School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China.
School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China.
Sensors (Basel). 2023 Mar 21;23(6):3305. doi: 10.3390/s23063305.
Composed of a fluidic and an optical system, flow cytometry has been widely used for biosensing. The fluidic flow enables its automatic high-throughput sample loading and sorting while the optical system works for molecular detection by fluorescence for micron-level cells and particles. This technology is quite powerful and highly developed; however, it requires a sample in the form of a suspension and thus only works in vitro. In this study, we report a simple scheme to construct a flow cytometry based on a confocal microscope without any modifications. We demonstrate that line scanning of microscopy can effectively excite fluorescence of flowing microbeads or cells in a capillary tube in vitro and in blood vessels of live mice in vivo. This method can resolve microbeads at several microns and the results are comparable to a classic flow cytometer. The absolute diameter of flowing samples can be indicated directly. The sampling limitations and variations of this method is carefully analyzed. This scheme can be easily accomplished by any commercial confocal microscope systems, expands the function of them, and is of promising potential for simultaneous confocal microscopy and in vivo detection of cells in blood vessels of live animals by a single system.
流式细胞仪由流体系统和光学系统组成，已被广泛应用于生物传感领域。流体系统能够实现自动高通量的样品加载和分选，而光学系统则通过荧光对微米级别的细胞和颗粒进行分子检测。该技术非常强大且高度发达，但它需要以悬浮液的形式提供样品，因此仅在体外有效。在本研究中，我们报告了一种简单的方案，即在无需任何修改的情况下，基于共聚焦显微镜构建流式细胞仪。我们证明了显微镜的线扫描可以有效地激发体外毛细管中流动的微球或细胞的荧光，也可以在活体小鼠的血管内激发荧光。该方法可以分辨出几微米的微球，并且结果与经典的流式细胞仪相当。还可以直接指示流动样品的绝对直径。我们仔细分析了该方法的采样限制和变化。该方案可以通过任何商业共聚焦显微镜系统轻松实现，扩展了它们的功能，并且有望通过单个系统实现共聚焦显微镜和活体动物血管内细胞的同时检测。
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