Characterization of flow direction in microchannels and zebrafish blood vessels by scanning fluorescence correlation spectroscopy
Characterization of flow direction in microchannels and zebrafish blood vessels by scanning fluorescence correlation spectroscopy
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DOI：
10.1117/1.2435173
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发表时间：
2007-01-01
影响因子：
3.5
通讯作者：
Wohland, Thorsten
中科院分区：
文献类型：
--
作者：
Pan, Xiaotao;Yu, Hanry;Wohland, Thorsten
利用荧光相关光谱技术研究微结构和生物组织中的流体流动是近十年来一个具有挑战性的课题。由于其固有的光学配置，圆形聚焦激光束，FCS无法解决微流体流动方向。早期的方案报道了使用两个激光束或使用非对称激光焦点来打破测量系统的对称性。然而，这难以与共焦系统联合收割机组合，因为它将需要干扰成像能力的修改。我们提出了一种称为线扫描FCS的方法来测量微通道和组织中的不同流动角度。该方法在组合的激光扫描共聚焦显微镜（LSCM）和FCS系统上实现，该系统能够实现不妥协的成像和光谱测量。我们证明，通过扫描激光束与一个定义的速度和方向，我们可以测量流动方向与当前的系统在至少3 μ m的最佳分辨率。该组合系统通过测量在有和没有阻塞的微通道中的流动剖面来评估。为了将该技术扩展到活组织测量，我们证明了线扫描FCS可以在无标记的方法中确定斑马鱼小血管中的流动方向。(c)2007年，由光学仪器工程师协会（Society of Photo-Optical Instrumentation Engineers）主办。
The investigation of flow profiles in microstructures and tissues by fluorescence correlation spectroscopy (FCS) has been a challenging topic in the past decade. Due to its inherent optical configuration, a circular focused laser beam, FCS is unable to resolve microfluidic flow directions. Earlier schemes reported the use of two laser beams or the use of nonsymmetrical laser foci to break the symmetry of the measurement system. This, however, is difficult to combine with confocal systems since it would require modifications that interfere with the imaging capabilities. We propose a method called linescan FCS to measure different flow angles in microchannels and tissues. This method is implemented on a combined laser scanning confocal microscopy (LSCM) and FCS system that enables uncompromised imaging and spectroscopy measurements. We demonstrate that by scanning the laser beam with a defined speed and direction we can measure flow direction with the current system at an optimal resolution of at least 3 mu m. The combination system is assessed by measuring flow profiles in a microchannel with and without obstruction. To extend the technique to live tissue measurements we demonstrate that line-scan FCS can determine the flow direction in zebrafish small blood vessels in a label-free approach. (c) 2007 Society of Photo-Optical Instrumentation Engineers.