In vivo cell tracking with video rate multimodality laser scanning microscopy
In vivo cell tracking with video rate multimodality laser scanning microscopy
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DOI：
10.1109/jstqe.2007.912751
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发表时间：
2008-01-01
影响因子：
4.9
通讯作者：
Lin, Charles P.
中科院分区：
文献类型：
--
作者：
Veilleux, Israel;Spencer, Joel A.;Lin, Charles P.
生物过程的研究，如疾病进展和对治疗的反应，需要实时成像方法，允许连续观察，而不终止研究对象进行组织学组织处理。在所有当前的成像方式中，光学显微镜是唯一能够探测细胞和亚细胞分辨率的活组织的方法。我们提出了一个视频速率（30帧/秒），多模态成像系统，专为活体动物成像和细胞跟踪而设计。通过结合多种对比机制，包括反向散射、荧光（来自单光子和双光子激发）、二次谐波产生和相干反斯托克斯拉曼散射，利用共聚焦和非线性光学技术提取结构、功能和分子信息，获得体内深度剖分的高分辨率图像。同时使用多达三种模态是可能的，并且消除了对共配准的需要，特别是在大规模图像上。开发了一种实时运动校正算法，以延长积分时间，在情况下，图像需要稳定的对象运动。最后，对血管中快速移动的白细胞进行成像，可以在更小的区域内以120帧/秒的速度进行操作。用显微镜在体内获得的样品图像被提出来说明这种能力。
Studies of biological processes, such as disease progression and response to therapy, call for live imaging methods that allow continuous observation without terminating the study subject for histological tissue processing. Among all current imaging modalities, optical microscopy is the only method capable of probing live tissue with cellular and subcellular resolution. We present a video-rate (30 frames/s), multimodality imaging system that is designed specifically for live animal imaging and cell tracking. In vivo depth-sectioned, high-resolution images are obtained using confocal and nonlinear optical techniques that extract structural, functional, and molecular information by combining multiple contrast mechanisms, including back scattering, fluorescence (from single- and two-photon excitation), second harmonic generation, and coherent anti-Stokes Raman scattering. Simultaneous use of up to three modalities is possible and eliminates the need for coregistration, especially on large-scale images. A real-time movement correction algorithm was developed to extend integration times in cases where the image needs to be stabilized against subject movement. Finally, imaging of fast moving leukocytes in blood vessels is made possible with a modification that permits operation at 120 frames/s over a smaller area. Sample imagery obtained in vivo with the microscope is presented to illustrate the capabilities.