National Center for Microscopy and Imaging Research

San Diego, CA, United States

National Center for Microscopy and Imaging Research

San Diego, CA, United States
SEARCH FILTERS
Time filter
Source Type

News Article | April 20, 2017
Site: news.yahoo.com

This undated microscope image made available by the National Center for Microscopy and Imaging Research shows HeLa cells. Until these cells came along, whenever human cells were put in a lab dish, they would die immediately or reproduce only a few times. Henrietta Lacks' cells, by contrast, grew indefinitely. They were "perpetual, everlasting, death-defying, or whatever other word you want to use to describe immortal," says Dr. Francis Collins, director of the U.S. National Institutes of Health. (National Center for Microscopy and Imaging Research via AP) NEW YORK (AP) — What happened in the 1951 case of Henrietta Lacks, and could it happen again today? The story of the woman who unwittingly spurred a scientific bonanza made for a best-selling book in 2010. On Saturday, it returns in an HBO film with Oprah Winfrey portraying Lacks' daughter Deborah. Cells taken from Henrietta Lacks have been widely used in biomedical research. They came from a tumor sample taken from Lacks — who never gave permission for their use. A look at the case: HOW DID DOCTORS GET THE CELLS? As the book relates, Lacks was under anesthesia on an operating table at Johns Hopkins Hospital in Baltimore one day in 1951, undergoing treatment for cervical cancer. A hospital researcher had been collecting cervical cancer cells to see if they would grow continuously in the laboratory. So the surgeon treating Lacks shaved a dime-sized piece of tissue from her tumor for that project. Nobody had asked Lacks if she wanted to provide cells for the research. She died later that year. WAS IT ILLEGAL TO TAKE THE CELLS WITHOUT HER PERMISSION? Not at that time. "What happened to Henrietta Lacks was commonly done," says bioethicist Dr. Robert Klitzman of Columbia University in New York. WHAT ARE THE RULES NOW IN THE U.S.? Specimens intended specifically for research can be collected only if the donor gives consent first. If cells or tissues are instead removed for diagnosis and treatment, that is considered part of the patient's general consent for treatment. But there's a twist. Once a specimen is no longer needed for treating the patient and would otherwise be discarded, scientists can use it for research. No further consent is needed, as along as information identifying the patient as the source is removed and the specimen can't be traced back to the patient, says Johns Hopkins University bioethicist Jeffrey Kahn. IF A SPECIMEN LEADS TO A PRODUCT, DOES THE DONOR HAVE A RIGHT TO SHARE IN THE PROFITS? Generally not, because the consent form for donation or treatment usually waives any such legal right. WHAT WAS SO SPECIAL ABOUT LACKS' CELLS? Until they came along, whenever human cells were put in a lab dish, they would die immediately or reproduce only a few times. Her cells, by contrast, could be grown indefinitely. They were "perpetual, everlasting, death-defying, or whatever other word you want to use to describe immortal," as Dr. Francis Collins, director of the U.S. National Institutes of Health, put it. So they provided an unprecedented stock of human cells that could be shipped worldwide for experiments. They quickly became the most popular human cells for research, and have been cited in more than 74,000 scientific publications. HOW HAVE RESEARCHERS USED THE CELLS? The so-called "HeLa" cells became crucial for key developments in such areas as basic biology, understanding viruses and other germs, cancer treatments, in vitro fertilization and development of vaccines, including the polio vaccine. WHAT MAKES THEM GROW SO WELL? Researchers proposed a possible answer in 2013. Virtually all cases of cervical cancer are caused by infection with human papillomavirus , which inserts its genetic material into a human cell's DNA. Scientists who examined the DNA of HeLa cells suggested that happened in a place that strongly activated a cancer-promoting gene. That might explain both why Lacks' cancer was so aggressive and why the cells grow so robustly in a lab dish. DID EVERYBODY ALWAYS KNOW THE ORIGIN OF THE CELLS? No. Lacks was named publicly only in 1971, by an article in a medical journal. Her story appeared in some magazines in the 1970s, and in a 1997 documentary on BBC. She became famous in 2010 with publication of Rebecca Skloot's best-selling book, "The Immortal Life of Henrietta Lacks." Follow Malcolm Ritter at http://twitter.com/malcolmritter His recent work can be found at http://tinyurl.com/RitterAP


Boassa D.,National Center for Microscopy and Imaging Research | Berlanga M.L.,National Center for Microscopy and Imaging Research | Yang M.A.,Urbana University | Yang M.A.,Concordia University at Saint Paul | And 8 more authors.
Journal of Neuroscience | Year: 2013

Modifications to the gene encoding human α-synuclein have been linked to the development of Parkinson's disease. The highly conserved structure of α-synuclein suggests a functional interaction with membranes, and several lines of evidence point to a role in vesicle-related processes within nerve terminals. Using recombinant fusions of humanα-synuclein, including new genetic tags developed for correlated light microscopy and electron microscopy (the tetracysteine-biarsenical labeling system or the new fluorescent protein for electron microscopy, MiniSOG), we determined the distribution ofα-synuclein when overexpressed in primaryneuronsat supramolecular and cellular scales in three dimensions (3D). We observed specific association of α-synuclein with a large and otherwise poorly characterized membranous organelle system of the presynaptic terminal, as well as with smaller vesicular structures within these boutons. Furthermore, α-synuclein was localized to multiple elements of the protein degradation pathway, including multivesicular bodies in the axons and lysosomes within neuronal cell bodies. Examination of synapses in brains of transgenic mice overexpressing human α-synuclein revealed alterations of the presynaptic endomembrane systems similar to our findings in cell culture. Three-dimensional electron tomographic analysis of enlarged presynaptic terminals in several brain areas revealed that these terminals were filled withmembrane-boundedorganelles, including tubulovesicular structures similar to whatweobserved in vitro. We propose that α-synuclein overexpression is associated with hypertrophy of membrane systems of the presynaptic terminal previously shown to have arole in vesicle recycling. Our data support the conclusion that α-synuclein is involved in processes associated with the sorting, channeling, packaging, and transport of synaptic material destined for degradation. © 2013 the authors.


News Article | November 3, 2016
Site: www.eurekalert.org

The best microscope we have for peering inside of a cell can now produce color images. University of California, San Diego, scientists demonstrate this advancement in electron microscopy -- of the ability to magnify objects up to ten million times -- with photographs of cellular membranes and the synaptic connections between brain cells. The development of multicolor electron microscopy, presented November 3 in Cell Chemical Biology, was jointly overseen by Mark Ellisman and the late Roger Tsien, a 2008 Chemistry Nobel Prize laureate and visionary for cellular imaging who died unexpectedly over the summer. With their new method, which the research team worked on for nearly 15 years, up to three colors at a time (green, red, or yellow) can be used in an image. A detector on the microscope captures electrons lost from metal ions painted over the specimen and records the metal's energy loss signature as a color. A technician must add the ionized metals one at a time and then lay the full color map over the still microscopy image. "It's a bit like when you first see a color photograph after having only known black and white -- for the last 50 years or so, we've been so used to monochrome electron micrographs that it's now hard to imagine that we could go back," says first author Stephen Adams, a UCSD chemist. "This method has many potential applications in biology; in the paper, we demonstrate how it can distinguish cellular compartments or track proteins and tag cells." For the multicolor effect to work, the researchers needed metal complexes that are stable enough to withstand application (meaning they don't quickly deteriorate and blur the image) and have a distinct electron energy loss signature. The researchers used ionized lanthanum (La), cerium (Ce), and praseodymium (Pr) -- all metals in the lanthanide family -- with each metal complex laid down sequentially as a precipitate onto the specimen as it sits in the microscope. "One challenge that kept us from publishing this much earlier, because we had the chemistry and we had an instrument that worked about 4 years ago, was we needed a way to deposit the metal compounds sequentially," says co-senior author Mark Ellisman, director of the National Center for Microscopy and Imaging Research at UCSD. "We spent an awful lot of time trying to figure out how to deposit one of the lanthanides and then clear it so that it didn't react when we deposited a second signal on the first site." Once the application process had been established, the research team illustrated the power of multicolor electron microscopy by visualizing two brain cells sharing a single synapse. They also show peptides entering through a cell membrane. The new method is analogous to fluorescence microscopy--a tool that detects colored light emitted from glowing proteins tagged in a biological specimen--but benefits from the details that can only be captured by electron microscopy. Notably, this paper is one of the last that Roger Tsien, who won a 2008 Nobel Prize in Chemistry for the discovery and application of green fluorescent protein to biochemical imaging, saw accepted by a journal before his death last August. He did the first experiments to develop the chemical compounds needed for the multicolor imaging method nearly 15 years ago. As a Christmas present to himself, he would spend 2 weeks at the bench, and this was one of his holiday projects. "One theme that has gone through all of Roger's work is the desire to peer more closely into the workings of the cell," Adams says. "With all of the fluorescence techniques that he's introduced, he was able to do that in live cells, and make action movies of them in vivid colors. But he always wanted to look closer, and now he's left the beginnings for a method where we can add colors to electron microscopy." "This is clearly an example of Roger's brilliance at chemistry and how he saw that if we could do this, we would be able to enjoy the advantages of electron microscopy," adds Ellisman, a longtime collaborator who was co-senior author with Tsien on dozens of studies. "The biggest advantage of electron microscopy that we saw is that you have weak contrasts by the nature of the way that staining works so color-specific label give context to all of the rich information in the scene of which molecules are operating." The researchers say there is more chemistry to be done to perfect the metal ion application process as well as produce images with three or more colors. There may also be ways to increase the amount of metal ions that can be deposited, which could help with resolution. Many in the biochemical community should be able to begin using this technique right away, as it takes advantage of tools that are already found in laboratories. This work was supported by UCSD Graduate Training Programs in Cellular and Molecular Pharmacology and in Neuroplasticity of Aging, the National Institutes of Health, and the W.M. Keck Foundation. Cell Chemical Biology, Adams et al.: "Multicolor electron microscopy for simultaneous visualization of multiple molecular species" http://www. (16)30357-9/10.1016 Cell Chemical Biology (@CellChemBiol), published by Cell Press, is a monthly journal publishing research and review content of exceptional interest for the chemical biology community. The journal's mission is to support and promote chemical biology and drive conversation and collaboration between chemical and life sciences. For more information, please visit http://www. . To receive Cell Press media alerts, contact press@cell.com.


« SLAC, U Toronto team develops new highly efficient ternary OER catalyst for water-splitting using earth-abundant metals; >3x TOF prior record-holder | Main | Six automated truck platoons to compete in European Truck Platooning Challenge » Researchers from the J. Craig Venter Institute (JCVI) and Synthetic Genomics, Inc. (SGI) have designed and constructed of the first minimal synthetic bacterial cell, JCVI-syn3.0. Using the first synthetic cell, Mycoplasma mycoides JCVI-syn1.0 (created by this same team in 2010, earlier post), JCVI-syn3.0 was developed through a design, build, and test process using genes from JCVI-syn1.0. The new minimal synthetic cell contains 531,560 base pairs and just 473 genes, making it the smallest genome of any organism that can be grown in laboratory media. Of these genes 149 are of unknown biological function. By comparison the first synthetic cell, M. mycoides JCVI-syn1.0 has 1.08 million base pairs and 901 genes. A paper describing this research is being published in the journal Science by lead authors Clyde A. Hutchison, III, Ph.D. and Ray-Yuan Chuang, Ph.D., senior author J. Craig Venter, Ph.D., and senior team of Hamilton O. Smith, MD, Daniel G. Gibson, Ph.D., and John I. Glass, Ph.D. Our attempt to design and create a new species, while ultimately successful, revealed that 32% of the genes essential for life in this cell are of unknown function, and showed that many are highly conserved in numerous species. All the bioinformatics studies over the past 20 years have underestimated the number of essential genes by focusing only on the known world. This is an important observation that we are carrying forward into the study of the human genome. The research to construct the first minimal synthetic cell at JCVI was the culmination of 20 years of research that began in 1995 after the genome sequencing of the first free-living organism, Haemophilus influenza, followed by the sequencing of Mycoplasma genitalium. A comparison of these two genomes revealed a common set of 256 genes which the team thought could be a minimal set of genes needed for viability. In 1999 Dr. Hutchison led a team who published a paper describing the use of global transposon mutagenesis techniques to identify the nonessential genes in M. genitalium. Over the last 50 years more than 2,000 publications have contemplated minimal cells and their use in elucidating first principals of biology. From the start, the goal of the JCVI team was similar—build a minimal operating system of a cell to understand biology but to also have a desirable chassis for use in industrial applications. The creation of the first synthetic cell in 2010 did not inform new genome design principles since the M. mycoides genome was mostly recapitulated as in nature. Rather, it established a work flow for building and testing whole genome designs, including a minimal cell, from the bottom up starting from a genome sequence. To create JCVI-syn3.0, the team used an approach of whole genome design and chemical synthesis followed by genome transplantation to test if the cell was viable. Their first attempt to minimize the genome began with a simple approach using information in the biochemical literature and some limited transposon mutagenesis work, but this did not result in a viable genome. After improving transposon methods, they discovered a set of quasi-essential genes that are necessary for robust growth which explained the failure of their first attempt. To facilitate debugging of non-functional reduced genome segments, the team built the genome in eight segments at a time so that each could be tested separately before combining them to generate a minimal genome. The team also explored gene order and how that affects cell growth and viability, noting that gene content was more critical to cell viability than gene order. They went through three cycles of designing, building, and testing ensuring that the quasi-essential genes remained, which in the end resulted in a viable, self-replicating minimal synthetic cell that contained just 473 genes, 35 of which are RNA-coding. In addition, the cell contains a unique 16S gene sequence. The team was able to assign biological function to the majority of the genes with 41% of them responsible for genome expression information, 18% related to cell membrane structure and function, 17% related to cytosolic metabolism, and 7% preservation of genome information. However, a surprising 149 genes could not be assigned a specific biological function despite intensive study. This remains an area of continued work for the researchers. The team concludes that a major outcome of this minimal cell program are new tools and semi-automated processes for whole genome synthesis. Many of these synthetic biology tools and services are commercially available through SGI and SGI-DNA including a synthetic DNA construction service specializing in building large and complex DNA fragments including combinatorial gene libraries, Archetype genomics software, Gibson Assembly kits, and the BioXp, which is a benchtop instrument for producing accurate synthetic DNA fragments. Other authors on the paper are: Thomas J. Deerinck and Mark H. Ellisman, Ph.D., University of California, San Diego National Center for Microscopy and Imaging Research; James F. Pelletier, Center for Bits and Atoms and Department of Physics, Massachusetts Institute of Technology; Elizabeth A. Strychalski, National Institute of Standards and Technology. This work was funded by SGI, the JCVI endowment and the Defense Advanced Research Projects Agency’s Living Foundries program, HR0011-12-C-0063.


News Article | November 3, 2016
Site: www.rdmag.com

The best microscope we have for peering inside of a cell can now produce color images. University of California, San Diego, scientists demonstrate this advancement in electron microscopy -- of the ability to magnify objects up to ten million times -- with photographs of cellular membranes and the synaptic connections between brain cells. The development of multicolor electron microscopy, presented November 3 in Cell Chemical Biology, was jointly overseen by Mark Ellisman and the late Roger Tsien, a 2008 Chemistry Nobel Prize laureate and visionary for cellular imaging who died unexpectedly over the summer. With their new method, which the research team worked on for nearly 15 years, up to three colors at a time (green, red, or yellow) can be used in an image. A detector on the microscope captures electrons lost from metal ions painted over the specimen and records the metal's energy loss signature as a color. A technician must add the ionized metals one at a time and then lay the full color map over the still microscopy image. "It's a bit like when you first see a color photograph after having only known black and white -- for the last 50 years or so, we've been so used to monochrome electron micrographs that it's now hard to imagine that we could go back," says first author Stephen Adams, a UCSD chemist. "This method has many potential applications in biology; in the paper, we demonstrate how it can distinguish cellular compartments or track proteins and tag cells." For the multicolor effect to work, the researchers needed metal complexes that are stable enough to withstand application (meaning they don't quickly deteriorate and blur the image) and have a distinct electron energy loss signature. The researchers used ionized lanthanum (La), cerium (Ce), and praseodymium (Pr) -- all metals in the lanthanide family -- with each metal complex laid down sequentially as a precipitate onto the specimen as it sits in the microscope. "One challenge that kept us from publishing this much earlier, because we had the chemistry and we had an instrument that worked about 4 years ago, was we needed a way to deposit the metal compounds sequentially," says co-senior author Mark Ellisman, director of the National Center for Microscopy and Imaging Research at UCSD. "We spent an awful lot of time trying to figure out how to deposit one of the lanthanides and then clear it so that it didn't react when we deposited a second signal on the first site." Once the application process had been established, the research team illustrated the power of multicolor electron microscopy by visualizing two brain cells sharing a single synapse. They also show peptides entering through a cell membrane. The new method is analogous to fluorescence microscopy--a tool that detects colored light emitted from glowing proteins tagged in a biological specimen--but benefits from the details that can only be captured by electron microscopy. Notably, this paper is one of the last that Roger Tsien, who won a 2008 Nobel Prize in Chemistry for the discovery and application of green fluorescent protein to biochemical imaging, saw accepted by a journal before his death last August. He did the first experiments to develop the chemical compounds needed for the multicolor imaging method nearly 15 years ago. As a Christmas present to himself, he would spend 2 weeks at the bench, and this was one of his holiday projects. "One theme that has gone through all of Roger's work is the desire to peer more closely into the workings of the cell," Adams says. "With all of the fluorescence techniques that he's introduced, he was able to do that in live cells, and make action movies of them in vivid colors. But he always wanted to look closer, and now he's left the beginnings for a method where we can add colors to electron microscopy." "This is clearly an example of Roger's brilliance at chemistry and how he saw that if we could do this, we would be able to enjoy the advantages of electron microscopy," adds Ellisman, a longtime collaborator who was co-senior author with Tsien on dozens of studies. "The biggest advantage of electron microscopy that we saw is that you have weak contrasts by the nature of the way that staining works so color-specific label give context to all of the rich information in the scene of which molecules are operating." The researchers say there is more chemistry to be done to perfect the metal ion application process as well as produce images with three or more colors. There may also be ways to increase the amount of metal ions that can be deposited, which could help with resolution. Many in the biochemical community should be able to begin using this technique right away, as it takes advantage of tools that are already found in laboratories.


News Article | November 4, 2016
Site: www.biosciencetechnology.com

The best microscope we have for peering inside of a cell can now produce color images. University of California, San Diego, scientists demonstrate this advancement in electron microscopy -- of the ability to magnify objects up to ten million times -- with photographs of cellular membranes and the synaptic connections between brain cells. The development of multicolor electron microscopy, presented November 3 in Cell Chemical Biology, was jointly overseen by Mark Ellisman and the late Roger Tsien, a 2008 Chemistry Nobel Prize laureate and visionary for cellular imaging who died unexpectedly over the summer. With their new method, which the research team worked on for nearly 15 years, up to three colors at a time (green, red, or yellow) can be used in an image. A detector on the microscope captures electrons lost from metal ions painted over the specimen and records the metal's energy loss signature as a color. A technician must add the ionized metals one at a time and then lay the full color map over the still microscopy image. "It's a bit like when you first see a color photograph after having only known black and white -- for the last 50 years or so, we've been so used to monochrome electron micrographs that it's now hard to imagine that we could go back," says first author Stephen Adams, a UCSD chemist. "This method has many potential applications in biology; in the paper, we demonstrate how it can distinguish cellular compartments or track proteins and tag cells." For the multicolor effect to work, the researchers needed metal complexes that are stable enough to withstand application (meaning they don't quickly deteriorate and blur the image) and have a distinct electron energy loss signature. The researchers used ionized lanthanum (La), cerium (Ce), and praseodymium (Pr) -- all metals in the lanthanide family -- with each metal complex laid down sequentially as a precipitate onto the specimen as it sits in the microscope. "One challenge that kept us from publishing this much earlier, because we had the chemistry and we had an instrument that worked about 4 years ago, was we needed a way to deposit the metal compounds sequentially," says co-senior author Mark Ellisman, director of the National Center for Microscopy and Imaging Research at UCSD. "We spent an awful lot of time trying to figure out how to deposit one of the lanthanides and then clear it so that it didn't react when we deposited a second signal on the first site." Once the application process had been established, the research team illustrated the power of multicolor electron microscopy by visualizing two brain cells sharing a single synapse. They also show peptides entering through a cell membrane. The new method is analogous to fluorescence microscopy--a tool that detects colored light emitted from glowing proteins tagged in a biological specimen--but benefits from the details that can only be captured by electron microscopy. Notably, this paper is one of the last that Roger Tsien, who won a 2008 Nobel Prize in Chemistry for the discovery and application of green fluorescent protein to biochemical imaging, saw accepted by a journal before his death last August. He did the first experiments to develop the chemical compounds needed for the multicolor imaging method nearly 15 years ago. As a Christmas present to himself, he would spend 2 weeks at the bench, and this was one of his holiday projects. "One theme that has gone through all of Roger's work is the desire to peer more closely into the workings of the cell," Adams says. "With all of the fluorescence techniques that he's introduced, he was able to do that in live cells, and make action movies of them in vivid colors. But he always wanted to look closer, and now he's left the beginnings for a method where we can add colors to electron microscopy." "This is clearly an example of Roger's brilliance at chemistry and how he saw that if we could do this, we would be able to enjoy the advantages of electron microscopy," adds Ellisman, a longtime collaborator who was co-senior author with Tsien on dozens of studies. "The biggest advantage of electron microscopy that we saw is that you have weak contrasts by the nature of the way that staining works so color-specific label give context to all of the rich information in the scene of which molecules are operating." The researchers say there is more chemistry to be done to perfect the metal ion application process as well as produce images with three or more colors. There may also be ways to increase the amount of metal ions that can be deposited, which could help with resolution. Many in the biochemical community should be able to begin using this technique right away, as it takes advantage of tools that are already found in laboratories.


News Article | November 3, 2016
Site: www.eurekalert.org

Electron microscopy (EM), which uses particle beams of accelerated electrons to interrogate specimens, has long been a leading technology for revealing the shape and structure of the tiniest objects, from the cells which make up the bodies organs and microbes to individual building blocks or molecules which comprise cells, in often dramatic three-dimensional detail. But current EM techniques are limited in that they produce images only in grayscale, with colorization added later. In a paper published online November 3 in Cell Chemical Biology, researchers at University of California San Diego School of Medicine and Howard Hughes Medical Institute describe a new form of multicolor EM that allows for simultaneous visualization of multiple molecular species. "The ability to discern multiple specific molecules simultaneously adds a new dimension. It reveals details, actions and processes that aren't necessarily visible -- or even suspected -- in a more monochromatic view," said Mark H. Ellisman, PhD, professor in the Department of Neurosciences and director of the National Center for Microscopy and Imaging Research. Ellisman is a co-senior author of the study. Roger Tsien, PhD, professor of pharmacology, chemistry and biochemistry, was also co-senior author. Tsien, who passed away August 24, was co-winner of the 2008 Nobel Prize in chemistry for his work developing green fluorescent proteins as an imaging research tool. He was considered a leading light in microscopy and imaging research and was cited as one of the "world's most influential scientific minds" by Thomson Reuters earlier this year. Although there have been major improvements in multicolor and super-resolution fluorescence microscopy in recent years, comparable progress in EM has been more limited, achieved through automation and developments like the miniSOG protein, a new type of genetic tag visible under an EM microscope that was developed by Tsien, Ellisman and colleagues in 2011. To create multicolor EM images, first author and project scientist Stephen R. Adams, PhD, said researchers sequentially painted cellular structures such as proteins, membranes or whole cells with different "rare earth" metals, such as lanthanum, cerium and praseodymium in the form of precipitates. "A transmission electron microscope can distinguish each of these metals by electron energy-loss to give elemental maps of each that can be overlaid in color on the familiar monochrome electron micrograph," said Adams. "Each color highlights a different component of the cellular ultrastructure." Multicolor EM offers the possibility to differentiate detail not possible with standard EM, which uses gold particles to label structures but which appear in images as sometimes hard-to-distinguish black spots. It provides spatial resolution not possible with fluorescence microscopy. "This new method gives a more complete and easily detectable readout of the cellular components as colors," said Adams. "In theory, we should be able to add many more colors if we can develop more ways of precipitating additional lanthanides. The method is quite simple to do, uses easily made chemicals and requires detectors that are already present on many transmission electron microscopes so it is potentially readily transferable to other laboratories. Further research is needed to improve the chemistry and sensitivity of the method, but this work will hopefully inspire other groups to devise similar methods in this field." Co-authors include: Mason R. Mackey, Ranjan Ramachandra, Sakina F. Palida Lemieux, Eric A. Bushong, Margaret T. Butko, Ben N.G. Giepmans, and Paul Steinbach, all at UC San Diego at the time they contributed to the work.


News Article | November 4, 2016
Site: www.sciencedaily.com

Electron microscopy (EM), which uses particle beams of accelerated electrons to interrogate specimens, has long been a leading technology for revealing the shape and structure of the tiniest objects, from the cells which make up the bodies organs and microbes to individual building blocks or molecules which comprise cells, in often dramatic three-dimensional detail. But current EM techniques are limited in that they produce images only in grayscale, with colorization added later. In a paper published online November 3 in Cell Chemical Biology, researchers at University of California San Diego School of Medicine and Howard Hughes Medical Institute describe a new form of multicolor EM that allows for simultaneous visualization of multiple molecular species. "The ability to discern multiple specific molecules simultaneously adds a new dimension. It reveals details, actions and processes that aren't necessarily visible -- or even suspected -- in a more monochromatic view," said Mark H. Ellisman, PhD, professor in the Department of Neurosciences and director of the National Center for Microscopy and Imaging Research. Ellisman is a co-senior author of the study. Roger Tsien, PhD, professor of pharmacology, chemistry and biochemistry, was also co-senior author. Tsien, who passed away August 24, was co-winner of the 2008 Nobel Prize in chemistry for his work developing green fluorescent proteins as an imaging research tool. He was considered a leading light in microscopy and imaging research and was cited as one of the "world's most influential scientific minds" by Thomson Reuters earlier this year. Although there have been major improvements in multicolor and super-resolution fluorescence microscopy in recent years, comparable progress in EM has been more limited, achieved through automation and developments like the miniSOG protein, a new type of genetic tag visible under an EM microscope that was developed by Tsien, Ellisman and colleagues in 2011. To create multicolor EM images, first author and project scientist Stephen R. Adams, PhD, said researchers sequentially painted cellular structures such as proteins, membranes or whole cells with different "rare earth" metals, such as lanthanum, cerium and praseodymium in the form of precipitates. "A transmission electron microscope can distinguish each of these metals by electron energy-loss to give elemental maps of each that can be overlaid in color on the familiar monochrome electron micrograph," said Adams. "Each color highlights a different component of the cellular ultrastructure." Multicolor EM offers the possibility to differentiate detail not possible with standard EM, which uses gold particles to label structures but which appear in images as sometimes hard-to-distinguish black spots. It provides spatial resolution not possible with fluorescence microscopy. "This new method gives a more complete and easily detectable readout of the cellular components as colors," said Adams. "In theory, we should be able to add many more colors if we can develop more ways of precipitating additional lanthanides. The method is quite simple to do, uses easily made chemicals and requires detectors that are already present on many transmission electron microscopes so it is potentially readily transferable to other laboratories. Further research is needed to improve the chemistry and sensitivity of the method, but this work will hopefully inspire other groups to devise similar methods in this field."


But current EM techniques are limited in that they produce images only in grayscale, with colorization added later. In a paper published online November 3 in Cell Chemical Biology, researchers at University of California San Diego School of Medicine and Howard Hughes Medical Institute describe a new form of multicolor EM that allows for simultaneous visualization of multiple molecular species. "The ability to discern multiple specific molecules simultaneously adds a new dimension. It reveals details, actions and processes that aren't necessarily visible—or even suspected—in a more monochromatic view," said Mark H. Ellisman, PhD, professor in the Department of Neurosciences and director of the National Center for Microscopy and Imaging Research. Ellisman is a co-senior author of the study. Roger Tsien, PhD, professor of pharmacology, chemistry and biochemistry, was also co-senior author. Tsien, who passed away August 24, was co-winner of the 2008 Nobel Prize in chemistry for his work developing green fluorescent proteins as an imaging research tool. He was considered a leading light in microscopy and imaging research and was cited as one of the "world's most influential scientific minds" by Thomson Reuters earlier this year. Although there have been major improvements in multicolor and super-resolution fluorescence microscopy in recent years, comparable progress in EM has been more limited, achieved through automation and developments like the miniSOG protein, a new type of genetic tag visible under an EM microscope that was developed by Tsien, Ellisman and colleagues in 2011. To create multicolor EM images, first author and project scientist Stephen R. Adams, PhD, said researchers sequentially painted cellular structures such as proteins, membranes or whole cells with different "rare earth" metals, such as lanthanum, cerium and praseodymium in the form of precipitates. "A transmission electron microscope can distinguish each of these metals by electron energy-loss to give elemental maps of each that can be overlaid in color on the familiar monochrome electron micrograph," said Adams. "Each color highlights a different component of the cellular ultrastructure." Multicolor EM offers the possibility to differentiate detail not possible with standard EM, which uses gold particles to label structures but which appear in images as sometimes hard-to-distinguish black spots. It provides spatial resolution not possible with fluorescence microscopy. "This new method gives a more complete and easily detectable readout of the cellular components as colors," said Adams. "In theory, we should be able to add many more colors if we can develop more ways of precipitating additional lanthanides. The method is quite simple to do, uses easily made chemicals and requires detectors that are already present on many transmission electron microscopes so it is potentially readily transferable to other laboratories. Further research is needed to improve the chemistry and sensitivity of the method, but this work will hopefully inspire other groups to devise similar methods in this field." Explore further: Rejuvenating electron microscopy: Scientists modify plant protein to provide way to see previously unseen More information: "Multicolor electron microscopy for simultaneous visualization of multiple molecular species" Cell Chemical Biology, DOI: 10.1016/j.chembiol.2016.10.006


News Article | November 17, 2016
Site: www.eurekalert.org

Eight million people per year in the UK suffer from muscular diseases and injuries including muscular dystrophy, cerebral palsy, exercise-related injuries, rotator cuff tears, and age-related muscle loss (1). A new form of 3D imaging of muscles has allowed researchers to "see" inside muscle and trace long cables made up of a protein called collagen. Collagen cables are one culprit behind muscular diseases and injuries, so targeting them could provide treatments. That's according to new research from a team at UC San Diego and the Rehabilitation Institute of Chicago (RIC) led by Dr. Richard Lieber, currently Chief Scientific Officer at RIC and published in The Journal of Physiology. Muscular conditions, whether hereditary, exercise-induced, or due to normal aging, can result in stiff, dysfunctional muscles due to changes called fibrosis. Fibrosis is a roadblock to muscle recovery, and can result in muscle pain, weakness, limited range of motion, or require surgery to treat it. Researchers used a mouse model of skeletal muscle fibrosis to investigate the structure and function of collagen. They visualized collagen with conventional 2D and a newly developed method of 3D electron microscopy, mechanically measured muscle stiffness, and quantified the collagen producing cells. Dr. Allison Gillies, first author of the study, said: "The first time we looked at the 3D imaging results we were surprised--the collagen structures that we saw did not fit the textbook definition of muscle." Collagen had not been previously known to form long chains in muscle. Researchers had seen collagen outside muscle cells, but had not determined the level of organization that could only be visualized by 3D microscopy. When muscles become fibrotic, the number of cables and cells that produce collagen both increase. These collagen cables and the cells that produce collagen are thus two enticing targets for treating muscle disease and injury. Commenting on the study, senior author, Dr. Lieber said: "Reducing the amount of collagen cables or collagen producing cells in fibrotic muscle may improve muscle function and reduce pain, even obviating the need for corrective surgery." Rotator cuff tear- UK Census (http://www. ); Yamaguchi K, Ditsios K, Middleton WD et al 2006. The demographic and morphological features of rotator cuff disease. A comparison of asymptomatic and symptomatic shoulders. Journal of Bone & Joint Surgery (Am) 88(8):1699-704; and Tempelhof S, Rupp S, Seil R 1999. Age-related prevalence of rotator cuff tears in asymptomatic shoulders. Journal of Shoulder & Elbow Surgery 8(4):296-9. Sarcopenia- UK Census (http://www. ) and Patel HP, Syddall HE, Jameson K et al 2013. Prevalence of sarcopenia in community-dwelling older people in the UK using the European Working Group on Sarcopenia in Older People (EWGSOP) definition: findings from the Hertfordshire Cohort Study (HCS). Age & Ageing 42(3):378-84. Exercise related injuries- Nicholl JP, Coleman P, Williams BT 1991. Pilot study of the epidemiology of sports injuries and exercise-related morbidity. British Journal of Sports Medicine 25(1):61-6; and S Boyce and M Quigley 2004. Review of sports injuries presenting to an accident and emergency department. Emergency Medicine Journal 21(6):704-706. 3. The Journal of Physiology publishes advances in physiology that increase our understanding of how our bodies function in health and disease. http://jp. 4. The Physiological Society brings together over 3,500 scientists from over 60 countries. The Society promotes physiology with the public and parliament alike. It supports physiologists by organizing world-class conferences and offering grants for research and also publishes the latest developments in the field in its three leading scientific journals, The Journal of Physiology, Experimental Physiology and Physiological Reports. http://www. 5. The research was performed at UC San Diego and the Rehabilitation Institute of Chicago under Dr. Richard Lieber in collaboration with the National Center for Microscopy and Imaging Research led by Dr. Mark Ellisman. a. While the mouse model is genetically similar to human desminopathy, it differs from human muscle fibrosis because the mice do not show the variety of disease severity seen in humans. b. Three-dimensional reconstructions sampled a small volume of fibrotic muscle due to the technical limitations of this imaging method. There may be additional changes in muscle that the researchers have not seen yet. c. Collagen is one of many proteins in the extracellular matrix. The researchers studied it because it's the most abundant. There may be other proteins altered with fibrosis that may affect muscle stiffness and function.

Loading National Center for Microscopy and Imaging Research collaborators
Loading National Center for Microscopy and Imaging Research collaborators