Carnegie Mellon University scientists have married bright fluorescent dye molecules with DNA nanostructure templates to make nanosized fluorescent labels that hold considerable promise for studying fundamental chemical and biochemical reactions in single molecules or cells. The work, published online Jan. 26 in "The Journal of the American Chemical Society,” improves the sensitivity for fluorescence-based imaging and medical diagnostics.
"Our DNA nanotags offer unprecedented densities of fluorescent dyes and, thus, the potential for extremely bright fluorescent labels,” said graduate student Andrea Benvin, who developed the nanotags in the laboratory of Bruce Armitage, associate professor of chemistry in the Mellon College of Science (MCS) at Carnegie Mellon. "We’ve put it all into a very small package, which will allow us to detect molecules with great sensitivity without interfering with the biological processes we are trying to understand."
The high brightness of the nanotags should be of great help in detecting rare cancer cells within tissue biopsies, for example, which is important in determining whether treatments have been successful or if recurrence is likely, according to Armitage. In addition, DNA nanotags offer the opportunity to perform multicolor experiments. This feature is extremely useful for imaging applications, Armitage says, because the multiple colors can be seen simultaneously, requiring only one experiment using one laser and one fluorescence-imaging machine.
"For example, two different populations of cells, one healthy and the other cancerous, could be distinguished based on labeling them with different color fluorescent nanotags," Armitage said.
Benvin, Armitage and colleagues at Carnegie Mellon’s Molecular Biosensor and Imaging Center modeled their DNA nanotags on the structure of phycobiliproteins. Found in certain types of algae, such as the red and blue algae in fresh and marine waters, these proteins contain multiple, fluorescent pigments that work together to absorb light energy that is then transferred to chlorophyll, where it is used for photosynthesis. The Carnegie Mellon team has mimicked this efficient light-harvesting process in the design of their DNA nanotags to create incredibly bright, fluorescent labels.
"The primary advantages of our system are the simplicity of its design combined with the ease with which the fluorescence brightness and color can be tuned," Armitage said.
To achieve greater brightness, the Carnegie Mellon team assembled well-defined nanostructured DNA templates that bind multiple fluorescent dye molecules between base pairs in the DNA helix (see image). This arrangement keeps dyes far enough away from each other to avoid canceling out each other’s fluorescence. The DNA templates can also be modified to bind to other molecules or to the surface of a cell of interest. The innovative design creates nanotags with large light-harvesting capabilities and very high light-emission (fluorescence) intensities. Because the DNA can accommodate one dye for every two base pairs, a DNA nanostructure with 30 base pairs can bind up to 15 fluorescent dye molecules. The resulting dye-DNA complexes are approximately 15 times brighter than an individual dye molecule. And they can be made even brighter by simply increasing the number of base pairs in the DNA nanostructure.
Multicolor experiments are possible because the DNA nanotags contain "light-harvesting" dyes within the DNA helix that are excited by one wavelength of light and then transfer that excitation energy to "light-emitting" dyes on the nanotag’s surface. The light-emitting dyes can fluoresce at a different color from the light-harvesting dye. For example, one type of DNA nanotag can act as an antenna that efficiently harvests blue light and transfers that light energy to another dye within the nanostructure. The second dye then emits orange, red or even infrared light. Changing the light-harvesting dyes allows even more variation in the fluorescence color, Armitage said.
The nanotags are easily assembled by mixing commercially available DNA strands and fluorescent dyes. And while the work described by the Carnegie Mellon team relied on a relatively simple two-dimensional DNA nanostructure, Armitage notes that the rapidly growing field of DNA nanotechnology is generating increasingly intricate three-dimensional nanostructures that should lead to further improvements in brightness.
"We really feel that this is the tip of the iceberg and that nanotags 100 times brighter than existing labels can be developed in any color," he said.
Lauren Ward | EurekAlert!
Nanoparticle Exposure Can Awaken Dormant Viruses in the Lungs
16.01.2017 | Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt
Cholera bacteria infect more effectively with a simple twist of shape
13.01.2017 | Princeton University
Among the general public, solar thermal energy is currently associated with dark blue, rectangular collectors on building roofs. Technologies are needed for aesthetically high quality architecture which offer the architect more room for manoeuvre when it comes to low- and plus-energy buildings. With the “ArKol” project, researchers at Fraunhofer ISE together with partners are currently developing two façade collectors for solar thermal energy generation, which permit a high degree of design flexibility: a strip collector for opaque façade sections and a solar thermal blind for transparent sections. The current state of the two developments will be presented at the BAU 2017 trade fair.
As part of the “ArKol – development of architecturally highly integrated façade collectors with heat pipes” project, Fraunhofer ISE together with its partners...
At TU Wien, an alternative for resource intensive formwork for the construction of concrete domes was developed. It is now used in a test dome for the Austrian Federal Railways Infrastructure (ÖBB Infrastruktur).
Concrete shells are efficient structures, but not very resource efficient. The formwork for the construction of concrete domes alone requires a high amount of...
Many pathogens use certain sugar compounds from their host to help conceal themselves against the immune system. Scientists at the University of Bonn have now, in cooperation with researchers at the University of York in the United Kingdom, analyzed the dynamics of a bacterial molecule that is involved in this process. They demonstrate that the protein grabs onto the sugar molecule with a Pac Man-like chewing motion and holds it until it can be used. Their results could help design therapeutics that could make the protein poorer at grabbing and holding and hence compromise the pathogen in the host. The study has now been published in “Biophysical Journal”.
The cells of the mouth, nose and intestinal mucosa produce large quantities of a chemical called sialic acid. Many bacteria possess a special transport system...
UMD, NOAA collaboration demonstrates suitability of in-orbit datasets for weather satellite calibration
"Traffic and weather, together on the hour!" blasts your local radio station, while your smartphone knows the weather halfway across the world. A network of...
Fiber-reinforced plastics (FRP) are frequently used in the aeronautic and automobile industry. However, the repair of workpieces made of these composite materials is often less profitable than exchanging the part. In order to increase the lifetime of FRP parts and to make them more eco-efficient, the Laser Zentrum Hannover e.V. (LZH) and the Apodius GmbH want to combine a new measuring device for fiber layer orientation with an innovative laser-based repair process.
Defects in FRP pieces may be production or operation-related. Whether or not repair is cost-effective depends on the geometry of the defective area, the tools...
10.01.2017 | Event News
09.01.2017 | Event News
05.01.2017 | Event News
16.01.2017 | Power and Electrical Engineering
16.01.2017 | Information Technology
16.01.2017 | Power and Electrical Engineering