Barr Valarie A, Bunnell Stephen C
National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.
Curr Protoc Cell Biol. 2009 Dec;Chapter 4:Unit 4.23. doi: 10.1002/0471143030.cb0423s45.
Interference reflection microscopy (IRM) is an optical technique used to study cell adhesion or cell mobility on a glass coverslip. The interference of reflected light waves generates images with high contrast and definition. IRM can be used to examine almost any cell that will rest upon a glass surface, although it is most useful in examining sites of close contact between a cell and substratum. This unit presents methods for obtaining IRM images of cells with particular emphasis on IRM imaging with a laser scanning confocal microscope (LSCM), as most LSCM are already capable of recording these images without any modification of the instrument. Techniques are presented for imaging fixed and live cells, as well as simultaneous multi-channel capture of fluorescence and reflection images.
干涉反射显微镜(IRM)是一种光学技术,用于研究玻璃盖玻片上的细胞黏附或细胞运动。反射光波的干涉产生具有高对比度和清晰度的图像。IRM可用于检查几乎任何能在玻璃表面上静止的细胞,尽管它在检查细胞与基质紧密接触的部位时最为有用。本单元介绍了获取细胞IRM图像的方法,特别强调了使用激光扫描共聚焦显微镜(LSCM)进行IRM成像,因为大多数LSCM已经能够在不对仪器进行任何修改的情况下记录这些图像。文中介绍了固定细胞和活细胞成像技术,以及荧光和反射图像的同步多通道采集技术。