Pleomorphism in Biological Units of Life: Morphological Heterogeneity in Cells Does Not Translate Uniformly to Subcellular Components

The interplay of the three-dimensional (3D) distribution
of various
subcellular components and their interactions are expected to control
overall cellular morphology in biology. In this study, we aimed to
determine whether the pleomorphy observed at the whole-cell level
is being reflected by the components constituting the cells by focusing
on the 3D distribution of pixel intensities at the single-cell level
of the whole (cell) and its parts (the seven subcellular components
of the cellsself-assemblies of smaller units). We rigorously
acquired and analyzed the image data of RAW264.7 cells at the single-cell
level. We report asymmetries in the spatial distribution of pixel
intensities at the whole-cell and subcellular component levels along
with the occurrence of alterations when pleomorphism is reduced by
synchronization of the cell cycle. From our repertoire of seven subcellular
components, we report ER, mitochondria, and tubulin to be independent
of whole-cell apico-basal heterogeneity of optical density while nuclear,
plasma membrane, lysosomal, and actin fluorescence distributions are
found to contribute to the apico-basal polarity of the whole cell.
While doing so, we have also developed an image analysis algorithm
utilizing 2D segmentation to analyze the single cells in 3D using
confocal microscopy, a technique that allows us to analyze cellular
states in their native hydrated state.