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Confocal Microscopy Image Gallery
Human Brain Glioma Cells (U-118 MG Line)
From 1966 to 1969, multiple cell lines were established by J. Ponten and associates from human gliomas, including the U-118 MG line. The U-118 MG line, which has been found to be tumorigenic in nude mice inoculated subcutaneously, contains both glioblastoma and astrocytoma cells.
In the mid-1980s, it was discovered that the U-118 MG cell line was contaminated with mycoplasmata, but stocks of the cells were subsequently treated with BM-cycline for six weeks in order to eliminate the problem. Many characteristics of U-118 MG cells are analogous to those of U-138 MG glioblastoma cells, although the two lines were purportedly derived from different sources. Studies indicate that the cells share identical VNTR (variable number of tandem repeats) patterns, similar STR (short tandem repeat) patterns, and at least six derivative marker chromosomes.
Gliomas are cancerous growths that originate in glial tissue, which is generally considered to primarily play a supportive role to the neurons present in the nervous system. The tumors can form not only in the brain, but also in the heart, optic nerve, spinal cord, and many other locations in the body. The presenting symptoms of gliomas vary based on their location and rate of growth. When located in the brain, some of the most common symptoms include headaches, speech problems, seizures, weakness or paralysis on one side of the face or body, blurred vision, and changes in behavior or thought processes. Approximately 20,000 Americans are diagnosed with gliomas annually.
A triple fluorophore combination of MitoTracker Orange CMTMRos, BODIPY FL conjugated to phallacidin, and TO-PRO-3 was used to label an adherent culture of human brain glioma 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 subsequently labeled with the conjugated phallotoxin and counterstained with the cyanine monomer (TO-PRO-3) reagent. 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, the MitoTracker Orange CMTMRos image channel was pseudocolored yellow, while the BODIPY FL image channel was pseudocolored green and the TO-PRO-3 channel was pseudocolored red.
Additional Confocal Images of Human Brain Glioma (U-118 MG) Cells
U-118 MG Cells Triple Labeled with Alexa Fluor 488, MitoTracker Deep Red 633, and DRAQ5 - The single human glioma cell (U-118 MG) presented in this section was resident in an adherent culture stained for F-actin with Alexa Fluor 488 conjugated to phalloidin, and for the mitochondrial network with MitoTracker Red CMXRos. The culture was also treated with DRAQ5, a far-red fluorescent DNA probe.
Targeting F-Actin and the Mitochondrial Network in Human Brain Glioma Cell Cultures - The distribution of filamentous actin and mitochondria in a culture of U118-MG glioma 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.
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.