1993 Volume 1 Issue 1 Pages 1-8
In this paper we describe an application of the confocal laser microscope (CLM) to interference reflection (IR) microscopy for analysis of the cell-to-substrate adhesion of cultured cells. The CLM in IR mode provided sharp and high-contrast images of focal contacts of cells grown on glass surfaces without interference from out-of-focus reflections. For simultaneous imaging of fluorescence and IR signals, the 488 nm line of an argon ion laser was used for illumination and a dichroic mirror was placed in front of two photomultipliers to split the light returning from the specimen into reflected (488 nm) and fluorescence (green) signals. Thus we could correlate the IR and fluorescence images of cells stained with fluorescent labels for vinculin or F-actin. To confirm the cell contact, we also examined cultured living cells perfused with a medium containing Lucifer Yellow. Fluorescent dye-excluded spots in the plane of cell adhesion corresponded well to dark spots in IR images. Because of these advantages over the conventional IR microscope, we can corroborate the usefulness of the CLM for IR analysis.
