Human Bone Osteosarcoma Cells (U-2 OS Line)

The mitochondrion-selective stain MitoTracker Red CMXRos was utilized to label the mitochondrial network in a culture of U-2 OS cells (illustrated above), and the popular cell-permeant nuclear counterstain Hoechst 33342 was used to label cell nuclei. In addition, filamentous actin was targeted with Alexa Fluor 488 conjugated to phalloidin. Images were recorded in grayscale with a 12-bit digital camera coupled to either a Nikon E-600 or Eclipse 80i microscope equipped with bandpass emission fluorescence filter optical blocks. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles.