Current circulating tumor cells (CTC) detection methods have to compromise between sensitivity and throughput. High-throughput imaging cytometer based on serial time-encoded amplified microscopy (STEAM) facilitates CTC detection at single-cell sensitivity from abundant cells. However, this method lacks the information to spot heterogeneity of cells with high morphological similarity. Researches on cell biophysical properties suggest cell mechanotyping can be an indicator of phenotypic heterogeneity to improve classification ability of STEAM cytometer. Here, we present a high-throughput label-free acoustofluidic imaging cytometer for single-cell mechanotyping based on STEAM and acoustofluidic technology. The generated acoustic resonance field translocates cells to different transversal exit positions under continuous flow according to their intrinsic biophysical properties. Such displacements are recorded with images simultaneously using STEAM cytometry at approximately 2000 cells/s. We experimentally verified that our method accounting for both cell images and acoustic displacements can improve mechanotyping accuracy by 12% upon image-based phenotyping method. This new acoustofluidic imaging cytometer facilitates high-accuracy and high-throughput imaging cytometry for single-cell CTC mechanotyping.

当前的循环肿瘤细胞（CTC）检测方法必须在灵敏度和通量之间折衷。基于串行时间编码放大显微镜（STEAM）的高通量成像细胞仪有助于在大量细胞的单细胞敏感性下进行CTC检测。但是，这种方法缺乏信息来发现具有高度形态相似性的细胞异质性。细胞生物学特性的研究表明，细胞机械分型可以作为表型异质性的指标，以提高STEAM细胞计数仪的分类能力。在这里，我们提出了一种基于STEAM和声流技术的高通量无标记声流成像细胞仪，用于单细胞机械分型。产生的声共振场根据其固有的生物物理特性，在连续流动下将细胞移位到不同的横向出口位置。使用STEAM细胞计数仪同时以大约2000个细胞/秒的速度同时记录此类位移和图像。我们通过实验验证了我们的方法同时考虑了细胞图像和声学位移，与基于图像的表型方法相比，可以将机械分型精度提高12％。这款新型的声流成像细胞仪可促进用于单细胞CTC机械分型的高精度和高通量成像细胞仪。我们通过实验验证了我们的方法同时考虑了细胞图像和声学位移，与基于图像的表型方法相比，可以将机械分型精度提高12％。这款新型的声流成像细胞仪可促进用于单细胞CTC机械分型的高精度和高通量成像细胞仪。我们通过实验验证了我们的方法同时考虑了细胞图像和声学位移，与基于图像的表型方法相比，可以将机械分型精度提高12％。这款新型的声流成像细胞仪可促进用于单细胞CTC机械分型的高精度和高通量成像细胞仪。