Gainer Christian F, Romanowski Marek
Dept. of Biomedical Engineering, University of Arizona, 1657 E Helen St, Tucson, AZ, USA 85721.
Proc SPIE Int Soc Opt Eng. 2013 Feb 22;8595:85950O-. doi: 10.1117/12.2002551.
While upconverting lanthanide nanoparticles have numerous advantages over other exogenous contrast agents used in scanned multiphoton imaging, their long luminescence lifetimes cause images collected with non-descanned detection to be greatly blurred. We demonstrate herein the use of Richardson-Lucy deconvolution to deblur luminescence images obtained via multiphoton scanning microscopy. Images were taken of three dimensional models of colon and ovarian cancer following incubation with NaYF:Yb,Er nanoparticles functionalized with an antibody for EGFR and folic acid respectively. Following deconvolution, images had a lateral resolution on par with the optimal performance of the imaging system used, ~1.2 μm, and an axial resolution of ~5 μm. Due to the relatively high multiphoton excitation efficiency of these nanoparticles, it is possible to follow binding of individual particles in tissue. In addition, their extreme photostability allows for prolonged imaging without significant loss in luminescence signal. With these advantageous properties in mind, we also discuss the potential application of upconverting lanthanide nanoparticles for tracking of specific, cancer relevant receptors in tissue.
虽然上转换镧系纳米粒子相较于扫描多光子成像中使用的其他外源性造影剂具有众多优势，但其较长的发光寿命会导致采用非反扫描检测收集的图像严重模糊。我们在此展示了使用理查森 - Lucy反卷积来对通过多光子扫描显微镜获得的发光图像进行去模糊处理。在用分别用针对表皮生长因子受体（EGFR）的抗体和叶酸功能化的NaYF:Yb,Er纳米粒子孵育后，对结肠癌和卵巢癌的三维模型进行了成像。反卷积后，图像的横向分辨率与所使用成像系统的最佳性能相当，约为1.2μm，轴向分辨率约为5μm。由于这些纳米粒子相对较高的多光子激发效率，有可能追踪组织中单个粒子的结合情况。此外，它们极高的光稳定性使得能够进行长时间成像而发光信号无显著损失。考虑到这些有利特性，我们还讨论了上转换镧系纳米粒子在追踪组织中特定的、与癌症相关的受体方面的潜在应用。
