摘要
目的:常规的脱钙骨切片制作方法在脱钙、组织结构完整性等方面存在一定的缺陷。比较用改良后方法制作的脱钙骨组织切片与常规制作的脱钙骨组织切片质量的差异。方法:①实验材料:实验于2004-12/2007-12在广东医学院组织和胚胎学教研室及药理教研室完成。SPF级雌性SD大鼠10只,鼠龄6个月。②实验方法:取大鼠左右股骨和胫骨标本各20个,分为两组,左股骨和右胫骨为实验组,右股骨和左胫骨为对照组。实验组经过常规固定,而后应用改良脱钙液(甲酸10mL、浓盐酸(相对密度1.19)8mL、氯化铝3g、体积分数为0.4的甲醛20mL、冰乙酸3mL,加生理盐水至总体积200mL)脱钙、脱水、氯仿透明、包埋时间延长、染色、封固和切片;对照组进行常规固定、脱钙、脱水、透明、包埋、染色、封固和切片。③实验评估:比较两组切片的质量和效果。结果:①实验组大鼠的脱钙骨切片与对照组比较,标本脱钙时间缩短,脱水时间变化不大,而透明效果明显改善。②与对照组比较,实验组骨组织切片标本薄均匀、平坦完整,脱蜡完整均匀。染色成功的切片,骨与骨周围的组织形态保持完整。结论:改良的脱钙大鼠股骨和胫骨石蜡切片制作方法能使标本脱钙更彻底、结构更清晰、组织完整性更好。
AIM： There is disadvantage in the conventional method of making bone decalcification and complete sections. This study compared the difference of making bone decalcification sections technique between the modified and conventional methods.
METHODS： ①Experiments were performed at the Department of Histology and Embryology, Department of Pharmacology, Guangdong Medical College from December 2004 to December 2007. Ten female SD rats of SPF grade aged 6 months were selected ②Totally 20 thigh bone and 20 tibia of rats were divided into 2 groups as experimental group （left thigh bone and fight tibia） and control group （right thigh bone and left tibia）. Rats in the experimental group received conventional fixation, and then decalcified with modified decalcifying fluid [（10 mL of methanoic acid, 8 mL of concentrated hydrochloric acid （1.19 relative density）, 3 g of aluminum chloride, 20 mL of formaldehyde （0.4 volume fraction）, 3 mL of glacial acetic acid, adding to 200 mL with saline solution）], dehydrated, transparentized with chloroform, and then embedded for a long time, dyed, mounted and made into tissue sections. ③Sections quality and effects were compared between both groups.
RESULTS： ①Decalcified sections were compared between both groups. Decalcification duration was shorter, but dehydration duration did not change. Transparent effects were improved. ②Compared to the control group, the section samples were even, thin, integrated, the deparaffinage was complete and even, and the bone and surrounding tissues were integrated in stained sections in the experimental group.
CONCLUSION： Decalcification is thorough, structure is clear and tissue integrity is better by the modified method of making rat thigh bone and tibia sections.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第11期2125-2128,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
湛江市科技计划资助项目(2007C02011)~~
