Dannhauser David, Rossi Domenico, Memmolo Pasquale, Finizio Andrea, Ferraro Pietro, Netti Paolo Antonio, Causa Filippo
Center for Advanced Biomaterials for Health Care@CRIB, Istituto Italiano di Tecnologia, Largo Barsanti e Matteucci 53, 80125 Naples, Italy.
CNR-ISASI Institute of Applied Sciences & Intelligent Systems "E. Caianiello", Via Campi Flegrei 34, 80078 Pozzuoli, Italy.
Biomed Opt Express. 2018 Oct 4;9(11):5194-5204. doi: 10.1364/BOE.9.005194. eCollection 2018 Nov 1.
We implemented a completely label-free biophysical (morphometric and optical) property characterization of living monocytes in flow, using measurements obtained from two coherent imaging techniques: a pure light scattering approach to obtain an optical signature (OS) of cells, and a digital holography (DH) approach to achieve optical cell reconstructions in flow. A precise 3D cell alignment platform, taking advantage of viscoelastic fluid properties and microfluidic channel geometry, was used to investigate the OS of cells to achieve their refractive index, ratio of the nucleus over cytoplasm, and overall cell dimension. Further quantitative phase-contrast reconstructions by DH were employed to calculate surface area, dry mass, and biovolume of monocytes by using the OS outcomes as input parameters. The results show significantly different biophysical cell properties, confirming the possibility to differentiate monocytes from other cell classes in flow, thus avoiding chemical cell staining or labeling, which are nowadays used.
我们采用两种相干成像技术获得的测量结果，对流动状态下的活单核细胞进行了完全无标记的生物物理（形态测量和光学）特性表征：一种是通过纯光散射方法获取细胞的光学特征（OS），另一种是利用数字全息术（DH）方法在流动状态下实现细胞的光学重建。利用粘弹性流体特性和微流控通道几何形状的精确三维细胞对齐平台，研究细胞的光学特征，以获得其折射率、细胞核与细胞质的比例以及细胞整体尺寸。通过数字全息术进行的进一步定量相衬重建，以光学特征结果作为输入参数，用于计算单核细胞的表面积、干质量和生物体积。结果显示出明显不同的生物物理细胞特性，证实了在流动状态下区分单核细胞与其他细胞类型的可能性，从而避免了如今所使用的化学细胞染色或标记。
