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作 者:何韶衡[1]
机构地区:[1]汕头大学医学院变态反应学与炎症学研究所,广东汕头515031
出 处:《汕头大学医学院学报》2002年第3期131-132,共2页Journal of Shantou University Medical College
摘 要:目的 :介绍哮喘患者支气管肺泡灌洗液 (BALF)的处理方法和肥大细胞计数。方法 :来源于稳定期轻度哮喘患者 (36人 )的BALF经过滤和冲洗后 ,取 10 μL与等量的 0 1%锥虫蓝混合并计数有核细胞总数。根据计数的结果将余下的细胞稀释成 5× 10 5有核细胞 mL ,然后进行细胞离心 ,每张载玻片上加 5× 10 4 个有核细胞。肥大细胞经Carnoy s固定溶液固定后 ,用甲苯胺蓝染色 ,并在脱水后用DPX封片。每张片子至少计数 5 0 0个有核细胞 ,并计数肥大细胞占有核细胞的百分率。结果 :肥大细胞呈深紫色 ,在淡蓝色背景上清晰易辨。稳定期轻度支气管哮喘患者的BALF中的肥大细胞占有核细胞的 (0 2 0± 0 2 4 ) %。结论 :本文的实验程序适合于哮喘患者BALF中的肥大细胞计数。Objective:To develop a procedure for processing bronchoalveolar lavage fluid(BALF)and counting mast cells of asthmatics.Methods:BALFs recovered from 36 mild asthmatics were filtered and washed.For counting total numbers of cells,10 μL of cell suspension was mixed with the equal volume of 0 1% trypan blue solution for 10 minutes before counting under microscope.Based on the numbers of cells counted,the cell suspension was diluted to 5×10 5 nucleated cells/mL.Cytospin slides were then prepared with 5×10 4 nucleated cells per slide.Following fixed in Carnoys fixative solution,mast cells were stained with 0 5% toluidine blue.A minimum of 500 cells per slide was counted and the percentage of mast cells calculated.Results:Mast cells appeared in dark purple,and the rest of cells stained in light blue.Mast cells consist of (0 20±0 24)% total nucleated cells in BALF.Conclusion:The experimental procedures demonstrated are appropriate for evaluation of numbers of mast cells in BALF.
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