Abstract：
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.
摘要：
目的探讨一种简单可行的ApoE-/-小鼠动脉粥样硬化模型淋巴管整体免疫荧光染色方法。方法从ApoE-/-小鼠模型中仔细切除主动脉标本。用针对平滑肌肌动蛋白（SMA）和淋巴管内皮受体1（LYVE1）的特异性抗体进行免疫染色后，主动脉,包括主动脉根部,用5g/L苏丹黑B溶液进行30分钟处理。该步骤有助于最小化组织的自发荧光背景。此后,通过清除方案处理主动脉,并在荧光显微镜下在特制的腔室内成像.结果5g/L苏丹黑B预处理可有效抑制血管结构发出的自发荧光信号。从而增强与淋巴管相关的特定荧光信号的对比度和清晰度。信号质量的这种增强不会损害免疫荧光标记的完整性或特异性。结论一个简单的，高度特异性,建立了ApoE-/-小鼠整装主动脉制剂中淋巴管可视化的有效方法。
