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Confocal Microscopy Image Gallery
Owl Monkey Kidney Epithelial Cells (OMK Line)
The owl monkey (Aotus trivirgatus) is a nocturnal native of tropical South America that is known for its large, closely set eyes. The OMK epithelial cell line was established from kidney tissue excised from a member of the species.
Studies indicate that OMK cells are susceptible to infection with several different non-human primate viruses, such as herpesvirus saimiri, herpesvirus ateles, and herpesvirus aotus. The line is commonly used to propagate these viruses for research purposes. OMK cells have also been utilized for other purposes, including studies of both the simian immunodeficiency virus (SIV) and the human immunodeficiency virus (HIV).
OMK cells naturally restrict the growth of HIV-1, but researchers have found that this restriction can be manipulated through treatment with certain pharmacological reagents. Investigations with the OMK line indicate that the cells increase their permissiveness to infection with HIV-1 by 100-fold when they are exposed to cyclosporine A (CsA), an immunosuppressant. CsA competitively inhibits cyclophilin A (CypA), one of the proteins that is incorporated into newly budded particles of HIV. The interaction between CypA and HIV-1 is thought to contribute to the restriction of the virus in the New World monkey cells. Notably, cell lines established from Old World primates do not experience the same extent of change in permissiveness of HIV-1 when treated with CsA as do OMK cells.
In a double immunofluorescence experiment, the adherent monolayer culture of owl monkey kidney cells illustrated above was fixed, permeabilized, blocked with 10-percent normal goat serum, and then treated with a cocktail of mouse anti-vimentin (intermediate filaments) and rabbit anti-GPP130 (Golgi complex) primary antibodies followed by goat anti-mouse and anti-rabbit secondary antibodies (IgG) conjugated to Alexa Fluor 568 (pseudocolored blue) and Alexa Fluor 488, respectively. Cell nuclei were counterstained with the red-absorbing dye TO-PRO-3. Images were recorded with a 60x oil immersion objective using a zoom factor of 2.5 and sequential scanning with the 488-nanometer spectral line of an argon-ion laser, the 543-nanometer line from a green helium-neon laser, and the 633-nanometer line of a red helium-neon laser. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles unless otherwise noted above.
Additional Confocal Images of Owl Monkey Kidney Epithelial (OMK) Cells
Triple Fluorophore Labeling of OMK Epithelial Cells with BODIPY FL, MitoTracker Orange CMTMRos, and TO-PRO-3 - The distribution of filamentous actin and mitochondria in an adherent culture of OMK epithelial cells was visualized with BODIPY FL conjugated to phallacidin and MitoTracker Orange CMTMRos (pseudocolored yellow). Cell nuclei were counterstained with TO-PRO-3, a carbocyanine monomer with long-wavelength red fluorescence.
Proximity Between the Nucleus and the Golgi Complex in Owl Monkey Kidney Cells - GPP130 is a transmembrane protein localized to the cis-Golgi. The culture of owl monkey kidney epithelial cells presented in this section was immunofluorescently labeled with anti-GPP130 rabbit monoclonal primary antibodies followed by goat anti-rabbit Fab fragments conjugated to Alexa Fluor 488. In addition, the specimen was stained for DNA in the nucleus with the red-absorbing dye TO-PRO-3.
Distribution of F-Actin and Mitochondria in OMK Epithelial Cells - The log phase culture of OMK epithelial cells presented in this section was labeled for F-actin and the intracellular mitochondrial network with Alexa Fluor 488 conjugated to phalloidin and MitoTracker Red CMXRos, respectively. The high-affinity DNA probe DRAQ5 (pseudocolored cyan) was utilized to counterstain cell nuclei.
Targeting the Golgi and Intermediate Filament Networks in OMK Cell Cultures with Immunofluorescence - A semi-confluent culture of owl monkey kidney cells was fixed, permeabilized, and blocked with 10-percent normal goat serum in phosphate-buffered saline prior to immunofluorescent labeling with rabbit primary antibodies to GPP130, a protein resident in the Golgi complex of mammalian cells, and mouse primary antibodies to vimentin, an intermediate filament protein. The culture was subsequently stained with a mixture of anti-rabbit and anti-mouse secondary antibodies conjugated to Alexa Fluor 568 (pseudocolored yellow) and Alexa Fluor 488, respectively.
Contributing Authors
Nathan S. Claxton, Shannon H. Neaves, and Michael W. Davidson - National High Magnetic Field Laboratory, 1800 East Paul Dirac Dr., The Florida State University, Tallahassee, Florida, 32310.