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Confocal Microscopy Image Gallery
Rabbit Kidney Epithelial Cells (RK13 Line)
The RK13 cell line is often employed in transfection experiments and to isolate viruses. The epithelial line was established from the renal tissue of a 5-week-old rabbit (Oryctolagus cuniculus).
RK13 cells are susceptible to infection with a wide variety of viruses, including herpes simplex, pseudorabies, vaccinia, rabbitpox, myxoma, simian adenoviruses, the B virus, and rubellavirus. Testing in the mid-1990s revealed that the cell line is positive for the bovine viral diarrhea virus (BVDV). The cells also are positive for the protein keratin.
The kidneys are organs specialized for filtration of the blood. Similar to the filtration system of a car, if the kidneys become clogged or overworked, a multitude of problems may occur. For instance, renal failure, diabetes, liver malfunction, and many other disorders may occur if toxins are allowed to build up in the blood because the kidneys are not able to work properly. In rabbits, such kidney-related ailments are not uncommon, the many toxins present in the environment, most of which have been introduced by humans, causing serious and widespread health problems for the small animals. Kidney problems in rabbits may also occur due to other causes, such as parasitic infection. Notably, the rabbit parasite Encephalitozoon cuniculi is only able to replicate in the kidneys and the host is only contagious when it is present in the organs. In the kidneys, E. cuniculi typically causes scarring and pitting, but when the parasite is passed through the blood to other areas of the body even more serious damage may occur, particularly paralysis related to infection of neural tissues.
The rabbit kidney cell culture presented above was immunofluorescently labeled with mouse anti-alpha-tubulin primary antibodies followed by goat anti-mouse secondary antibodies conjugated to Alexa Fluor 488. In addition, the specimen was stained with Alexa Fluor 546 conjugated to phalloidin (pseudocolored blue) and TO-PRO-3 (pseudocolored red), targeting the filamentous actin network and cell nuclei, respectively. Images were recorded with a 60x oil immersion objective using a zoom factor of 1.5 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 Rabbit Kidney Epithelial (RK13) Cells
Visualizing the Proximity Between the Microtubule Network and the Nucleus in RK13 Cells with Immunofluorescence - The details of the microtubule network were visualized in a culture of RK13 epithelial cells by employing immunofluorescence. The culture was labeled with mouse anti-alpha-tubulin followed by a secondary antibody (goat anti-mouse IgG) conjugated to Alexa Fluor 488. TO-PRO-3 (pseudocolored red) was utilized as a nuclear counterstain.
DsRed Fluorescent Protein in Transfected Rabbit Kidney Epithelial Cell Cultures - After transiently transfecting an adherent culture of RK13 rabbit kidney cells (DsRed2-Endoplasmic Reticulum; 10-percent transfection efficiency), the culture was washed, fixed, permeabilized, and blocked with bovine serum albumen. The cells were subsequently labeled with Alexa Fluor 488 conjugated to phalloidin and counterstained with Hoechst 33342 (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.