Zmijan Robert, Jonnalagadda Umesh S, Carugo Dario, Kochi Yu, Lemm Elizabeth, Packham Graham, Hill Martyn, Glynne-Jones Peter
Engineering Sciences, Faculty of Engineering and the Environment, University of Southampton, Southampton, SO17 1BJ, UK. Email:
Japan Patent Office, 3-chome-4-3 Kasumigaseki, Chiyoda-ku Tokyo 100-8915, Japan.
RSC Adv. 2015 Oct 31;5(101):83206-83216. doi: 10.1039/c5ra19497k. Epub 2015 Oct 1.
We demonstrate an imaging flow cytometer that uses acoustic levitation to assemble cells and other particles into a sheet structure. This technique enables a high resolution, low noise CMOS camera to capture images of thousands of cells with each frame. While ultrasonic focussing has previously been demonstrated for 1D cytometry systems, extending the technology to a planar, much higher throughput format and integrating imaging is non-trivial, and represents a significant jump forward in capability, leading to diagnostic possibilities not achievable with current systems. A galvo mirror is used to track the images of the moving cells permitting exposure times of 10 ms at frame rates of 50 fps with motion blur of only a few pixels. At 80 fps, we demonstrate a throughput of 208 000 beads per second. We investigate the factors affecting motion blur and throughput, and demonstrate the system with fluorescent beads, leukaemia cells and a chondrocyte cell line. Cells require more time to reach the acoustic focus than beads, resulting in lower throughputs; however a longer device would remove this constraint.
我们展示了一种成像流式细胞仪，它利用声悬浮将细胞和其他颗粒组装成片状结构。该技术使高分辨率、低噪声的互补金属氧化物半导体（CMOS）相机能够在每一帧中捕获数千个细胞的图像。虽然超声聚焦此前已在一维细胞计数系统中得到证明，但将该技术扩展到平面、高通量得多的形式并集成成像并非易事，这代表了能力上的重大飞跃，带来了当前系统无法实现的诊断可能性。使用一个振镜来跟踪移动细胞的图像，在帧率为50帧每秒时允许10毫秒的曝光时间，运动模糊仅为几个像素。在80帧每秒时，我们展示了每秒208000个珠子的通量。我们研究了影响运动模糊和通量的因素，并用荧光珠、白血病细胞和软骨细胞系演示了该系统。细胞比珠子需要更多时间到达声聚焦，导致通量较低；然而，更长的设备将消除这一限制。
