Burmeister J S, Olivier L A, Reichert W M, Truskey G A
Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA.
Biomaterials. 1998 Mar;19(4-5):307-25. doi: 10.1016/s0142-9612(97)00109-9.
Cell adhesion and function depend upon the formation of adhesive contacts between the cell and substrate. Determination of the cell substrate contact area is necessary in order to understand how biomaterial properties influence cell adhesion. In this review we describe the development and application of total internal reflection fluorescence microscopy (TIRFM) to quantify the separation distance of cells from a biomaterial surface. An approximate theory is presented for the straightforward calculation of separation distances when a fluor is placed in the cell membrane. The validity of this approach is discussed. TIRFM is compared to interference reflection microscopy and related techniques that measure cell/substrate separation distances. This approach is then applied to a number of important problems in cell substrate interactions, including changes in contact area and adhesion strength on biomaterial surfaces, analysis of bond strength, and real-time measurement of cell/substrate separation distances following exposure to flow.
细胞黏附与功能取决于细胞与底物之间黏附接触的形成。为了理解生物材料特性如何影响细胞黏附，确定细胞与底物的接触面积是必要的。在本综述中，我们描述了全内反射荧光显微镜（TIRFM）的发展与应用，以量化细胞与生物材料表面的分离距离。当荧光团置于细胞膜中时，提出了一种用于直接计算分离距离的近似理论。讨论了该方法的有效性。将TIRFM与测量细胞/底物分离距离的干涉反射显微镜及相关技术进行了比较。然后将该方法应用于细胞与底物相互作用中的一些重要问题，包括生物材料表面接触面积和黏附强度的变化、键强度分析以及暴露于流动后细胞/底物分离距离的实时测量。
