AFM stiffness nanotomography of normal, metaplastic and dysplastic human esophageal cells.
AFM stiffness nanotomography of normal, metaplastic and dysplastic human esophageal cells.
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正常，化生和发育不良的人类食管细胞的AFM刚度纳米术。
DOI：
10.1088/1478-3975/8/1/015007
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发表时间：
2011-02
期刊：
影响因子：
2
通讯作者：
Ros R
中科院分区：
文献类型：
--
作者：
Fuhrmann A;Staunton JR;Nandakumar V;Banyai N;Davies PC;Ros R
单个细胞的机械刚度对于组织稳态、细胞生长、分裂和运动以及癌症发生过程中的上皮-间质转化非常重要。在这项工作中，研究了正常鳞状细胞系 (EPC2) 和化生细胞系 (CP-A) 以及发育不良 (CP-D) 巴雷特食管柱状细胞系，作为人类食管肿瘤前期进展的模型。我们结合使用原子力显微镜（AFM）和扫描共焦荧光寿命成像显微镜（FLIM）来研究单个贴壁细胞的机械特性。以 8×8 网格图案对每个细胞的细胞核绘制 64 条力压痕曲线。分析力压痕曲线，发现所有细胞系都具有与压痕深度相关的杨氏模量。刚度断层扫描显示所研究细胞系的机械性能之间存在明显差异。比较 1 nN 压痕力的刚度，计算出 EPC2（n=18 个细胞）最可能的杨氏模量为 4.7 kPa，CP-A（n=10）为 3.1 kPa，CP-D（n=19）为 2.6 kPa。我们还测试了细胞核和核仁染色有机染料对细胞机械性能的影响。对于染色的 EPC2 细胞 (n=5)，发现显着变硬 (9.9 kPa)，而 CP-A 细胞 (n=5) 无明显趋势 (2.9 kPa)，而 CP-D 细胞 (n=16) 则观察到轻微软化 (2.1 kPa)。一些力-压痕曲线显示出具有负斜率段的非单调不连续性，类似于锯齿图案。我们发现这些“突破事件”的发生率在发育异常的 CP-D 细胞中最高，在化生 CP-A 细胞中居中，在正常 EPC2 细胞中最低。这一观察结果表明，癌细胞和癌前细胞顺应性增加的微观解释可能在于它们对“崩溃和屈服”的敏感性，而不是它们“弯曲和弯曲”的能力。
The mechanical stiffness of individual cells is important in tissue homeostasis, cell growth, division, and motility, and the epithelial-mesenchymal transition in the initiation of cancer. In this work, a normal squamous cell line (EPC2) and metaplastic (CP-A) as well as dysplastic (CP-D) Barrett’s Esophagus columnar cell lines are studied as a model of pre-neoplastic progression in the human esophagus. We used the combination of an atomic force microscope (AFM) with a scanning confocal fluorescence lifetime imaging microscope (FLIM) to study the mechanical properties of single adherent cells. 64 force indentation curves were taken over the nucleus of each cell in an 8×8 grid pattern. Analyzing the force indentation curves, indentation depth dependent Young’s moduli were found for all cell lines. Stiffness tomograms demonstrate distinct differences between the mechanical properties of the studied cell lines. Comparing the stiffness for indentation forces of 1 nN, most probable Young’s moduli were calculated to 4.7 kPa for EPC2 (n=18 cells), 3.1 kPa for CP-A (n=10), and 2.6 kPa for CP-D (n=19). We also tested the influence of nuclei and nucleoli staining organic dyes on the mechanical properties of the cells. For stained EPC2 cells (n=5), significant stiffening was found (9.9 kPa), while CP-A cells (n=5) showed no clear trend (2.9 kPa) and a slight softening was observed (2.1 kPa) in the case of CP-D cells (n=16). Some force-indentation curves show non-monotonic discontinuities with segments of negative slope, resembling a sawtooth pattern. We found the incidence of these ‘breakthrough events’ to be highest in the dysplastic CP-D cells, intermediate in the metaplastic CP-A cells, and lowest in the normal EPC2 cells. This observation suggests that the microscopic explanation for the increased compliance of cancerous and pre-cancerous cells may lie in their susceptibility to ‘crumble and yield’ rather than their ability to ‘bend and flex’.
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影响因子：
29.4
作者：
Palanca-Wessels, MC;Barrett, MT;Rabinovitch, PS
通讯作者：
Rabinovitch, PS
影响因子：
3.5
作者：
Cross, Sarah E.;Jin, Yu-Sheng;Gimzewski, James K.
通讯作者：
Gimzewski, James K.
影响因子：
11.2
作者：
Remmerbach, Torsten W.;Wottawah, Falk;Guck, Jochen
通讯作者：
Guck, Jochen
影响因子：
1.6
作者：
HUTTER, JL;BECHHOEFER, J
通讯作者：
BECHHOEFER, J
粘着斑蛋白在三维细胞运动中的独特作用。
影响因子：
21.3
作者：
Fraley, Stephanie I.;Feng, Yunfeng;Krishnamurthy, Ranjini;Kim, Dong-Hwee;Celedon, Alfredo;Longmore, Gregory D.;Wirtz, Denis
通讯作者：
Wirtz, Denis