目的 探索和研究安全、快速，且能长期保存人体蠕行螨标本的制作方法，以期满足人体寄生虫实验教学的需要。方法 采用透明胶纸粘贴法和挤刮涂片法获取蠕形螨，使用多种方法对人体蠕形螨进行处理，对比中性树胶、无味型封片剂以及甘油明胶的封片效果，在数码显微镜KoPa WiFi EDU软件系统下观察和拍摄。结果 背景杂质少油脂少不影响观察的标本可直接采用无味型封片剂或者甘油明胶对透明胶封片保存。透明胶未粘贴好导致气泡多或者油脂、汗毛多影响观察的标本可采用2%洗洁精短时间清洁后，依次放入浓度为75%～100%的乙醇溶液中脱水后，采用无味型封片剂封片保存。对于透明胶上密度较大的标本，可采用2%洗洁精长时间浸泡后，离心沉淀，取沉渣放入75%乙醇溶液中固定后，采用甘油明胶封片保存。结论 可采用无味型封片剂或者甘油明胶代替中性树胶作为蠕形螨标本制作的封片剂。采用2%洗洁精可清洗标本中存在的油脂且可达到从透明胶带中游离蠕形螨的效果。
Objective To establish a safe and rapid preparation method for long-term preservation of human Demodex specimen, and to meet the needs of experimental teaching of human parasitology. Methods Human Demodex mites were obtained by the cellophane tape method and the squeezing method and were treated with various methods to compare the sealing effect of neutral balsam, odorless mounting medium, and glycerol jelly mounting medium, and the morphology of mite was observed and photographed under the KoPa WiFi EDU software system of the digital microscope. Results The specimens with few background impurities or little grease which did not affect observation were directly sealed with odorless mounting medium or glycerol jelly mounting medium. In contrast, the specimens with excessive bubbles, grease, or hair which affected observation were first cleaned with 2% dish soap for a short period of time, then placed in 75%-100% ethanol for dehydration, and finally sealed with odorless mounting medium. If there was a high density of mites on one sheet of adhesive tape, such specimens were first soaked in 2% dish soap for a long time and then centrifuged, and the mites collected were fixed with 75% ethanol and sealed with a glycerol jelly mounting medium. Conclusion Odorless mounting medium or glycerol jelly mounting medium can replace neutral balsam as the mounting medium for preparing Demodex specimen, and 2% dish soap can remove the excess grease in specimen and easily release the mites from the adhesive tape.
[1] 卢致民,李凤铭. 临床寄生虫学检验技术[M]. 武汉:华中科技大学出版社,2019:213-214. Lu ZM,Li FM. Clinical parasitology and laboratory technology[M]. Wuhan:Huazhong University of Science & Technology Press,2019:213-214. (in Chinese)
[2] 蒲兴旺,刘刚,鲁开化. 人类蠕形螨虫与皮肤健康的临床研究进展[J]. 中国美容整形外科杂志,2018,29(8):510-511. DOI:10.3969/j.issn.1673-7040.2018.08.019. Pu XW,Liu G,Lu KH. Progress in clinical research on human Demodex and skin health[J]. Chin J Aesth Plast Surg,2018,29(8):510-511. DOI:10.3969/j.issn.1673-7040.2018.08.019.(in Chinese)
[3] 诸欣平,苏川. 人体寄生虫学[M]. 9版. 北京:人民卫生出版社,2018:245. Zhu XP,Su C. Human parasitology[M]. 9th ed. Beijing:People's Medical Publishing House,2018:245. (in Chinese)
[4] Hellerich U,Metzelder M. Incidence of scalp involvement by Demodex folliculorum Simon ectoparasites in a pathologic-anatomic and forensic medicine autopsy sample[J]. Arch Kriminol,1994,194(3/4):111-118.
[5] Madeira NG,Sogayar MIL. Prevalencia de Demodex folliculorum e Demodex brevis em Uma amostra da população de Botucatu,São Paulo,Brasil[J]. Rev Soc Bras Med Trop,1993,26(4):221-224. DOI:10.1590/S0037-86821993000400004.
[6] 李朝品. 医学蜱螨学[M]. 北京:人民军医出版社,2006:299-322. Li CP. Medical acarology[M]. Beijing:People's Military Medical Publishing House,2006:299-322. (in Chinese)
[7] 郭家,郭俊杰,张浩,等. 制备蠕形螨玻片标本的新方法:透明胶纸永久封片法[J]. 齐齐哈尔医学院学报,1999,20(5):507-508. Guo J,Guo JJ,Zhang H,et al. A new method for preparing Demodex glass slides:permanent mounting cellophane tape[J]. J Qiqihar Med Univ,1999,20(5):507-508. (in Chinese)
[8] 王唯唯,黄文达. 中性树胶直接封片制作蠕形螨标本[J]. 中国寄生虫学与寄生虫病杂志,1998,16(2):76. Wang WW,Huang WD. Preparation of Demodex specimen by direct mounting with neutral gum[J]. Chin J Parasitol Parasit Dis,1998,16(2):76. (in Chinese)
[9] 李永祥,张辉,刘振忠,等. 蠕形螨标本制作方法探讨[J]. 中国寄生虫学与寄生虫病杂志,2002,20(4):196. DOI:10.3969/j.issn.1000-7423.2002.04.018. Li YX,Zhang H,Liu ZZ,et al. Preparation and staining of Demodex folliculorum specimen[J]. Chin J Parasitol Parasit Dis,2002,20(4):196. DOI:10.3969/j.issn.1000-7423.2002.04.018.(in Chinese)
[10] 李永祥,刘振忠,郑志红. 蠕形螨标本的制作与染色[J]. 中国人兽共患病杂志,2002,18(2):12. DOI:10.3969/j.issn.1002-2694.2002.02.048. Li YX,Liu ZZ,Zheng ZH. Demodicidae sample making[J]. Chin J Zoonoses,2002,18(2):12. DOI:10.3969/j.issn.1002-2694. 2002.02.048.(in Chinese)
[11] 刘莹,李速婷. 人体蠕形螨环境扫描电镜样本的制作[J]. 医学动物防制,2007,23(11):818-819. DOI:10.3969/j.issn.1003-6245.2007.11.008. Liu Y,Li ST. Preparation of environmental scanning electron microscope samples of human Demodex[J]. Med Ani Prev,2007,23(11):818-819. DOI:10.3969/j.issn.1003-6245.2007.11.008.(in Chinese)
