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Confocal Microscopy Image Gallery
Cat Kidney Cortex Epithelial Cells (CRFK Line)
The CRFK feline kidney cell line was established by a team led by R. A. Crandell from the renal cortex of a 12-week-old female cat (Felis catus domesticus). The cells, which are often utilized in viral research and in the production of vaccines, exhibit typical epithelial morphology.
Testing indicates that the CRFK line is free from the bovine viral diarrhea (BVD) virus that was previously found to have contaminated major stocks of a large number of permanent cell lines. The cells are susceptible, however, to the BVD virus, as well as an array of other viruses, including feline herpesvirus, feline calicivirus, feline panleukopenia, feline reovirus, canine parvovirus, and rabies virus.
The health of cats is of significant concern to much of the human population, who consider the animals cherished pets and companions. Thus, research focusing upon feline diseases and medical conditions flourishes and cat vaccinations are common. Many of the vaccines administered to cats are commonly developed in cultures of CRFK cells. Recently some concern has arisen regarding this practice, scientific evidence suggesting that CRFK-grown vaccines may be involved in the relatively high incidence of chronic renal failure in cats. Though further studies regarding the potential relationship between feline vaccines and diseases are needed, it has been demonstrated that administration of certain vaccines to cats can induce antibody responses to CRFK proteins and feline renal tissues.
A triple fluorophore combination of MitoTracker Orange CMTMRos (pseudocolored yellow), BODIPY FL conjugated to phallacidin, and DRAQ5 was used to label an adherent log phase culture of CRFK cells for mitochondria, the F-actin network, and nuclear DNA, respectively. The cells were first treated with the MitoTracker probe in growth medium for one hour, washed and fixed with paraformaldehyde (prepared in growth medium), permeabilized, and blocked with bovine serum albumen. The cells were then labeled with the conjugated phallacidin and counterstained with the anthraquinone (DRAQ5) reagent. Images were recorded with a 60x oil immersion objective using a zoom factor of 2.0 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 Cat Kidney Cortex Epithelial Cells (CRFK) Cells
Immunofluorescently Targeting the Golgi Complex in Feline Kidney Cell Cultures - The culture of feline kidney (CRFK) cells shown in this confocal image was fixed with paraformaldehyde, permeabilized, and treated with rabbit (anti-giantin) primary antibodies, followed by secondary antibodies conjugated to Alexa Fluor 568, targeting the Golgi apparatus. DNA in the cell nucleus was counterstained with TO-PRO-3 (pseudocolored cyan).
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.