Novel microscopy generates new view of fuel cells

A novel microscopy method at the Department of Energy's Oak Ridge National Laboratory is helping scientists probe the reactions that limit widespread deployment of fuel cell technologies.

ORNL researchers applied a technique called electrochemical strain microscopy that enables them to examine the dynamics of oxygen reduction/evolution reactions in fuel cell materials, which may reveal ways to redesign or cut the costs of the energy devices. The team's findings were published in Nature Chemistry.

"If we can find a way to understand the operation of the fuel cell on the basic elementary level and determine what will make it work in the most optimum fashion, it would create an entirely new window of opportunity for the development of better materials and devices," said co-author Amit Kumar, a research scientist at ORNL's Center for Nanophase Materials Sciences.

Although fuel cells have long been touted as a highly efficient way to convert chemical energy into electrical energy, their high cost -- in large part due to the use of platinum as a catalyst -- has constrained commercial production and consumption.

Large amounts of platinum are used to catalyze the fuel cell's key reaction -- -the oxygen-reduction reaction, which controls the efficiency and longevity of the cell. Yet exactly how and where the reaction takes place had not been probed because existing device-level electrochemical techniques are ill suited to study the reaction at the nanoscale. ORNL co-author Sergei Kalinin explains that certain methods like electron microscopy had failed to capture the dynamics of fuel cell operation because their resolution was effectively too high.

"When you want to understand how a fuel cell works, you are not interested in where single atoms are, you're interested in how they move in nanometer scale volumes," Kalinin said. "The mobile ions in these solids behave almost like a liquid. They don't stay in place. The faster these mobile ions move, the better the material is for a fuel cell application. Electrochemical strain microscopy is able to image this ion mobility."

Other electrochemical techniques are unable to study oxygen-reduction reactions because they are limited to resolutions of 10's of microns -- 10,000 times larger than a nanometer.

"If the reaction is controlled by microstructure features that are much finer than a micron, let's say grain boundaries or single extended defects that are affecting the reaction, then you will never be able to catch what is giving rise to reduced or enhanced functionality of the fuel cell," said ORNL's Stephen Jesse, builder of the ESM microscope. "You would like to do this probing on a scale where you can identify each of these defects and correlate the functionality of the cell with these defects."

Although this study mainly focuses on the introduction of a technique, researchers explain their approach as a much-needed bridge between a theoretical and applied understanding of fuel cells.

"There is a huge gap between fundamental science and applied science for energy-related devices like fuel cells and batteries," Kalinin said. "The semiconducting industry, for example, is developing exponentially because the link between application and basic science is very well established. This is not the case in energy systems. They are usually much more complicated than semiconductors and therefore a lot of development is driven by trial and error type of work."

Co-authors on the study are University of Heidelberg's Francesco Ciucci and Anna Morozovska from the National Academy of Science of Ukraine, whose theoretical analysis was critical in explaining the ESM measurements.

Story Source:

The above story is reprinted (with editorial adaptations) from materials provided by DOE/Oak Ridge National Laboratory.

Journal Reference:

Amit Kumar, Francesco Ciucci, Anna N. Morozovska, Sergei V. Kalinin, Stephen Jesse. Measuring oxygen reduction/evolution reactions on the nanoscale. Nature Chemistry, 2011; DOI: 10.1038/nchem.1112

Revealing the inner workings of cells with newly developed Raman microscopy technique

Living cells are virtuosos of chemistry. At any one time, countless chemical reactions are taking place within each cell. For researchers trying to understand how cells function, unraveling this complex chemistry is an ongoing challenge. The process, however, could soon become a little more straightforward. Mikiko Sodeoka and colleagues at the RIKEN Advanced Science Institute at Wako, in collaboration with a team led by Katsumasa Fujita and Satoshi Kawata at Osaka University, have demonstrated how to tag molecules in a way that promises to be much more versatile than current methods.

Traditionally, researchers looking to study the role of a particular small molecule within a cell have tagged it with a fluorescent marker. Using a fluorescence microscope, the tagged substance can be followed as it moves around the cell. However, fluorescent tags are bulky, and so can disrupt the molecule’s normal cellular interactions. To get around this problem, molecules can sometimes be tagged after reaching their destination within the cell, but this technique only works in a limited number of cases.

Sodeoka and her team have now shown that a simple chemical substituent called an alkyne, which consists of just two carbon atoms joined together by a triple bond, can replace the bulky fluorescent tag. Their imaging technique relies on the fact that alkynes scatter a particular wavelength of light when irradiated with a laser—a process known as Raman scattering—which can be detected using a Raman microscope. No other cellular components scatter light at this wavelength, giving a clear picture of the molecule within the cell.

To demonstrate the potential of their technique, the researchers used an alkyne-tagged component of DNA known as EdU. They then used a Raman microscope developed by Fujita and his team to follow a group of replicating cells as they incorporated EdU into their DNA (Fig. 1). The technique took some work to optimize, says Sodeoka. “We were very happy when we could finally see the time-series pictures of the incorporation of EdU into DNA.”

The EdU experiment is just a proof of principle, Sodeoka adds. “At this point, the sensitivity of the alkyne tag using Raman microscopy is lower than fluorescent imaging,” she says. To improve the sensitivity, the team is working to optimize the attachment of the alkyne tag, and also to improve the Raman microscope itself. “If the sensitivity problem is solved, Raman imaging using alkynes as a small tag could become a powerful tool,” she concludes.

More information: Yamakoshi, H., et al. Imaging of EdU, an alkyne-tagged cell proliferation probe, by Raman microscopy. Journal of the American Chemical Society 133, 6102–6105 (2011).

Provided by RIKEN (news : web)

Polarized microscopy technique shows new details of how proteins are arranged

Whether you're talking about genes, or neurons, or the workings of a virus, at the most fundamental level, biology is a matter of proteins. So understanding what protein complexes look like and how they operate is the key to figuring out what makes cells tick. By harnessing the unique properties of polarized light, Rockefeller scientists have now developed a new technique that can help deduce the orientation of specific proteins within the cell. By turning their instruments toward the nuclear pore complex, a huge cluster of proteins that serves as a gateway to a cell's nucleus, the scientists say they have filled in the gaps left by other techniques and made important new discoveries about how the complex works.

"Our new technique allows us to measure how components of large protein complexes are arranged in relation to one another," says Sandy Simon, head of the Laboratory of Cellular Biophysics. "This has the potential to give us important new information about how the nuclear pore complex functions, but we believe it can also be applied to other multi-protein complexes such as those involved in DNA transcription, protein synthesis or viral replication."

Although researchers have spent years studying the workings of the nuclear pore complex, there is still much that has remained mysterious. One problem is that there is a "resolution gap" between the two techniques primarily used to visualize protein complexes. Electron microscopy can reveal the broad outlines of a large protein complex, but it can't show details. X-ray crystallography, meanwhile, can show minute detail but only of a small piece of the complex; it can't say how the individual pieces fit together. To further complicate matters, both techniques require fixed samples – while they can give you an idea of what something looks like at a moment in time, they can't tell you how its pieces might move.

The new technique was developed by Simon along with postdoc Alexa Mattheyses, graduate student Claire Atkinson and Martin Kampmann, a former a member of Günter Blobel's Laboratory of Cell Biology who is currently at the University of California, San Francisco. It takes advantage of the properties of polarized light to show how specific proteins are aligned in relation to one another. After genetically attaching fluorescent markers to individual components of the nuclear pore complex, the scientists replaced the cell's own copy of the gene that encodes the protein with the new form that has the fluorescent tag. Then, they used customized microscopes to measure the orientation of the waves of light the fluorescently tagged proteins emitted. By combining these measurements with known data about the structure of the complex, the scientists can confirm or deny the accuracy of previously suggested models.

"Our experimental approach to the structure is synergistic with other studies being conducted at Rockefeller, including analysis with X-ray crystallography in Günter's lab and electron microscopy and computer analysis in Mike Rout's lab," says Simon. "By utilizing multiple techniques, we are able to get a more precise picture of these complexes than has ever been possible before."

The scientists used the technique to study nuclear pore complexes in both budding yeast and human cells. In the case of the human cells, their new data shows that multiple copies of a key building block of the nuclear pore complex, the Y-shaped subcomplex, are arranged head-to-tail, rather than like fence posts, confirming a model proposed by Blobel in 2007.

"As a graduate student with Günter Blobel, I determined the three-dimensional structure of the Y-shaped subcomplex using electron microscopy," says Kampmann. "However, it was still a mystery how these 'Y's are arranged. The new technique we have developed reveals the orientation of building blocks in the cell, and we hope that it will eventually enable us to assemble individual crystal structures into a high-resolution map of the entire nuclear pore complex."

Eventually, the scientists say their technique could go even further. Because the proteins' fluorescence can be measured while the cells are still alive, it could give scientists new insights into how protein complexes react to varying environmental conditions, and how their configurations change over time.

"What happens when other proteins pass through the nuclear pore? Does the orientation of the nucleoporins change? With this technique, can find out not only what the pore looks like when it's sitting still, but what happens to it when it's active," Simon says. Their first characterization of the dynamics of the nuclear pore proteins was published recently in The Biophysical Journal.

Provided by Rockefeller University (news : web)

Electron microscopy: New type of genetic tag illuminates life in never-before-seen detail

By modifying a protein from a plant that is much favored by science, researchers at the University of California, San Diego School of Medicine and colleagues have created a new type of genetic tag visible under an electron microscope, illuminating life in never-before-seen detail.

Led by Nobel laureate Roger Tsien, PhD, Howard Hughes Medical Institute investigator and UCSD professor of pharmacology, chemistry and biochemistry, a team of scientists radically re-engineered a light-absorbing protein from the flowering cress plant Arabidopsis thaliana. When exposed to blue light, the altered protein produces abundant singlet oxygen, a form of molecular oxygen that can be made visible by electron microscopy (EM).

The findings are published in the online, open access journal PLoS Biology.

Tsien was co-winner of the 2008 Nobel Prize in chemistry for his role in helping develop and expand the use of green fluorescent protein (GFP), a protein from jellyfish that is now widely employed in light microscopy to peer inside living cells or whole animals and observe molecules interacting in real-time. Tsien said the development of the small, highly engineered Arabidopsis protein, dubbed "miniSOG," may elevate the abilities of electron microscopy in the same way that GFP and its relatives have made modern light microscopy in biological research much more powerful and useful.

"The big advantage of EM is that it has much higher spatial resolution than light microscopy. You can get up to a hundred-fold higher useful magnification from EM than from light microscopy," said Tsien. The result has been extraordinarily detailed, three-dimensional images of microscopic objects at resolutions measuring in the tens of nanometers, tiny enough to meticulously render the internal anatomy of individual cells. But current EM technologies do not distinguish or highlight individual proteins in these images. Although individual proteins can be tagged with GFP or other fluorescent proteins to aid localization by light microscopy, there has been no equivalent technology for the higher-resolution images provided by EM.

To create this ability, the scientists began with a protein from Arabidopsis that absorbs incoming blue light. It's normal function is to trigger biochemical signals that inform the plant how much sunlight it is receiving. "We rationally engineered the protein based on its atomic model so that it changes incoming blue light into a little bit of green fluorescence and a lot of singlet oxygen," said the paper's first author, Xiaokun Shu, now an assistant professor at UC San Francisco. Established methods were then used to convert singlet oxygen production into a tissue stain that the electron microscope can "see." The scientists tested the modified protein's utility as an EM marker by first using it to confirm the locations of several well-understood proteins in mammalian cells, nematodes and rodents, and then used miniSOG to successfully tag two neuronal proteins in mice whose locations had not been known.

Tsien is optimistic that miniSOG will grant new powers to electron microscopy, permitting scientists to pursue answers to questions previously impossible to ask. MiniSOG will especially be useful to scientists who investigate cellular and subcellular structures including neuronal circuits at nanometer resolution in multicellular organisms since previous methods have great difficulty in achieving both efficient labeling and good preservation of the structures under study. While EM can provide much higher useful magnification than light microscopy, EM will not replace light microscopy. "When we use miniSOG, we see the tagged proteins plus the landmarks that we are used to navigating by," said Tsien. "On the other hand, EM has the disadvantage that it gives a snapshot of cells before we killed them (to make the image), whereas light microscopy can show the dynamics in live cells. Each technique has different complementary strengths and weaknesses."

Co-authors of the paper include: Varda Lev-Ram, UCSD Department of Pharmacology; Thomas J. Deerinck, National Center for Microscopy and Imaging Research, Center for Research on Biological Systems, UCSD; Yingchuan Qi and Yishi Jin, Howard Hughes Medical Institute, UCSD and UCSD Division of Biological Science; Ericka B. Ramko and Michael W. Davidson, National High Magnetic Field Laboratory and Department of Biological Science, Florida State University; and Mark H. Ellisman, National Center for Microscopy and Imaging Research, Center for Research on Biological Systems, UCSD and UCSD Department of Neurosciences

Story Source:

The above story is reprinted  from materials provided by Public Library of Science, via EurekAlert!, a service of AAAS.

Journal Reference:

Xiaokun Shu, Varda Lev-Ram, Thomas J. Deerinck, Yingchuan Qi, Ericka B. Ramko, Michael W. Davidson, Yishi Jin, Mark H. Ellisman, Roger Y. Tsien. A Genetically Encoded Tag for Correlated Light and Electron Microscopy of Intact Cells, Tissues, and Organisms. PLoS Biology, 2011; 9 (4): e1001041 DOI: 10.1371/journal.pbio.1001041

GRIN plasmonics: A practical path to superfast computing, ultrapowerful optical microscopy and invisibility carpet-cloaking devices

They said it could be done and now they've done it. What's more, they did it with a GRIN. A team of researchers with the U.S. Department of Energy (DOE)'s Lawrence Berkeley National Laboratory (Berkeley Lab) and the University of California, Berkeley, have carried out the first experimental demonstration of GRIN -- for gradient index -- plasmonics, a hybrid technology that opens the door to a wide range of exotic optics, including superfast computers based on light rather than electronic signals, ultra-powerful optical microscopes able to resolve DNA molecules with visible light, and "invisibility" carpet-cloaking devices.

Working with composites featuring a dielectric (non-conducting) material on a metal substrate, and "grey-scale" electron beam lithography, a standard method in the computer chip industry for patterning 3-D surface topographies, the researchers have fabricated highly efficient plasmonic versions of Luneburg and Eaton lenses. A Luneburg lens focuses light from all directions equally well, and an Eaton lens bends light 90 degrees from all incoming directions.

"This past year, we used computer simulations to demonstrate that with only moderate modifications of an isotropic dielectric material in a dielectric-metal composite, it would be possible to achieve practical transformation optics results," says Xiang Zhang, who led this research. "Our GRIN plasmonics technique provides a practical way for routing light at very small scales and producing efficient functional plasmonic devices."

Zhang, a principal investigator with Berkeley Lab's Materials Sciences Division and director of UC Berkeley's Nano-scale Science and Engineering Center (SINAM), is the corresponding author of a paper in the journal Nature Nanotechnology, describing this work titled, "Plasmonic Luneburg and Eaton Lenses." Co-authoring the paper were Thomas Zentgraf, Yongmin Liu, Maiken Mikkelsen and Jason Valentine.

GRIN plasmonics combines methodologies from transformation optics and plasmonics, two rising new fields of science that could revolutionize what we are able to do with light. In transformation optics, the physical space through which light travels is warped to control the light's trajectory, similar to the way in which outer space is warped by a massive object under Einstein's relativity theory. In plasmonics, light is confined in dimensions smaller than the wavelength of photons in free space, making it possible to match the different length-scales associated with photonics and electronics in a single nanoscale device.

"Applying transformation optics to plasmonics allows for precise control of strongly confined light waves in the context of two-dimensional optics," Zhang says. "Our technique is analogous to the well-known GRIN optics technique, whereas previous plasmonic techniques were realized by discrete structuring of the metal surface in a metal-dielectric composite."

Like all plasmonic technologies, GRIN plasmonics starts with an electronic surface wave that rolls through the conduction electrons on a metal. Just as the energy in a wave of light is carried in a quantized particle-like unit called a photon, so, too, is plasmonic energy carried in a quasi-particle called a plasmon. Plasmons will interact with photons at the interface of a metal and dielectric to form yet another quasi-particle, a surface plasmon polariton (SPP).

The Luneburg and Eaton lenses fabricated by Zhang and his co-authors interacted with SPPs rather than photons. To make these lenses, the researchers worked with a thin dielectric film (a thermplastic called PMMA) on top of a gold surface. When applying grey-scale electron beam lithography, the researchers exposed the dielectric film to an electron beam that was varied in dosage (charge per unit area) as it moved across the film's surface. This resulted in highly controlled differences in film thickness across the length of the dielectric that altered the local propagation of SPPs. In turn, the "mode index," which determines how fast the SPPs will propagate, is altered so that the direction of the SPPs can be influenced.

"By adiabatically tailoring the topology of the dielectric layer adjacent to the metal surface, we're able to continuously modify the mode index of SPPs," says Zentgraf. "As a result, we can manipulate the flow of SPPs with a greater degree of freedom in the context of two-dimensional optics."

Says Liu, "The practicality of working only with the purely dielectric material to transform SPPs is a big selling point for GRIN plasmonics. Controlling the physical properties of metals on the nanometer length-scale, which is the penetration depth of electromagnetic waves associated with SPPs extending below the metal surfaces, is beyond the reach of existing nanofabrication techniques."

Adds Zentgraf, "Our approach has the potential to achieve low-loss functional plasmonic elements with a standard fabrication technology that is fully compatible with active plasmonics."

In the Nature Nanotechnology paper, the researchers say that inefficiencies in plasmonic devices due to SPPs lost through scattering could be reduced even further by incorporating various SPP gain materials, such as fluorescent dye molecules, directly into the dielectric. This, they say, would lead to an increased propagation distance that is highly desired for optical and plasmonic devices. It should also enable the realization of two-dimensional plasmonic elements beyond the Luneburg and Eaton lenses.

Says Mikkelsen, "GRIN plasmonics can be immediately applied to the design and production of various plasmonic elements, such as waveguides and beam splitters, to improve the performance of integrated plasmonics. Currently we are working on more complex, transformational plasmonic devices, such as plasmonic collimators, single plasmonic elements with multiple functions, and plasmonic lenses with enhanced performance."

This research was supported by the U.S. Army Research Office and the National Science Foundation's Nano-scale Science and Engineering Center.

Story Source:

The above story is reprinted (with editorial adaptations by ScienceDaily staff) from materials provided by DOE/Lawrence Berkeley National Laboratory.

Journal Reference:

Thomas Zentgraf, Yongmin Liu, Maiken H. Mikkelsen, Jason Valentine, Xiang Zhang. Plasmonic Luneburg and Eaton lenses. Nature Nanotechnology, 2011; DOI: 10.1038/nnano.2010.282