Lin Meiai, Qiao Xu, Liu Qiao, Shao Changshun, Su Xuantao
Institute of Biomedical Engineering, School of Control Science and Engineering, Shandong University, Jinan, Shandong, 250061, China.
Key Laboratory of Experimental Teratology (Ministry of Education); Department of Molecular Medicine and Genetics, School of Medicine, Shandong University, Jinan, Shandong, 250012, China.
Biomed Opt Express. 2016 Nov 16;7(12):5170-5181. doi: 10.1364/BOE.7.005170. eCollection 2016 Dec 1.
A light-sheet-based 2D light scattering cytometer is developed for label-free characterization of senescent cells. The light-sheet provides an illumination beam with controlled thickness for single cell excitation, and 2D light scattering patterns are obtained by using a defocused imaging method. The principle of this cytometer is validated by distinguishing microspheres with submicron resolution. Automatic classification of senescent and normal cells is achieved at single cell level by using the support vector machine (SVM) algorithm, where a sensitivity of 89.1% and a specificity of 96.4% are obtained. Our results suggest that the light-sheet-based 2D light scattering label-free cytometry has the capability to perform size differentiation of beads with submicron resolution and to classify different groups of cells without fluorescent labeling, showing the potential for clinical diagnosis of senescence-related diseases.
一种基于光片的二维光散射细胞仪被开发用于对衰老细胞进行无标记表征。光片提供具有可控厚度的照明光束用于单细胞激发,并且通过使用散焦成像方法获得二维光散射图案。通过以亚微米分辨率区分微球验证了该细胞仪的原理。使用支持向量机(SVM)算法在单细胞水平实现了衰老细胞和正常细胞的自动分类,其中灵敏度为89.1%,特异性为96.4%。我们的结果表明,基于光片的二维光散射无标记细胞术有能力以亚微米分辨率对珠子进行尺寸区分,并对不同组的细胞进行无荧光标记分类,显示出在衰老相关疾病临床诊断中的潜力。