Fluorescence Digital Image Gallery

Human Bone Osteosarcoma Cells (U-2 OS)

Mutagenesis experiments with green fluorescent protein have produced a large number of variants with improved folding and expression characteristics, which have eliminated wild-type dimerization artifacts and fine-tuned the absorption and fluorescence properties. One of the earliest variants, known as enhanced green fluorescence protein (EGFP), contains codon substitutions (commonly referred to as the S65T mutation) that alleviates the temperature sensitivity and increases the efficiency of GFP expression in mammalian cells. Proteins fused with EGFP can be observed at low light intensities for long time periods with minimal photobleaching. Enhanced green fluorescent protein fusion products are optimally excited by the 488-nanometer spectral line from argon and krypton-argon ion lasers in confocal microscopy. This provides an excellent biological probe and instrument combination for examining intracellular protein pathways along with the structural dynamics of organelles and the cytoskeleton.

In order to generate the digital image featured above, stable U-2 OS transfectants containing a subcellular mitochondrial localization vector of DsRed fluorescent protein fused to the mitochondrial targeting sequence of human cytochrome C oxidase were fixed, permeabilized, and blocked with bovine serum albumen before being labeled with Alexa Fluor 350 conjugated to phalloidin. The nuclei were counterstained with SYTOX Green. Images were recorded in grayscale with a QImaging Retiga Fast-EXi camera system coupled to an Olympus BX-51 microscope equipped with bandpass emission fluorescence filter optical blocks provided by Omega Optical. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles.

View a larger image of the human bone osteosarcoma (U-2 OS) cells.

© 1995-2022 by Michael W. Davidson and The Florida State University. All Rights Reserved. No images, graphics, software, scripts, or applets may be reproduced or used in any manner without permission from the copyright holders. Use of this website means you agree to all of the Legal Terms and Conditions set forth by the owners.
This website is maintained by our
Graphics & Web Programming Team
in collaboration with Optical Microscopy at the
National High Magnetic Field Laboratory.
Last modification: Thursday, Oct 14, 2004 at 08:34 AM
Access Count Since July 16, 2004: 10340
Microscopes, fluorescence filters, and digital imaging equipment provided by:
Visit the Olympus Microscopy Resource Center website. Visit the Omega Optical website. Visit the QImaging website.