汕头大学医学院学报2025，Vol.38Issue(1)：32-35,4.DOI:10.13401/j.cnki.jsumc.2025.01.007
免疫荧光技术中不同固定方式及渗透时间对人红细胞带3蛋白染色效果的影响
Effect of different fixation methods and penetration time on the staining effect of human erythrocyte band 3 protein in immunofluorescence technique
摘要
Abstract
Objective:To investigate the effects of different fixation methods and penetration time on the staining effect of human erythrocyte band 3 protein in immunofluorescence technique.Methods:Peripheral blood erythrocytes were collected from volunteers,and after cell smear or centrifugation,they were fixed using different fixatives(methanol or 2%paraformaldehyde-0.005%glutaraldehyde fixative),and after fixation,they were infiltrated by 0.1%Triton X-100 for 1-5 min,and the morphology of erythrocytes and the localization of band 3 proteins were observed using the immunofluorescence method.Results:Erythrocyte band 3 protein was expressed in the cell membrane.In the smear method of fixation,although the erythrocyte smear fixed by methanol caused a certain degree of cell shrinkage,the fixation effect was better and the loss of cells was less;the infiltration had little effect on the observation of its morphology and the localization of band 3 proteins;and the cells were seriously delaminated after fixing the coated erythrocytes using 2%paraformaldehyde-0.005%glutaraldehyde as the fixative solution.In the cell suspension fixation method,the erythrocyte suspension was fixed by 2%paraformaldehyde-0.005%glutaraldehyde,and the cells were hemolyzed and lost severely during the subsequent steps of centrifugal washing.The cell morphology and band 3 protein localization were best observed by 0.1%Triton X-100 infiltration for 1-3 min after cell smear fixation.After 5 min of infiltration,morphological changes of erythrocytes and fluorescence staining of cytoplasm were seen microscopically.Conclusion:In the immunofluorescence technique of peripheral blood erythrocytes,the band 3 proteins on the erythrocyte membrane can be better observed by using cell smear and methanol fixation for 1 min,and the action of 0.1%Triton X-100 for 1-3 min.关键词
免疫荧光技术/红细胞/带3蛋白/固定剂/渗透时间Key words
immunofluorescence technique/erythrocytes/band 3 protein/fixative/penetration time分类
基础医学引用本文复制引用
雷佳月,周永银,梁凯楹,刘志鑫,尹珂,林哲绚..免疫荧光技术中不同固定方式及渗透时间对人红细胞带3蛋白染色效果的影响[J].汕头大学医学院学报,2025,38(1):32-35,4.基金项目
广东省自然科学基金(2024A1515010890) （2024A1515010890）
广东省科技专项资金(210705156884432) （210705156884432）
