Verschueren H
J Cell Sci. 1985 Apr;75:279-301. doi: 10.1242/jcs.75.1.279.
Since its introduction into cell biology by Curtis in 1964, interference reflection microscopy (IRM) has been used by an increasing number of researchers to study cell-substrate interactions in living cells in culture. With the use of antiflex objectives, high-contrast IRM images can now be readily obtained. From the different theories on image formation in IRM that have been put forward, it can be seen that a zero-order interference pattern is generated at high illuminating numerical aperture. This yields information on the closeness of contact between cell and substrate, with only minor perturbation by reflections from the dorsal cell surface. Therefore, the proper use of illuminating apertures is crucial. Nevertheless, IRM images have to be interpreted with caution, especially under thin cytoplasmic sheets. Quantitative IRM is possible only with a mathematical model for finite illuminating aperture interferometry and with an independent measurement of cell thickness for values up to 1 micron. IRM has been applied qualitatively to a large number of cell types, and it seems that there are two universal types of adhesion. Focal contacts are small regions of closest cell-substrate apposition, possibly of immediate contact, that are associated with the distal end of actin filament bundles. They are firm attachment structures that hold the cell in place and in its spread shape. Close contacts are broad areas of reduced cell-to-substrate distance. They are weaker but highly dynamic adhesions that sustain rapid movements of cells or cell parts over the substrate. Although a number of independent observations suggest that adhesion patterns of malignantly transformed cells differ from those of their normal counterparts, there is no simple correlation between malignancy in vivo and altered contact formation in vitro. The adhesion pattern seems to be determined by the locomotory state of the cells rather than by their tissue of origin. Finally, IRM can also be used to enhance contrast in images of fixed preparations.
自1964年柯蒂斯将干涉反射显微镜（IRM）引入细胞生物学以来，越来越多的研究人员使用它来研究培养中的活细胞与底物的相互作用。使用抗挠曲物镜，现在可以很容易地获得高对比度的IRM图像。从已提出的关于IRM图像形成的不同理论可以看出，在高照明数值孔径下会产生零级干涉图样。这产生了关于细胞与底物之间接触紧密程度的信息，而细胞背表面的反射造成的干扰很小。因此，正确使用照明孔径至关重要。然而，对IRM图像的解释必须谨慎，特别是在薄细胞质片层下。只有使用有限照明孔径干涉测量的数学模型并独立测量细胞厚度（值可达1微米），才可能进行定量IRM。IRM已被定性地应用于大量细胞类型，似乎存在两种普遍的黏附类型。黏着斑是细胞与底物最紧密并置的小区域，可能是直接接触区域，与肌动蛋白丝束的远端相关。它们是将细胞固定在适当位置并保持其伸展形状的牢固附着结构。紧密接触是细胞与底物距离减小的广阔区域。它们是较弱但高度动态的黏附，维持细胞或细胞部分在底物上的快速移动。尽管许多独立观察表明恶性转化细胞的黏附模式与其正常对应物不同，但体内恶性程度与体外接触形成改变之间没有简单的相关性。黏附模式似乎由细胞的运动状态而非其起源组织决定。最后，IRM还可用于增强固定制剂图像的对比度。
