Fluorescence Digital Image Gallery

Mongoose Skin Fibroblast Cells (APM)

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.

A fixed and permeabilized culture of adherent APM cells grown on cover glasses (illustrated above) was treated with mouse anti-vimentin monoclonal primary antibodies followed by goat anti-mouse secondary antibodies (IgG) conjugated to Texas Red-X. The filamentous actin network and DNA in the nuclei were simultaneously imaged by labeling with Oregon Green 488 conjugated to phalloidin and Hoechst 33258, respectively. 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 mongoose skin fibroblast (APM) cells.

© 1995-2019 by Michael W. Davidson and The Florida State University. All Rights Reserved. No images, graphics, software, scripts, or applets may be reproduced or used in any manner without permission from the copyright holders. Use of this website means you agree to all of the Legal Terms and Conditions set forth by the owners.
This website is maintained by our
Graphics & Web Programming Team
in collaboration with Optical Microscopy at the
National High Magnetic Field Laboratory.
Last modification: Thursday, Oct 14, 2004 at 08:34 AM
Access Count Since July 16, 2004: 6342
Microscopes, fluorescence filters, and digital imaging equipment provided by:
Visit the Olympus Microscopy Resource Center website. Visit the Omega Optical website. Visit the QImaging website.