超声加速蔗糖对动物组织脱水过程的可行性探索

摘要

Objective To examine the feasibility of enhancement of dehydration of animal tissues using sucrose so -lution.Methods SD rats were transcardically perfused to collect spinal cord samples for fixation .These obtained sam-ples were divided into two groups:a control group subjected to routine dehydration procedures and an ultrasound treatment group which underwent ultrasound enhanced dehydration for 10 min or 30 min.Then, all samples were cut into frozen sections for H -E staining .Meanwhile , the effects of ultrasound enhanced dehydration on immunofluorescence results were observed.Results According to H-E staining, 10 to 30 min of ultrasound enhanced dehydration still caused ice crystal holes in frozen sections with 0.3-0.5 mm thickness.Based on the background of immunofluorescence , these holes might not appear if the samples were cut into 1/3 of their initial dimensions before ultrasound enhanced dehydration for 10 min.Moreover , after ultrasound enhanced dehydration , the samples could be stained with ionized calcium binding adaptor molecule 1 (Iba1) and glial fibrillary acidic protein (GFAP) by immunofluorescence, which were comparable with the control .Conclusion Ultrasound can enhance the dehydration of animal tissues in sucrose solution , which can be then applied for immunofluorescence .%目的：检测超声是否可以加速蔗糖对动物组织的脱水过程。方法 SD大鼠经心脏灌注固定并取材脊髓组织，组织经后固定后分为2组，对照组进行常规蔗糖脱水，超声加速脱水组进行超声加速蔗糖脱水10 min或者30 min。对2组标本均进行冰冻切片，经苏木精－伊红染色，观察组织空洞存在情况以反映组织脱水的程度，并做免疫荧光染色观察超声加速脱水是否会对荧光染色的结果造成影响。结果苏木精－伊红染色结果显示，对0．3～0．5 mm完整的脊髓组织10～30 min超声加速脱水处理均不能避免冰冻后空洞形成。从免疫荧光染色背景判断，剪裁至原来1／3大小后的脊髓组织经超声加速脱水10 min可避免脊髓冰冻后的空洞形成。免疫荧光染色显示超声加速脱水组织可以进行离子钙接头蛋白1（ Iba1）和神经胶质纤维酸性蛋白（ GFAP）免疫荧光染色，且染色效果和对照组之间没有明显区别。结论超声可加速蔗糖对动物组织的脱水过程，可用于组织的免疫荧光染色。