Navigation Links
Carnegie Mellon scientists create rainbow of fluorescent probes
Date:3/25/2010

SAN FRANCISCOScientists at Carnegie Mellon University's Department of Chemistry and Molecular Biosensor and Imaging Center (MBIC) are advancing the state-of-the-art in live cell fluorescent imaging by developing a new class of fluorescent probes that span the spectrum from violet to the near-infrared. The new technology, called fluoromodules, can be used to monitor biological activities of individual proteins in living cells in real time. At the 239th national meeting of the American Chemical Society, Carnegie Mellon chemists and MBIC scientists will discuss recent advances in their fluoromodule technology that have produced diverse and photostable probes.

Fluoromodules, which consist of dye-protein complexes, provide alternatives to common fluorescent proteins, such as Green Fluorescent Protein (GFP), but with a wider selection of colors and the potential for significantly greater photostability, which allows scientists to image the dye for longer periods of time. This is made possible by the fact that the dye is noncovalently bound to the protein, which allows fresh dye to replace bleached dye.

"We initially isolated and characterized fluoromodules that generate fluorescence from the fluorogenic dyes thiazole orange and malachite green. We are now expanding our repertoire by synthesizing new dyes that emit in the orange and violet regions of the spectrum, and engineering proteins that bind to the new dyes with great affinity," said Chemistry Professor Bruce Armitage, co-director of the Center for Nucleic Acid Science and Technology at Carnegie Mellon and a member of the MBIC team developing the fluoromodules.

Fluoromodules are made of a fluorogen-activating protein (FAP) and a non-fluorescent dye called a fluorogen. The FAP, which is genetically expressed in a cell and tagged to a protein of interest, does not become fluorescent until it binds with its fluorogen. With the novel FAPs and associated fluorogens created by the MBIC team, the researchers can control when a target protein lights up, allowing them to track proteins on the cell surface and within living cells in very simple and direct ways, eliminating cumbersome experimental steps.

Recent advances in the MBIC fluoromodule technology being presented at the ACS meeting include:

  • Working with a FAP that had a low affinity for the fluorogenic dye dimethlindole red (DIR), graduate student Hayriye zhalici-nal used PCR mutagenesis to introduce mutations into the FAP's genetic sequence. A small number of mutations increased several-fold the protein's affinity for DIR, enabling very specific and selective binding of the FAP to its dye partner (DIR). zhalici-nal will present this work at 9:50 a.m., Thursday, March 25 during the Follow-on Biologics: Protein Engineering session located in room 201 West Bldg. in the Moscone Center.

  • Graduate student Nathaniel Shank synthesized a modified DIR, making it eight-times more photostable. This significant improvement could have an impact on single molecule imaging. Additionally, the modified DIR emits in the orange range of the spectrum, adding another color to the fluoromodule toolkit being developed at MBIC. Shank will present this work at 8 p.m., Tuesday, March 23 during the Total Synthesis of Complex Molecules, Material Devices & Switches, Physical Organic Chemistry poster session located in Hall D of the Moscone Center.

  • By synthesizing a new dye and identifying FAPs that bind to it, research chemist Gloria Silva and graduate student Kim Zanotti developed a fluoromodule that emits fluorescence in the violet, which is a welcome addition to a very limited number of probes able to emit in the violet portion of the spectrum. Zanotti will present this work a 6 p.m., Tuesday, March 23 during the poster session located in room 3009/3011 West Bldg. in the Moscone Center.


'/>"/>

Contact: Jocelyn Duffy
jhduffy@andrew.cmu.edu
412-600-0029
Carnegie Mellon University
Source:Eurekalert

Related biology technology :

1. 2010 Carnegie Science Awardees Announced
2. Carnegie Mellon MRI technology that noninvasively locates, quantifies specific cells in the body
3. MIT scientists transform polyethylene into a heat-conducting material
4. Scientists discover how ocean bacterium turns carbon into fuel
5. Scientists glimpse nanobubbles on super nonstick surfaces
6. Princeton scientists find an equation for materials innovation
7. Scientists glimpse nanobubbles on super non-stick surfaces
8. Scientists transplant nose of mosquito, advance fight against malaria
9. Penn material scientists turn light into electrical current using a golden nanoscale system
10. Seeing the quantum in chemistry: JILA scientists control chemical reactions of ultracold molecules
11. NFCR Scientists Discover Brain Tumor's “Escape Path”
Post Your Comments:
*Name:
*Comment:
*Email:
(Date:4/29/2016)... ... 29, 2016 , ... Amendia, Inc., a leading designer, developer, ... the completion of a significant transaction and partnership that positions Amendia for accelerated ... Kohlberg & Company, L.L.C. (“Kohlberg”), a leading private equity firm specializing in ...
(Date:4/29/2016)... Louisville, KY and San Diego, CA (PRWEB) , ... April 29, ... ... the National Stem Cell Foundation (NSCF) to support the development of a patient-specific stem ... by Dr. Andrés Bratt-Leal in the lab of Dr. Jeanne Loring at The Scripps ...
(Date:4/28/2016)... PUNE, India , April 28, 2016 ... PT, JT, Stirling, and Brayton Cryocoolers), Service (Technical Support, ... Application, and Geography - Global Forecast to 2022", published ... to USD 2.94 Billion by 2022, at a CAGR ... Browse 70 market data Tables and 94 Figures spread ...
(Date:4/28/2016)... ... 2016 , ... Next week on May 5 at 2:55 ... for tissue stem cell counting and expansion to gene-editing scientists and other attendees ... Genome Engineering in Burlington, Massachusetts. , The attention of most gene-editing scientists appears ...
Breaking Biology Technology:
(Date:3/10/2016)... --  Unisys Corporation (NYSE: UIS ) today announced ... testing its biometric identity solution at the Otay Mesa border ... identify certain non-U.S. citizens leaving the country. ... determine the efficiency and accuracy of using biometric technologies in ... until May 2016. --> the United States ...
(Date:3/3/2016)... , March 3, 2016  FlexTech, a SEMI Strategic ... of Innovation, Research & Development, Leadership in Education, and, ... is the 9 th year of the FLEXI ... companies and individuals from past years . Judging ... a pre-described set of criteria, by a panel of ...
(Date:3/1/2016)... , March 1, 2016 /PRNewswire/ ... announced the addition of the  "Global ...  report to their offering. ... the addition of the  "Global Biometric ...  report to their offering. --> ...
Breaking Biology News(10 mins):