Objective : To alleviate the severe limitation of the prohibitively long process of immune-fluorescence labeling on the routine applications of revolutionary intact tissue clearing techniques in diverse biomedical arenas. Methods : We proposed an easily adaptable approach, electro-enhanced rapid staining (EERS), for highly efficient and fast immuno-labeling of thick clarified tissues. In EERS, an optimized and precisely controlled weak external electric field is engineered into a compact device to enable efficient and uniform transport of antibodies into clarified tissues while minimizing the detrimental effect of macromolecular crowding at the tissue-solution interface. Results and Conclusions : The experimental results show that, with EERS, a current density of only ∼0.2 mA mm ⁻² is sufficient to achieve uniform labeling of clarified tissues of several millimeters thick in a few hours without detectable tissue damage. In addition, the amount of antibodies required is also several-fold lower than conventional immuno-labeling assays under comparable conditions. Significance : It is expected that the implementation of EERS in most laboratories should significantly expedite the application of tissue clearing in a broad range of research explorations, both basic and clinical.

中文翻译：

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使用低场增强扩散对澄清组织进行高效快速的免疫标记

目的：减轻免疫荧光标记的漫长过程对革命性完整组织清除技术在不同生物医学领域的常规应用的严重限制。 方法：我们提出了一种易于适应的方法，即电增强快速染色 (EERS)，用于对厚澄清组织进行高效和快速的免疫标记。在 EERS 中，优化和精确控制的弱外部电场被设计成一个紧凑的设备，使抗体能够高效、均匀地传输到澄清的组织中，同时最大限度地减少组织-溶液界面处大分子拥挤的不利影响。结果和结论：实验结果表明，使用 EERS，仅 ~0.2 mA mm ^-2的电流密度 就足以在几小时内对几毫米厚的澄清组织进行均匀标记，而不会检测到组织损伤。此外，在可比较的条件下，所需的抗体量也比传统的免疫标记分析低几倍。意义：预计在大多数实验室实施 EERS 将显着加快组织清除在广泛的基础和临床研究探索中的应用。