Multiphoton autofluorescence imaging of intratissue elastic fibers
Multiphoton autofluorescence imaging of intratissue elastic fibers
复制标题
DOI：
10.1016/j.biomaterials.2004.02.059
复制
发表时间：
2005-02-01
期刊：
影响因子：
14
通讯作者：
Stock, UA
中科院分区：
文献类型：
--
作者：
König, K;Schenke-Layland, K;Stock, UA
通过近红外飞秒激光脉冲的多光子诱导的蓝色/绿色自体荧光已被用于选择性地对天然和组织工程（TE）存活心脏瓣膜中的组织内弹性纤维进行成像，而无需任何侵入性组织去除、包埋、固定和染色。弹性纤维可以清楚地与胶原结构区分开，胶原结构在用由于二次谐波产生的强超短脉冲激发时发射紫外/紫色辐射。通过使用多光子激光扫描显微镜与可调谐80 MHz飞秒激光源连接的心脏瓣膜的光学切片，进行具有亚微米空间分辨率的弹性纤维的深层组织三维成像。该技术用于在植入前诊断TE心脏瓣膜的细胞外基质结构和细胞重新安置。这种新的非侵入性方法开启了在不染色的情况下对活的完整组织中的弹性纤维、胶原结构和细胞内细胞器进行高分辨率原位成像的一般可能性（C）2004 Elsevier Ltd.保留所有权利。
Multiphoton induced blue/green autofluorescence by near infrared femtosecond laser pulses has been used to selectively image intratissue elastic fibers in native and tissue engineered (TE) viable heart valves without any invasive tissue removal, embedding, fixation, and staining. Elastic fibers could be clearly distinguished from collagenous Structures which emit ultraviolet/violet radiation when excited with intense ultrashort Pulses due to second harmonic generation. Deep-tissue three-dimensional imaging of elastic fibers with submicron spatial resolution was performed by optical sectioning of heart valves using a multiphoton laser scanning microscope in connection with a tunable 80 MHz femtosecond laser source. The technology was used to diagnose extracellular matrix structures and cell resettlement of TE heart valves prior implantation. This novel non-invasive method opens the general possibility of high- resolution in situ imaging of elastic fibers, collagen structures and intracellular organelles in living intact tissues without staining (C) 2004 Elsevier Ltd. All rights reserved.