14 4
There are also several other hydrophobic fluorescent dyes available that may
be useful in the quantitative assessment of adipogenesis, such as Bodipy
493/503. There are reports that Bodipy 493/503 is more specific than Nile Red,
but further studies are needed to validate this statement (Gocze and Freeman
1994 ). A definite advantage of Bodipy 493/503 is that it has a more specific
fluorescent emission spectrum range. This allows for easier combination of cell
surface markers with the lipid-specific dyes to study the phenotypic characteris-
tics of adipocytes in more detail.
Fig. 7.2 Adipogenic lineage microscopy analysis. (a) Oil Red O-stained and 1 % Toluidine Blue-
counterstained non-induced culture. The red arrow indicates Oil Red O residue overlaying the
culture. The cells are fibroblast-like, small, slender and elongated as indicated by the blue arrow
and conform to describe ASC morphology. (b) Oil Red O-stained and 1 % Toluidine Blue-
counterstained adipogenic induced culture. Visible Oil Red O droplets confirm differentiation
into the adipogenic linage. (c) A magnification snapshot of (b). The red arrow indicates the pres-
ence Oil Red O residue overlaying the cell. Yellow circles demonstrate adipocytes (cytoplasm
completely filled with lipid droplets and also fusion of the droplets with associated lipid vacuole
formation). Green rectangle demonstrates a more mature preadipocyte with incomplete lipid
droplet formation. Blue triangle demonstrates a preadipocyte with an enlarged triangular shaped
cell that conforms to the morphology of adipocytes but does not contain any visual Oil Red
O-stained lipid vacuoles
F.A. van Vollenstee et al.