Vajpayee Srishti, Picascia Tiziana, Casciano Fabio, Viale Elisabetta, Ronda Luca, Bettati Stefano, Milani Daniela, Gretz Norbert, Perciaccante Rossana
Cyanagen Srl, Via degli Stradelli Guelfi 40/C, 40138 Bologna, Italy.
Medical Research Center, Medical Faculty Mannheim, University of Heidelberg, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Germany.
Chem Biomed Imaging. 2024 Sep 10;2(10):721-730. doi: 10.1021/cbmi.4c00028. eCollection 2024 Oct 28.
Over the last decades, various tissue-clearing techniques have broken the ground for the optical imaging of whole organs and whole-organisms, providing complete representative data sets of three-dimensional biological structures. Along with advancements in this field, the development of fluorescent markers for staining vessels and capillaries has offered insights into the complexity of vascular networks and their impact on disease progression. Here we describe the use of a modified water-soluble chitosan linked to cyanine dyes in combination with ethyl cinnamate-based optical tissue clearing for the 3D visualization of tissue vasculature in depth. The water-soluble fluorescent Chitosans have proven to be an optimal candidate for labeling both vessels and capillaries thanks to their increased water solubility, high photostability, and optical properties in the near-infrared window. In addition, the nontoxicity of these markers broadens their applicability to and biological applications. Despite the availability of other fluorescent molecules for vascular staining, the present study, for the first time, demonstrates the potential of fluorescent chitosans to depict vessels at the capillary level and opens avenues for advancing the diagnostic field by reducing the complexity and costs of the currently available procedures.
在过去几十年中，各种组织透明化技术为全器官和全生物体的光学成像开辟了道路，提供了三维生物结构的完整代表性数据集。随着该领域的进展，用于血管和毛细血管染色的荧光标记物的开发为深入了解血管网络的复杂性及其对疾病进展的影响提供了帮助。在此，我们描述了一种与花青染料连接的改性水溶性壳聚糖与基于肉桂酸乙酯的光学组织透明化技术相结合，用于深度三维可视化组织脉管系统。水溶性荧光壳聚糖因其增加的水溶性、高光稳定性以及在近红外窗口的光学特性，已被证明是标记血管和毛细血管的理想候选物。此外，这些标记物的无毒性拓宽了它们在生物医学和生物学应用中的适用性。尽管有其他用于血管染色的荧光分子，但本研究首次证明了荧光壳聚糖在描绘毛细血管水平血管方面的潜力，并通过降低现有程序的复杂性和成本为推进诊断领域开辟了道路。
