Determination of three-dimensional morphometry of adherent cells by surface profilometry

A three-dimensional, non-contacting surface profiler was used to analyse cells that had been previously prepared for scanning electron microscopy. The height of ten attached and spread cells was measured in both two- and three-dimensions (2-D and 3-D). The 2-D scan is a typical, linear profilometer output, while a 3-D image is produced with a 3-D detector. The 3-D contour plots of each cell were then compared with scanning electron microscopy micrographs by digitizing four morphological parameters: perimeter, surface area, long axis and short axis of each cell. A paired Student's t test showed that the two imaging techniques are not equivalent. Non-equivalence was attributed to a difference in the method of calibration. However, when combined with scanning electron microscopy, the profiler allowed complete three-dimensional quantitative analysis of cellular morphology.