Compact plane illumination plugin device to enable light sheet fluorescence imaging of multicellular organisms on an inverted wide-field microscope
Compact plane illumination plugin device to enable light sheet fluorescence imaging of multicellular organisms on an inverted wide-field microscope
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DOI：
10.1364/boe.7.000194
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发表时间：
2016-01-01
影响因子：
3.4
通讯作者：
Fei, Peng
中科院分区：
文献类型：
--
作者：
Guan, Zeyi;Lee, Juhyun;Fei, Peng
我们开发了一种紧凑型平面照明插件(PIP)装置，可以在传统倒置显微镜上实现平面照明和光片荧光成像。PIP设备允许将显微镜与可调激光片材轮廓、快速图像采集和三维扫描集成在一起。该设备既紧凑，尺寸约为15×5×5厘米，又具有成本效益，因为我们采用了消费电子产品和一种廉价的设备建模方法。我们通过成像不同的多细胞荧光结构，包括体外的3-D分支细胞和活的斑马鱼胚胎，证明了PIP提供了比传统显微镜显著的对比度和分辨率增强。与独立的倒置显微镜相比，集成了PIP的成像大大减少了焦外污染，并在采集的二维平面图像中产生了更清晰的对比度。从而对跳动的斑马鱼心脏的动态流体域进行了清晰的分割，实现了心脏的功能监测。此外，PIP的薄面照明提高了轴向分辨率，使得分枝细胞结构的三维重建成为可能，这导致了广视场显微镜功能的改进。(C)2015年美国光学学会
We developed a compact plane illumination plugin (PIP) device which enabled plane illumination and light sheet fluorescence imaging on a conventional inverted microscope. The PIP device allowed the integration of microscope with tunable laser sheet profile, fast image acquisition, and 3-D scanning. The device is both compact, measuring approximately 15 by 5 by 5 cm, and cost-effective, since we employed consumer electronics and an inexpensive device molding method. We demonstrated that PIP provided significant contrast and resolution enhancement to conventional microscopy through imaging different multi-cellular fluorescent structures, including 3-D branched cells in vitro and live zebrafish embryos. Imaging with the integration of PIP greatly reduced out-of-focus contamination and generated sharper contrast in acquired 2-D plane images when compared with the stand-alone inverted microscope. As a result, the dynamic fluid domain of the beating zebrafish heart was clearly segmented and the functional monitoring of the heart was achieved. Furthermore, the enhanced axial resolution established by thin plane illumination of PIP enabled the 3-D reconstruction of the branched cellular structures, which leads to the improvement on the functionality of the wide field microscopy. (C) 2015 Optical Society of America