Fluorescence Digital Image Gallery

Embryonic Rat Thoracic Aorta Medial Layer Myoblast Cells (A-10)

The cytoskeletal actin filamentous network is anchored, in part, by focal adhesion contacts and adherens junctions, which are membrane-associated complexes that serve as nucleation sites for the filament termini. These centers also act as crosslinkers between the exterior cell matrix, plasma membrane, and actin cytoskeleton. Serving to connect the extracellular matrix to the actin cytoskeleton, the focal adhesion sites function as strategic structural elements. In addition, these complexes perform as sites of cellular signal transduction by initiating signaling pathways in response to new adhesion contacts. Among other proteins, focal adhesion junctions contain integrin-type receptors that are attached to the extracellular matrix and are associated with complexes containing vinculin (a universal focal adhesion marker), talin, alpha-actinin, tensin, paxillin, zyxin, and focal adhesion kinase.

Focal adhesions were visualized in the log phase adherent monolayer culture of A-10 cells illustrated above by immunofluorescent treatment with mouse anti-vinculin primary antibodies followed by goat anti-mouse Fab fragments conjugated to the cyanine dye, Cy2. The actin cytoskeletal network was simultaneously imaged with Alexa Fluor 568 conjugated to phalloidin, and nuclei were counterstained with Hoechst 33258. Images were recorded in grayscale with a QImaging Retiga Fast-EXi camera system coupled to an Olympus BX-51 microscope equipped with bandpass emission fluorescence filter optical blocks provided by Omega Optical. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles.

View a smaller image of the rat thoracic aorta (A-10) cell.

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