Fluorescence Digital Image Gallery

Normal African Green Monkey Kidney Epithelial Cells (Vero)

The production of antibodies in vitro was introduced by Georges K÷hler and CÚsar Milstein, who first employed cultures of fused cells to isolate antibodies of defined specificity. In this procedure, termed monoclonal antibody production, antibodies are first produced in mice, and activated B-lymphocytes from the host's spleen (the source of antibodies) are fused with cultured plasma tumor myeloma cells (that do not produce antibodies) from mice of the same strain. The fusion product is a hybrid (termed a hybridoma) of the two cells that continues to grow and divide in culture and produces large amounts of the antibody. Unfused cells and non-productive fusions are eliminated by classical cloning techniques to purify the culture of antibody-producing cells. A hybrid cell line produces only a single type of antibody, which can be used as a selection marker in serial dilution cloning by monitoring the culture medium for secreted antibody at each stage. Ultimately a cell line is established from a single cell (monoclonal) in which all of the cells produce an identical antibody.

Elements of the cytoskeletal and microtubule networks were imaged in the Vero cell culture illustrated above by treating the fixed and permeabilized cells with mouse anti-alpha-tubulin primary antibodies, followed by goat anti-mouse secondary antibodies conjugated to Alexa Fluor 568. Mixed with the secondary antibody cocktail was Alexa Fluor 350 conjugated to phalloidin to target filamentous actin. Nuclei were counterstained with SYTOX Green. 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 larger image of the African green monkey kidney epithelial (Vero) cells.

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