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.
The flythrough video shows an animation created in the IGOR Pro software as you zoom in and around the surface of the 3D image. Play the video:
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.
Back
