Jaiswal Devina, Moscato Zoe, Tomizawa Yuji, Claffey Kevin P, Hoshino Kazunori
Department of Biomedical Engineering, University of Connecticut, 260 Glenbrook Rd, Storrs, Connecticut 06269, USA.
Department of Biomedical Engineering, Western New England University, 1215 Wilbraham Rd, Springfield, Massachusetts 01119, USA.
Biomed Opt Express. 2019 Apr 15;10(5):2409-2418. doi: 10.1364/BOE.10.002409. eCollection 2019 May 1.
We have demonstrated a new method of 3D elastography based on 3D light microscopy and micro-scale manipulation. We used custom-built micromanipulators to apply a mechanical force onto multicellular tumor spheroids (200-300 µm in size) and recorded the induced compression with a differential interference contrast (DIC)/confocal microscope to obtain a 4D (x, y, z, and indentation steps) image sequence. Deformation analysis made through 3D pattern tracking without using fluorescence revealed 3D structural and spatial heterogeneity in tumor spheroids. We observed a 20-30 µm-sized spot of locally-induced large deformation within a tumor spheroid. We also found solid fibroblast cores formed in a tumor-fibroblast co-culture spheroid to be stiffer than surrounding cancer cells, which would not have been discovered using only conventional fluorescence. Our new method of 3D elastography may be used to better understand structural composition in multicellular spheroids through analysis of mechanical heterogeneity.
我们展示了一种基于三维光学显微镜和微观尺度操作的新型三维弹性成像方法。我们使用定制的微操纵器对多细胞肿瘤球体（尺寸为200 - 300微米）施加机械力，并用微分干涉对比（DIC）/共聚焦显微镜记录诱导的压缩情况，以获得四维（x、y、z和压痕步骤）图像序列。通过不使用荧光的三维模式跟踪进行的变形分析揭示了肿瘤球体中的三维结构和空间异质性。我们在肿瘤球体中观察到一个尺寸为20 - 30微米的局部诱导大变形点。我们还发现，在肿瘤 - 成纤维细胞共培养球体中形成的实性成纤维细胞核心比周围癌细胞更硬，这是仅使用传统荧光方法无法发现的。我们的新型三维弹性成像方法可通过分析机械异质性来更好地理解多细胞球体中的结构组成。
