通过延时共聚焦显微镜对在不同钛表面生长的活细胞进行分析。

Analysis of living cells grown on different titanium surfaces by time-lapse confocal microscopy.

作者信息

Gatti R, Orlandini G, Uggeri J, Belletti S, Galli C, Raspanti M, Scandroglio R, Guizzardi S

机构信息

Department of Experimental Medicine, University of Parma, Via Volturno 39, 43100 Parma, Italy.

出版信息

Micron. 2008;39(2):137-43. doi: 10.1016/j.micron.2006.11.009. Epub 2006 Dec 28.

DOI:10.1016/j.micron.2006.11.009

PMID:17223563

Abstract

In this study we have combined fluorescence- and reflection-confocal laser scanning microscopy for the simultaneous visualization of living cells and surface topography beneath them. To this purpose we have designed a specific flow chamber and we have tested it with osteoblasts grown on an opaque, thick support, made of smooth or sandblasted titanium. Cells were loaded with Calcein-AM or tetramethylrhodamine methyl ester (TMRM), two probes employed as indicators of cell viability/morphology and mitochondrial membrane potential, respectively. Besides the acquisition of stacks of confocal sections, the system allowed also vertical views and faithful three-dimensional reconstruction of the samples. Confocal microscope implemented with our flow chamber proved to be a promising tool for time-lapse investigation of cell-biomaterial interactions.

摘要

在本研究中，我们将荧光共聚焦激光扫描显微镜和反射共聚焦激光扫描显微镜相结合，以便同时观察活细胞及其下方的表面形貌。为此，我们设计了一种特殊的流动腔，并使用生长在由光滑或喷砂处理的钛制成的不透明厚支撑物上的成骨细胞对其进行了测试。细胞分别用钙黄绿素 - AM或四甲基罗丹明甲酯（TMRM）进行加载，这两种探针分别用作细胞活力/形态和线粒体膜电位的指示剂。除了采集共聚焦切片堆栈外，该系统还允许对样品进行垂直观察和逼真的三维重建。事实证明，配备我们所设计流动腔的共聚焦显微镜是用于细胞 - 生物材料相互作用延时研究的一种很有前景的工具。