和脱钙液的不同,分为常温脱钙组合加温脱钙组两组。常温脱钙组:三种脱钙液常温(25℃)脱钙48h并石蜡切片HE染色;加温脱钙组:三种脱钙液70℃温箱脱钙6h并石蜡切片HE染色,并分别观察骨组织在不同脱钙液及脱钙条件下的脱钙效果及染色结果。结果常温(25℃)脱钙组标本中,经甲酸混合液脱钙的骨组织较难切片,染色较差;经Alexander硝酸甲醛液脱钙的骨组织较易切片,染色较差;经Jenking脱钙液脱钙的骨组织较易切片,染色较好。加温脱钙组标本中,经甲酸混合液脱钙的骨组织较易切片,染色较好;经Alexander硝酸甲醛液脱钙的骨组织较易切片,染色一般;经Jenking脱钙液脱钙的骨组织较易切片,染色较好。结论不同的脱钙液种类及脱钙条件对脱钙及染色结果有较大的影响,标本经三种脱钙液加温脱钙后,使脱钙时间大大缩短,减少了酸对组织的破坏,在脱钙效果及染色结果方面,均要优于常温脱钙,而Jenking脱钙液无论在常温和加温条件下的脱钙和染色效果均要优于其它两种脱钙液,有一定的应用价值。

Objective To compare decalcification and subsequent staining effects of 20% formic acid, Alexander nitric acid-formaldehyde solution and Jenking decalcification solution under normal and increased temperature. Method Mouse knee joint bone tissues were obtained and divided into 6 groups： 3 decalcification agents under either room temperature （25°C） or increased temperature （70°C）. The decalcification lasted 48 and 6 hours for normal and increased temperature groups respectively, followed by paraffin section and HE staining. The decalcification efficiency and staining results were evaluated. Result Among those that were decalcified under normal temperature, bone tissues were most difficult to slice in 20% formic acid treated group and HE staining worked best in Jenking decalcification solution treated group. With increased temperature, bone tissues were easier to slice in all groups than they were under normal temperature. Overall the HE staining was good except in Alexander nitric acid-formaldehyde solution group. Conclusion Different decalcification agents and conditions directly influence decalcification efficiency and staining effects. Increasing temperature not only shortened decalcification time and reduced tissue damage by acids, but also showed better decalcification and staining results than normal temperature. Jenking decalcification solution, which was superior in all criteria, had potential application value.