Li Chunqiang, Pastila Riikka K, Pitsillides Costas, Runnels Judith M, Puoris'haag Mehron, Côté Daniel, Lin Charles P
Wellman Center for Photomedicine and Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Opt Express. 2010 Jan 18;18(2):988-99. doi: 10.1364/OE.18.000988.
We describe a new method for imaging leukocytes in vivo by exciting the endogenous protein fluorescence in the ultraviolet (UV) spectral region where tryptophan is the major fluorophore. Two-photon excitation near 590 nm allows noninvasive optical sectioning through the epidermal cell layers into the dermis of mouse skin, where leukocytes can be observed by video-rate microscopy to interact dynamically with the dermal vascular endothelium. Inflammation significantly enhances leukocyte rolling, adhesion, and tissue infiltration. After exiting the vasculature, leukocytes continue to move actively in tissue as observed by time-lapse microscopy, and are distinguishable from resident autofluorescent cells that are not motile. Because the new method alleviates the need to introduce exogenous labels, it is potentially applicable for tracking leukocytes and monitoring inflammatory cellular reactions in humans.
我们描述了一种通过激发紫外线（UV）光谱区域内的内源性蛋白质荧光来对体内白细胞进行成像的新方法，在该区域色氨酸是主要的荧光团。590nm附近的双光子激发允许通过表皮细胞层对小鼠皮肤真皮进行非侵入性光学切片，在真皮中可以通过视频速率显微镜观察到白细胞与真皮血管内皮细胞动态相互作用。炎症显著增强白细胞滚动、黏附和组织浸润。通过延时显微镜观察，白细胞从脉管系统出来后，在组织中继续活跃移动，并且可与不移动的驻留自发荧光细胞区分开来。由于这种新方法无需引入外源性标记，因此有可能适用于追踪人类白细胞和监测炎症细胞反应。
