Abstract
We present a new technique for estimating cell surface geometry. A dish supporting adherent cells is observed using oblique transillumination and rotated in the horizontal plane using a stepping motor. The stage rotation-dependent movements of the start and end points of a shadow formed behind the illuminated cells uniquely determine the relative height differences between points along the cell surface. Thus, using custom-made apparatuses and living endothelial cells, we demonstrate that the combination of a rotating stage and oblique lighting allows for the evaluation of three-dimensional surface geometry of adherent cells. As compared to confocal microscopy and atomic force microscopy, which are commonly used for measuring cell surface geometry, this approach can be performed rapidly and is especially suitable for the observation of unstained cells over a large surface covering multiple cells at a time.
Highlights
Cell morphology and its function are closely associated [1]
Using custom-made apparatuses and living endothelial cells, we demonstrate that the combination of a rotating stage and oblique lighting allows for the evaluation of three-dimensional surface geometry of adherent cells
The shadowing effect from the resultant oblique lighting provided cell images similar in appearance to those taken by differential interference contrast (DIC) microscopy and reflected light oblique transillumination microscopy [6] (Figure 2)
Summary
Cell morphology and its function are closely associated [1]. observing the cell morphology is common in cellular biology. The height and slope in the z-direction (vertical direction) of cells adhering to a culture dish or plate via the extracellular matrix cannot be measured by ordinary optical and electron microscopy [2]. Confocal microscopy and atomic force microscopy are used to observe the three-dimensional morphology of adherent cells [3,4,5]. Confocal microscopy is an optical imaging technique used to increase optical resolution by using a spatial pinhole to eliminate out-offocus light in cells that are thicker than the focal plane. It enables the reconstruction of three-dimensional struc-. Cells were seeded in a 35mm diameter plastic base culture dish
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