E MAIL CONSULTATIONS

美国光生物学学会第26届年会 内源和给药荧光团的非线性激发 提供衍射受限的三维成像I细胞和 活组织制剂。多光子的主要优势 定量成像的激发是(1)固有的局部化 激发到一个小的明确的体积在一个高的焦点 数值孔径物镜，以及(2)激发能力 具有更好穿透性的紫外光和蓝光跃迁 近红外线照明。这些优势使高分辨率 荧光分子的成像，包括光敏化合物 例如Photofrin，在真实的肿瘤环境中。光敏蛋白 观察到激发过程从单光子过程变为双光子过程。 在700-740 nm范围内，在750-900范围内可以双光子激发 NM。它的特征是荧光截面为 在这个范围内，大约0.5×10-50cm4的S/光子。自.以来 源于NADH的线粒体自发荧光也经历了 双光子激发在这个波长范围内，两种物质都可以 同时成像。我们观察Photofrin最初定位到 质膜后的亚细胞定位 线粒体，可能还有其他细胞器。即使是在长时间 孵育，药物存在于组成肿瘤的细胞之间 椭球体。药物孵化导致方案依赖性减少 自体荧光。最后，观察到了光动力作用。 戏剧性地改变了细胞自发荧光的几个方面，如 以及细胞形态。在DRBIO由NIH RR04224支持。M.G.N。 是美国国立卫生研究院博士后研究员，由NRSA 1 F32 CA 72225-02资助。

26th annual meeting of the American Society for Photobiology Non-linear excitation of endogenous and administered fluorophores provides diffraction limited three-dimensional imaging I cells and living tissue preparations. The chief advantages of multiphoton excitation for quantitative imaging are (1) the inherent localization of excitation to a small well-defined volume at the focus of a high numerical aperture objective, and (2) the ability to excite ultraviolet and blue optical transitions with better penetrating near-infrared illumination. The advantages enable high-resolution imaging of fluorescent molecules, including photosensitizing compounds such as Photofrin, within realistic tumor environments. Photofrin excitation is observed to change from a one to two-photon process in the range from 700-740 nm and can be two-photon excited from 750-900 nm. It is characterized by a fluorescence cross section of approximately 0.5 x 10-50 cm4 s/photon throughout this range. Since mitochondrial autofluorescence originating from NADH also undergoes two-photon excitation in this wavelength range, both species can be simultaneously imaged. We observe Photofrin to localize initially to the plasma membrane followed by subcellular localization in mitochondria and possibly other organelles. Even after prolonged incubation, drug is present between cells comprising the tumor spheroids. Drug incubation results in protocol dependent decreases in autofluorescence. Finally, photodynamic action is observed to dramatically alter several aspects of cellular autofluorescence, as well as cell morphology. Supported at DRBIO by NIH RR04224. M.G.N. is a NIH postdoctoral fellow supported by NRSA 1 F32 CA 72225-02.