载脂蛋白E敲除(ApoE^(-/-))小鼠整装(whole-mount)主动脉免疫荧光组织化学染色的血管外膜淋巴管成像  

Imaging for displaying lymphatic vessels in whole-mount aorta adventitia of ApoE^(-/-) mice by immunofluorescent histochemical staining

作　　者：何玉莲[1] 钟文飞 林彩燕[1] 李玉婷[1] 冯森玲 严鹏科[1] HE Yulian;ZHONG Wenfei;LIN Caiyan;LI Yuting;FENG Senling;YAN Pengke(Department of Pharmacy,Guangdong Provincial Key Laboratory of Major Obstetric Diseases,Guangdong Provincial Clinical Research Center for Obstetrics and Gynecology,The Third Affiliated Hospital of Guangzhou Medical University,Guangzhou 510150;Guangzhou Panyu District Shibi Street Community Health Center,Guangzhou 511400,China)

机构地区：[1]药学部,广东省产科重大疾病重点实验室,广东省妇产疾病临床医学研究中心,广州医科大学附属第三医院,广东广州510150 [2]广州市番禺区石壁街社区卫生服务中心,广东广州511400

出　　处：《细胞与分子免疫学杂志》2024年第6期527-531,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：广东省自然科学基金(2022A1515010199);毅文人才工程启航计划([2021]09);贺林院士新医学科研基金(2021HLKY02)。

摘　　要：目的 探索载脂蛋白E敲除(ApoE^(-/-))小鼠整装(whole-mount)主动脉免疫荧光组织化学染色法显示血管外膜淋巴管的可行性。方法 制备ApoE^(-/-)小鼠整装主动脉,进行淋巴管内皮受体1(LYVE1)、 α平滑肌肌动蛋白(α-SMA)免疫荧光组织化学染色,染色后组织经5 g/L苏丹黑B处理30 min、组织透明液透明,移至自制的样本容器中,荧光显微镜下成像。结果经5 g/L苏丹黑B处理的整装主动脉可见血管自发荧光显著降低,外膜淋巴管的特异性荧光强度显著增强,更易观察且不会对其他通道荧光产生干扰。结论 建立的ApoE^(-/-)小鼠整装主动脉免疫荧光组织化学染色显示血管外膜淋巴管的方法操作简便,特异性好。Objective To explore a simple and feasible method for whole-mount immunofluorescence staining of lymphatic vessels in the ApoE^(-/-)mouse model of atherosclerosis.Methods Aortic specimens were carefully excised from the ApoE^(-/-)mouse model.Following immunostaining with specific antibodies against smooth muscle actin(SMA)and lymphatic vessel endothelial receptor 1(LYVE1),the aortas,including the aortic root,were subjected to a 30-minute treatment with 5 g/L Sudan Black B solution.This step was instrumental in minimizing the autofluorescent background of the tissue.Thereafter,the aortas were processed through a clearing protocol and imaged within a purpose-built chamber under a fluorescence microscope.Results The pretreatment with 5 g/L Sudan Black B effectively suppressed the autofluorescent signals emanating from the vascular structures,thereby enhancing the contrast and clarity of the specific fluorescence signals associated with the lymphatic vessels.This enhancement in signal quality did not compromise the integrity or specificity of the immunofluorescent markers.Conclusion A facile,highly specific,and effective approach for the visualization of lymphatic vessels in whole-mount aortic preparations from ApoE^(-/-)mice is established.

关 键 词：载脂蛋白E敲除(ApoE^(-/-)) 整装(whole-mount)小鼠主动脉 动脉粥样硬化 淋巴管 苏丹黑B 

分 类 号：R543.5[医药卫生—心血管疾病] R322.26[医药卫生—内科学] R543.1[医药卫生—临床医学] TS193.5[轻工技术与工程—纺织化学与染整工程]