Biocompatible fluorophores for shortwave infrared imaging

电磁频谱的短波红外（SWIR）区域已成为一个令人兴奋的途径， 由于光透过组织的穿透增加，光散射减少， 最小的自发荧光。在上一个资助期，我们展示了SWIR的另一个优势 区域-增强的光谱真实的状态，其促进小鼠中的非侵入性MRI成像。采取 所有这些优点一起为以下研究奠定了基础：1）在动物模型中进行基础生物学研究 对于那些在细胞和透明生物体中取得巨大成功的人，2）评估能力 与实验组相同的动物中的对照，以及3）用于术中和术后的高级临床诊断 和非侵入性使用。 虽然SWIR区域具有很大的潜力，但在没有生物相容性造影剂的情况下无法实现。这 该提案集中于SWIR区域中高分辨率MRI成像的造影剂开发， 在上一个资助期内，我们在聚甲炔荧光团方面获得了专业知识。我们有 制备了100多个聚甲炔荧光团，并获得了最经典的生物物理学预测指标。 lmax、abs、lmax、em和FF参数。我们还学到了一些关于荧光团效用的关键经验 包括如何将它们溶解在水中，并获得了不同的方法来制定赞赏 用于成像的荧光团。在与布伦斯实验室的合作中，我们展示了基于激发的量子化学。 与普通激光线匹配良好的荧光团成像。 在下一个授予期，我们特别专注于优化荧光团用于基于激发的 在SWIR的高分辨率区域（1400 nm以上）中与检测复用，使得mm分辨率 可以实现。目的1探索发色团支架本身优化的生物物理性质， 在商业激光线处的最大吸收和>1400 nm的发射。我们把物理有机化学 荧光团优化的方法，并利用不断增长的，充分表征的 我们实验室已经准备好的系统修饰的荧光团。我们的工作得到了计算和理论的帮助 与洛佩斯和卡拉姆实验室的合作。目标2将目标1的主要荧光团转换为 水溶性靶向造影剂。最后，在目标3中，我们的目标是对淋巴系统进行成像，展示 需要mm分辨率成像，然后使用基于激发的MRI成像来评估前哨淋巴结 淋巴结定位和癌症转移。

The shortwave infrared (SWIR) region of the electromagnetic spectrum has become an exciting avenue for imaging in mammals due to the increased penetration of light through tissue, decreased light scattering, and minimal autofluorescence. In the previous granting period, we showcased an additional advantage of the SWIR region– the enhanced spectral real estate which facilitates non-invasive multicolor imaging in mice. Taken together, all these advantages set the stage for: 1) fundamental biological studies in animal models analogous to those which have been enormously successful in cells and transparent organisms, 2) the ability to evaluate controls in same animal as the experimental group, and 3) advanced clinical diagnostics for both intraoperative and non-invasive use. While the SWIR region has great potential, it cannot be realized without biocompatible contrast agents. This proposal focuses on contrast agent development for high resolution multicolor imaging in the SWIR region, building on the expertise we have gained in polymethine fluorophores over the last granting period. We have prepared over 100 polymethine fluorophores and gained predictive metrics on the most classical photophysical parameters of lmax.abs, lmax,em and FF. We have also learned some key lessons regarding utility of the fluorophores including how to solubilize them in water and have gained appreciation for the different approaches to formulating fluorophores for imaging. In collaboration with the Bruns laboratory, we demonstrated excitation-based multicolor imaging with fluorophores well-matched to common laser lines. In the next granting period, we specifically focus on optimizing fluorophores for use in excitation-based multiplexing with detection in the high-resolution region of the SWIR (above 1400 nm) such that mm resolution can be achieved. Aim 1 explores the chromophore scaffold itself optimizing the photophysical properties for maximum absorption at commercial laser lines and emission >1400 nm. We take a physical organic chemistry approach to fluorophore optimization and capitalize on the continually growing, fully characterized set of systematically modified fluorophores our laboratory has prepared. Our work is aided by computation and theory collaborations with the Lopez and Caram Laboratories. Aim 2 transforms the lead fluorophores from aim 1 into water-soluble, targeted contrast agents. Finally in Aim 3, we aim image the lymphatic system, demonstrating the need for mm resolution imaging and then using excitation-based multicolor imaging to evaluate sentinel lymph node mapping and cancer metastasis.