Electrical Engineering Department, University of California, Los Angeles, CA 90095, USA.
Analyst. 2011 Sep 7;136(17):3512-8. doi: 10.1039/c0an00926a. Epub 2011 Jan 31.
We demonstrate lensless fluorescent microscopy over a large field-of-view of ~60 mm(2) with a spatial resolution of <4 µm. In this on-chip fluorescent imaging modality, the samples are placed on a fiber-optic faceplate that is tapered such that the density of the fiber-optic waveguides on the top facet is >5 fold larger than the bottom one. Placed on this tapered faceplate, the fluorescent samples are pumped from the side through a glass hemisphere interface. After excitation of the samples, the pump light is rejected through total internal reflection that occurs at the bottom facet of the sample substrate. The fluorescent emission from the sample is then collected by the smaller end of the tapered faceplate and is delivered to an opto-electronic sensor-array to be digitally sampled. Using a compressive sampling algorithm, we decode these raw lensfree images to validate the resolution (<4 µm) of this on-chip fluorescent imaging platform using microparticles as well as labeled Giardia muris cysts. This wide-field lensfree fluorescent microscopy platform, being compact and high-throughput, might provide a valuable tool especially for cytometry, rare cell analysis (involving large area microfluidic systems) as well as for microarray imaging applications.
我们展示了一种无透镜荧光显微镜,其视场约为 60mm²,空间分辨率小于 4µm。在这种片上荧光成像模式中,样品放置在光纤面板上,该光纤面板逐渐变细,使得上表面的光纤波导密度比下表面大 5 倍以上。将荧光样品放置在这个锥形的光纤面板上,通过玻璃半球形界面从侧面泵浦。在激发样品后,通过在样品基底的下表面发生全内反射来拒绝泵浦光。然后,通过锥形光纤面板的较小端收集来自样品的荧光发射,并将其传输到光电传感器阵列进行数字采样。使用压缩采样算法,我们对这些原始无透镜图像进行解码,以验证使用微粒子以及标记的鼠贾第虫囊包作为模型的这种片上荧光成像平台的分辨率(<4µm)。这种宽视场无透镜荧光显微镜平台具有紧凑和高通量的特点,可能为细胞术、稀有细胞分析(涉及大面积微流控系统)以及微阵列成像应用提供一种有价值的工具。