Expansion Microscopy for Imaging the Cell-Material Interface.
Expansion Microscopy for Imaging the Cell-Material Interface.
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用于成像细胞材料界面的膨胀显微镜。
DOI：
10.1021/acsnano.1c11015
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发表时间：
2022-05-24
期刊：
影响因子：
17.1
通讯作者：
Cui B
中科院分区：
文献类型：
--
作者：
Nakamoto ML;Forró C;Zhang W;Tsai CT;Cui B
几十纳米到几微米尺度的表面形貌显著影响细胞粘附、迁移和分化。最近使用电子显微镜和超分辨率显微镜的研究提供了细胞如何与表面纳米形貌相互作用的见解;然而，这些方法所需的复杂样品制备和昂贵的成像设备使它们不容易获得。膨胀显微镜（ExM）是一种负担得起的方法，图像超过衍射极限，但ExM不能很容易地应用于图像的细胞-材料界面，因为大多数材料不膨胀。在这里，我们开发了一个协议，允许使用ExM解决细胞材料界面的高分辨率。我们应用该技术对U2 OS细胞和纳米结构基底之间的界面以及原代成骨细胞与钛牙种植体之间的界面进行成像。ExM的高空间分辨率显示，尽管AP 2和F-肌动蛋白都在垂直纳米结构诱导的弯曲膜上积累，但它们在空间上是分离的。使用ExM，我们还可靠地成像成骨细胞如何与低于衍射极限的粗糙钛种植体表面相互作用;这对于了解种植体的骨整合非常有意义，但到目前为止，由于钛种植体的不规则形状，大体积和不透明性，使其与其他超分辨率技术不兼容，因此一直是一个重大的技术挑战。我们相信，我们的协议将使使用ExM作为一个强大的工具，细胞材料界面的研究。
Surface topography on the scale of tens of nanometers to several micrometers substantially affects cell adhesion, migration, and differentiation. Recent studies using electron microscopy and super-resolution microscopy provide insight into how cells interact with surface nanotopography; however, the complex sample preparation and expensive imaging equipment required for these methods makes them not easily accessible. Expansion microscopy (ExM) is an affordable approach to image beyond the diffraction limit, but ExM cannot be readily applied to image the cell–material interface as most materials do not expand. Here, we develop a protocol that allows the use of ExM to resolve the cell–material interface with high resolution. We apply the technique to image the interface between U2OS cells and nanostructured substrates as well as the interface between primary osteoblasts with titanium dental implants. The high spatial resolution enabled by ExM reveals that although AP2 and F-actin both accumulate at curved membranes induced by vertical nanostructures, they are spatially segregated. Using ExM, we also reliably image how osteoblasts interact with roughened titanium implant surfaces below the diffraction limit; this is of great interest to understand osseointegration of the implants but has up to now been a significant technical challenge due to the irregular shape, the large volume, and the opacity of the titanium implants that have rendered them incompatible with other super-resolution techniques. We believe that our protocol will enable the use of ExM as a powerful tool for cell–material interface studies.
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光学成像。膨胀显微镜。
DOI：
10.1126/science.1260088
发表时间：
2015-01-30
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Chen F;Tillberg PW;Boyden ES
通讯作者：
Boyden ES
影响因子：
17.1
作者：
Chen W;Villa-Diaz LG;Sun Y;Weng S;Kim JK;Lam RH;Han L;Fan R;Krebsbach PH;Fu J
通讯作者：
Fu J
影响因子：
10.8
作者：
Li X;Klausen LH;Zhang W;Jahed Z;Tsai CT;Li TL;Cui B
通讯作者：
Cui B
影响因子：
46.9
作者：
Ku, Taeyun;Swaney, Justin;Park, Jeong-Yoon;Albanese, Alexandre;Murray, Evan;Cho, Jae Hun;Park, Young-Gyun;Mangena, Vamsi;Chen, Jiapei;Chung, Kwanghun
通讯作者：
Chung, Kwanghun
影响因子：
9.7
作者：
Le Guehennec, Laurent;Lopez-Heredia, Marco-Antonio;Layrolle, Pierre
通讯作者：
Layrolle, Pierre