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微波固定:蜱(蜱螨亚纲:硬蜱科)的原位组织解剖、蜱组织的薄切片以及小鼠脾脏中抗原的保存

Microwave fixation: in situ tick (Acari: Ixodidae) histoanatomy, thin sectioning of tick tissues, and antigen preservation in mouse spleen.

作者信息

Carranza G A, Cruz D, Wagner G G

机构信息

Department of Veterinary Microbiology and Parasitology, College of Veterinary Medicine, Texas A&M University, College Station 77843.

出版信息

J Med Entomol. 1990 Nov;27(6):1067-70. doi: 10.1093/jmedent/27.6.1067.

Abstract

Microwave irradiation was used for the fixation of eggs, nymphs, and adult Boophilus spp. ticks. Although optimal temperatures for fixation of the different tick stages varied, heating to 58 degrees C of adult ticks submerged in either PBS or fixative was found to be sufficient. After microwave fixation, whole adult ticks, hand held, were sectioned with a sharp razor blade. The resulting sections revealed the in situ histoanatomy of the tick. Thin sections of ticks were obtained after either paraffin or polyester wax embedding. Microwave fixation combined with polyester wax embedding made serial thin sections of the different stages of Boophilus ticks possible. The technique preserved antigens as demonstrated by the immunostaining of lymphocytes and erythrocytes infected with Babesia microti in mouse tissues subjected to the same treatment as the ticks. With the microwave fixation-polyester wax technique, the specimen preparation time from fixation to the section on the glass slide was reduced to less than 8 h.

摘要

微波辐照用于固定微小牛蜱属蜱虫的卵、若虫和成虫。尽管不同蜱虫阶段的最佳固定温度有所不同,但发现将浸没在磷酸盐缓冲盐水(PBS)或固定剂中的成年蜱虫加热到58摄氏度就足够了。微波固定后,手持完整的成年蜱虫,用锋利的剃须刀片切片。所得切片揭示了蜱虫的原位组织解剖结构。蜱虫的薄切片是在石蜡或聚酯蜡包埋后获得的。微波固定与聚酯蜡包埋相结合,使得制作微小牛蜱不同阶段的连续薄切片成为可能。如对与蜱虫接受相同处理的小鼠组织中感染微小巴贝斯虫的淋巴细胞和红细胞进行免疫染色所示,该技术保存了抗原。采用微波固定 - 聚酯蜡技术,从固定到在载玻片上切片的标本制备时间缩短至不到8小时。

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