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Giga-pixel fluorescent imaging over an ultra-large field-of-view using a flatbed scanner

机译:使用平板扫描仪在超大视野内进行千兆像素荧光成像

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摘要

We demonstrate a new fluorescent imaging technique that can screen for fluorescent micro-objects over an ultra-wide field-of-view (FOV) of ~532 cm2, i.e., 19 cm × 28 cm, reaching a space-bandwidth product of more than 2 billion. For achieving such a large FOV, we modified the hardware and software of a commercially available flatbed scanner, and added a custom-designed absorbing fluorescent filter, a two-dimensional array of external light sources for computer-controlled and high-angle fluorescent excitation. We also re-programmed the driver of the scanner to take full control of the scanner hardware and achieve the highest possible exposure time, gain and sensitivity for detection of fluorescent micro-objects through the gradient index self-focusing lens array that is positioned in front of the scanner sensor chip. For example, this large FOV of our imaging platform allows us to screen more than 2.2 mL of undiluted whole blood for detection of fluorescent micro-objects within <5 minutes. This high-throughput fluorescent imaging platform could be useful for rare cell research and cytometry applications by enabling rapid screening of large volumes of optically dense media. Our results constitute the first time that a flatbed scanner has been converted to a fluorescent imaging system, achieving a record large FOV.
机译:我们演示了一种新的荧光成像技术,该技术可以在约532 cm 2 (即19 cm×28 cm)的超宽视场(FOV)上筛选荧光微对象超过20亿的空间带宽产品。为了实现如此大的FOV,我们修改了市售平板扫描仪的硬件和软件,并添加了定制设计的吸收式荧光滤光片,用于计算机控制和高角度荧光激发的二维外部光源阵列。我们还对扫描仪的驱动程序进行了重新编程,以完全控制扫描仪硬件,并通过位于前面的梯度折射率自聚焦透镜阵列实现了最大可能的曝光时间,增益和灵敏度,以检测荧光微物体。扫描仪传感器芯片。例如,我们成像平台的这种大视野使我们能够在不到5分钟的时间内筛选出2.2 mL以上的未稀释全血,以检测荧光微物体。这种高通量荧光成像平台可通过快速筛选大量的光致密性培养基,对稀有细胞研究和细胞计数应用有用。我们的结果是平板扫描仪首次转换为荧光成像系统,实现了创纪录的大视野。

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