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UC San Diego Scientists Chart Rapid Advances of Fluorescent Tools for Life-Science Research

Techniques Promise to Illuminate Details of Protein Expression, Activity and Function

Science Cover
Cells in the limelight on the cover of Science :
Parallel application of new (genetic tagging)
and old targeting methods and fluorophores.
Live cells were transfected with genetically
encoded fluorescent protein fused to tubulin in
order to lluminate the cells' microtubule networks
(green) while tetracysteine-actin and ReAsH
report on the location of stress fibers (red). Cells
were subsequently preserved and immuno-
labeled for the Golgi-matrix protein giantin using
quantum dots (yellow). DNA was subsequently
stained with Hoechst (blue). [Image: NCMIR/ Ben
Giepmans] Courtesy: AAAS/Science
San Diego, CA, April 14, 2006 -- An interdisciplinary team of biological imaging experts from the University of California , San Diego has published a review of fluorescent imaging technologies and underscored the importance of those technologies to major advances in the life sciences.

The article -- "The  Fluorescent Toolbox for Assessing Protein Location and Function" -- is the cover story in the April 14 issue of the journal Science .

"Fluorescent imaging is critical to the observation of dynamic processes in living systems," said lead author Ben Giepmans, a research scientist in the UCSD-based National Center for Microscopy and Imaging Research (NCMIR). "Some of these techniques now also allow researchers to localize the responsible molecular machine in situ by electron microscopy." Giepmans' co-authors on the Science paper include Roger Tsien, professor of pharmacology, chemistry and biochemistry; pharmacology project scientist Stephen Adams; and NCMIR director and UCSD School of Medicine neurosciences professor Mark Ellisman. Ellisman is a long-time participant in Calit2 and co-PI on the Calit2-led OptIPuter project. He is also an adjunct professor of bioengineering in the Jacobs School of Engineering, and directs the Center for Research in Biological Systems (CRBS). 

The National Institutes of Health and the Howard Hughes Medical Institute supported work directly related to this review. In their survey, the scientists contrasted the characteristic benefits and limitations of many new classes of fluorescent probes for studying proteins, including quantum dots, fluorescent proteins, and some genetic tags. Color-rich photomicrographs now routinely appear in scientific journals to illustrate dynamic biochemical processes. Those processes range from the expression of a specific gene to the redistribution of protein within a living cell.

Smarr and Ellisman
National Center for Microscopy and Imaging
Research director Mark Ellisman (right) with
CSE professor Larry Smarr in front of a
NCMIR image on Calit2's 105-megapixel display.

Progress in developing new fluorescent probes over the last decade has been dramatic. "Whole new classes of fluorescent dyes, fluorescent proteins, and other hybrid probes are being engineered to illuminate specific biochemical structures and processes within living cells," said Ellisman. "They also make possible the direct correlated imaging of the underlying molecular complexes at higher resolution by electron microscopy." 
 
Fluorescence imaging is rapidly becoming a biochemist's tool of choice for studying processes within living cells. Its rapid expansion is partially tied to a synergy of developments, including the increasing ease of implementing innovative targeting strategies to key cell metabolites and structures.  Concomitant advances in instrumentation and data analysis are enabling scientists to identify and quantify dynamic biochemical processes of living cells under light and electron microscopes. Fluorescence techniques are being adapted for clinical and biochemical assays like biopsies and high-throughput drug screening, and are just beginning to find wider application in functional assays of living cells and animals.

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