先进的 3D 高分辨率成像技术对于研究生物挑战至关重要，例如完整组织中的神经回路分析和肿瘤微环境。然而，澄清或扩增的生物样品中内源荧光蛋白发出的荧光信号随着重复照射和长时间成像而逐渐减弱，从而损害了其准确描述潜在科学问题的能力。我们开发了一种策略，可以在长时间的高分辨率三维成像过程中保留透明和扩展的组织样本中的荧光。我们评估了不同浓度的各种化合物，以确定它们增强荧光强度和抗光漂白性同时保持组织结构完整性的能力。具体来说，我们研究了 EDTP 利用对 GFP 的影响，因为已观察到它可以显着提高荧光强度、抗光漂白性，并在长期室温储存期间保持荧光。这一突破将促进扩展亲水性和基于水凝胶的透明和扩张方法，通过有效保护组织内的荧光蛋白来实现透明生物组织的长期高分辨率 3D 成像。© 2024。作者。

Advanced 3D high-resolution imaging techniques are essential for investigating biological challenges, such as neural circuit analysis and tumor microenvironment in intact tissues. However, the fluorescence signal emitted by endogenous fluorescent proteins in cleared or expanded biological samples gradually diminishes with repeated irradiation and prolonged imaging, compromising its ability to accurately depict the underlying scientific problem. We have developed a strategy to preserve fluorescence in cleared and expanded tissue samples during prolonged high-resolution three-dimensional imaging. We evaluated various compounds at different concentrations to determine their ability to enhance fluorescence intensity and resistance to photobleaching while maintaining the structural integrity of the tissue. Specifically, we investigated the impact of EDTP utilization on GFP, as it has been observed to significantly improve fluorescence intensity, resistance to photobleaching, and maintain fluorescence during extended room temperature storage. This breakthrough will facilitate extended hydrophilic and hydrogel-based clearing and expansion methods for achieving long-term high-resolution 3D imaging of cleared biological tissues by effectively safeguarding fluorescent proteins within the tissue.© 2024. The Author(s).