Fast, multi-frequency, and quantitative nanomechanical mapping of live cells using the atomic force microscope.
Fast, multi-frequency, and quantitative nanomechanical mapping of live cells using the atomic force microscope.
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DOI：
10.1038/srep11692
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发表时间：
2015-06-29
影响因子：
4.6
通讯作者：
Raman A
中科院分区：
文献类型：
--
作者：
Cartagena-Rivera AX;Wang WH;Geahlen RL;Raman A
细胞机械生物学的一个长期目标是将支撑疾病或形态发生的动态生物分子过程与纳米尺度力学性质的时空变化联系起来，如粘弹性、表面张力和粘附性。这就要求在单个细胞内发展具有高时空分辨率的定量机械显微镜方法。原子力显微镜(AFM)可以高空间分辨率地描绘细胞的异质力学性质，但图像采集时间比研究动态细胞过程所需的时间长1-2个数量级。我们提出了一种技术，使商用原子力显微镜系统能够在大范围的宽间隔频率(几个10‘S微米)，空间分辨率等于幅度调制(AM-AFM)，图像采集时间(几十秒)接近散斑荧光方法的情况下，定量地绘制活的真核细胞动态变化的粘弹性特性图。与AM-AFM相比，这意味着纳米机械成像吞吐量提高了约20倍，并且与新兴的高速AFM系统完全兼容。该方法用于研究乳腺癌细胞MDA-MB-231对Syk蛋白酪氨酸激酶抑制的时空力学反应，从而深入了解Syk负性调节高侵袭性癌细胞运动的信号通路。
A longstanding goal in cellular mechanobiology has been to link dynamic biomolecular processes underpinning disease or morphogenesis to spatio-temporal changes in nanoscale mechanical properties such as viscoelasticity, surface tension, and adhesion. This requires the development of quantitative mechanical microscopy methods with high spatio-temporal resolution within a single cell. The Atomic Force Microscope (AFM) can map the heterogeneous mechanical properties of cells with high spatial resolution, however, the image acquisition time is 1–2 orders of magnitude longer than that required to study dynamic cellular processes. We present a technique that allows commercial AFM systems to map quantitatively the dynamically changing viscoelastic properties of live eukaryotic cells at widely separated frequencies over large areas (several 10’s of microns) with spatial resolution equal to amplitude-modulation (AM-AFM) and with image acquisition times (tens of seconds) approaching those of speckle fluorescence methods. This represents a ~20 fold improvement in nanomechanical imaging throughput compared to AM-AFM and is fully compatible with emerging high speed AFM systems. This method is used to study the spatio-temporal mechanical response of MDA-MB-231 breast carcinoma cells to the inhibition of Syk protein tyrosine kinase giving insight into the signaling pathways by which Syk negatively regulates motility of highly invasive cancer cells.
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DOI：
10.1073/pnas.211400898
发表时间：
2001-10-23
影响因子：
11.1
作者：
Ando, T;Kodera, N;Toda, A
通讯作者：
Toda, A
影响因子：
4.8
作者：
Gallet, C;Rosa, JP;Maclouf, J
通讯作者：
Maclouf, J
影响因子：
3.4
作者：
Alcaraz, J;Buscemi, L;Navajas, D
通讯作者：
Navajas, D
影响因子：
3
作者：
Heu, Celine;Berquand, Alexandre;Nicod, Laurence
通讯作者：
Nicod, Laurence
影响因子：
4
作者：
Humphris, ADL;Miles, MJ;Hobbs, JK
通讯作者：
Hobbs, JK