Bone-marrow-derived mast cells were allowed to spread on a polylysine-coated coverslip. The cells were fixed, labeled with CellTrace™ CFSE dye (Invitrogen), and imaged at 40µm in PBS buffer. Imaged by Z. Deng, T. Zink, H. Chen, F. Liu, and Gang-yu Liu at UC Davis, and assisted by L. Yu and A. Hicklin, using the Asylum Research MFP-3D-CF™ AFM integrated with an Olympus FluoView™ 1000 confocal microscope.
This 3D image of mast cells combines two distinct yet correlated datasets. The AFM provides high-resolution surface information; the confocal shows the distribution of the fluorescent label throughout the cell. Note the four dark regions near the center left of the image; the upper two are beneath the cell surface, whereas the lower two are surface features visible in the topography.
A flythrough video shows an animation created in the IGOR Pro software:
Below are the separate datasets: confocal (left) and AFM topography (right). Data fusion was accomplished using ARgyle™, the advanced 3D rendering engine incorporated in the MFP-3D system software.