CNR - ISASI, Istituto di Scienze Applicate e Sistemi Intelligenti "E. Caianiello", Via Campi Flegrei 34, 80078 Pozzuoli, NA, Italy.
CNR - ISASI, Istituto di Scienze Applicate e Sistemi Intelligenti "E. Caianiello", Via Campi Flegrei 34, 80078 Pozzuoli, NA, Italy.
Methods. 2018 Mar 1;136:108-115. doi: 10.1016/j.ymeth.2018.01.003. Epub 2018 Jan 16.
High-throughput single-cell analysis is a challenging target for implementing advanced biomedical applications. An excellent candidate for this aim is label-free tomographic phase microscopy (TPM). In this paper, some of the methods used to obtain TPM are reviewed, analyzing advantages and disadvantages of each of them. Moreover, an alternative tomographic technique is described for live cells analysis, and future trends of the method are foreseen. In particular, by exploiting random rolling of cells while they are flowing along a microfluidic channel, it is possible to obtain phase-contrast tomography thus obtaining complete retrieval of both 3D-position and orientation of rotating cells. Thus, a priori knowledge of such information is no longer needed. This approach extremely simplifies the optical system avoiding any mechanical/optical scanning of light source. The proof is given for different classes of biosamples, red-blood-cells (RBCs) and diatom algae. Accurate characterization of each type of cells is reported and compared to that obtained by other tomographic techniques.
高通量单细胞分析是实现先进生物医学应用的挑战性目标。无标记断层相显微镜(TPM)是实现这一目标的优秀候选者。本文回顾了获得 TPM 的一些方法,分析了它们各自的优缺点。此外,还描述了一种用于活细胞分析的替代断层技术,并预测了该方法的未来趋势。特别是,通过利用细胞在微流道中流动时的随机滚动,可以获得相衬断层成像,从而完全恢复旋转细胞的三维位置和方向。因此,不再需要这种信息的先验知识。这种方法极大地简化了光学系统,避免了光源的任何机械/光学扫描。本文针对不同类型的生物样本,即红细胞(RBC)和硅藻,给出了证明。报告了对每种类型细胞的精确特征,并与其他断层技术获得的结果进行了比较。