Imaging  

Adopting new imaging technology into your business could give you an edge over your competitors in your field.  The equipment used in imaging systems is flexible enough to be helpful across a wide variety of industries, from automotive manufacturing to healthcare and retail distribution.

Imaging technology plays an increasingly important role in today's businesses. As a part of the digital age, imaging equipment allows for the rapid input of data. This data can then be used to boost security, safety, productivity and performance of the business.

Increased Security

Imaging technology also plays an important role in the security of businesses. In-house photographic imaging and printing systems allow visitors to be photographed and given a badge that displays the photo. This is entered into a computer system so that security personnel have an accurate count of how many people are in the building or on the premises. The security imaging technology can also be integrated with computer chips for use in employee access cards.

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• New Microscope Animates Life Processes at Subcellular Level 
• New Imaging Technique Detects Tumors Without Blood Tests or Biopsies 
• Implanted Chip Will Provide 3D Images from Inside the Heart 

Improved Automotive Safety

Today's cars and trucks feature high-tech, digital dashboards that are as easy to use as an iPad or the touchscreen of a smartphone. This is done through LED displays that are back-lit for easy viewing, especially when you are driving at night. Another automotive innovation in imaging is through digitally imaged defroster lines by companies such as Schilling Graphics. These grid lines on back windshields allow for the soldering between heat strips. The imaged lines allow drivers to see safely even while the defrosting system is in use.

Accurate Inventory

Imaging also allows for more accurate inventory techniques and procedures. Large retailers and other businesses that stock a vast amount of different supplies and materials benefit from the QR code technology and other imaging tools that allows for the rapid and accurate tracking of inventory. Businesses can even implement algorithms that allows for the automatic reordering of supplies when the inventory in stock drops below a specific level.

Enhanced Productivity

In businesses such as health care and medical care, imaging technology allows for a faster throughput of patients. Having an in-house digital X-ray machine for imaging allows doctors to perform an X-ray and have it read while the patient waits. Imaging technology in dental care allows for a patient's mouth to be scanned with a laser and the 3D images transferred to the software so that customized caps, crowns, bridges and dentures can be made.

Imaging technology delivers an excellent return on investment. The equipment used in imaging systems is flexible enough to be helpful across a wide variety of industries, from automotive manufacturing to healthcare and retail distribution. Bringing new imaging technology into your business could give you an edge over your competitors in the field.

By  Lizzie Weakley	Embed
About the Author: Lizzie Weakley is a freelance writer from Columbus, Ohio. She went to college at The Ohio State University where she studied communications. In her free time, she enjoys the outdoors and long walks in the park with her 3-year-old husky Snowball.

Imaging  

New imaging methods dramatically improve the spatial resolution provided by structured illumination microscopy, one of the best imaging techniques for seeing inside living cells.

Scientists can now watch dynamic biological processes with unprecedented clarity in living cells using new imaging techniques developed by researchers at the Howard Hughes Medical Institute's Janelia Research Campus. The new methods dramatically improve on the spatial resolution provided by structured illumination microscopy (SIM), one of the best imaging methods for seeing inside living cells.

The vibrant videos produced with the new technology show the movement and interactions of proteins as cells remodel their structural supports or reorganize their membranes to take up molecules from outside the cell. Janelia group leader Eric Betzig, postdoctoral fellow Dong Li and their colleagues have added the two new technologies — both variations on SIM — to the set of tools available for super-resolution imaging. Super-resolution optical microscopy produces images whose spatial resolution surpasses a theoretical limit imposed by the wavelength of light, offering extraordinary visual detail of structures inside cells. But until now, super-resolution methods have been impractical for use in imaging living cells.

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“These methods set a new standard for how far you can push the speed and non-invasiveness of super-resolution imaging,” Betzig says of the techniques his team described in a recent, issue of the journal Science. “This will bring super-resolution to live-cell imaging for real.”

In traditional SIM, the sample under the lens is observed while it is illuminated by a pattern of light (more like a bar code than the light from a lamp). Several different light patterns are applied, and the resulting moiré patterns are captured from several angles each time by a digital camera. Computer software then extracts the information in the moiré images and translates it into a three-dimensional, high-resolution reconstruction. The final reconstruction has twice the spatial resolution that can be obtained with traditional light microscopy.

Betzig was one of three scientists awarded the 2014 Nobel Prize in Chemistry for the development of super-resolved fluorescence microscopy. He says SIM has not received as much attention as other super-resolution methods largely because those other methods offer more dramatic gains in spatial resolution. But he notes that SIM has always offered two advantages over alternative super-resolution methods, including photoactivated localization microscopy (PALM), which he developed in 2006 with Janelia colleague Harald Hess.

“I fell in love with SIM because of its speed and the fact that it took so much less light than the other methods,” Betzig says.

Betzig began working with SIM shortly after the death in 2011 of one of its pioneers, Mats Gustafsson, who was a group leader at Janelia. Betzig was already convinced that SIM had the potential to generate significant insights into the inner workings of cells, and he suspected that improving the technique's spatial resolution would go a long way toward increasing its use by biologists.

Betzig's new method was simple. He says: “Don't turn on all of the molecules. There's no need to do that.”

Instead, the new method, called patterned photoactivation non-linear SIM, begins by switching on just a subset of fluorescent labels in a sample with a pattern of light. “The patterning of that gives you some high resolution information already,” he explains. A new pattern of light is used to deactivate molecules, and additional information is read out of their deactivation. The combined effect of those patterns leads to final images with 62-nanometer resolution—better than standard SIM and a three-fold improvement over the limits imposed by the wavelength of light.

“We can do it and we can do it fast,” he says. That's important, he says, because for imaging dynamic processes, an increase in spatial resolution is meaningless without a corresponding increase in speed. “If something in the cell is moving at a micron a second and I have one micron resolution, I can take that image in a second. But if I have 1/10-micron resolution, I have to take the data in a tenth of a second, or else it will smear out,” he explains.

"Most of the magic is in the software, not the hardware."

Patterned photoactivation non-linear SIM captures the 25 images that go into a final reconstruction in about one-third of a second. Because it does so efficiently, using low intensity light and gleaning information from every photon emitted from a sample's fluorescent labels, labels are preserved so that the microscope can image longer, letting scientists watch more action unfold.

The team used patterned photoactivation non-linear SIM to produce videos showing structural proteins break down and reassemble themselves as cells move and change shape, as well as the dynamics of tiny pits on cell surfaces called caveolae.

Betzig's team also reports in the Science paper that they can boost the spatial resolution of SIM to 84 nanometers by imaging with a commercially available microscope objective with an ultra-high numerical aperture. The aperture restricts light exposure to a very small fraction of a sample, limiting damage to cells and fluorescent molecules, and the method can be used to image multiple colors at the same time, so scientists can simultaneously track several different proteins.

Using the high numerical aperture approach, Betzig's team was able to watch the movements and interactions of several structural proteins during the formation of focal adhesions, physical links between the interior and exterior of a cell. They also followed the growth and internalization of clathrin-coated pits, structures that facilitate the intake of molecules from outside of the cell. Their quantitative analysis answered several questions about the pits' distribution and the relationship between pits' size and lifespan that could not be addressed with previous imaging methods.

Finally, by combining the high numerical-aperture approach with patterned photoactivatable non-linear SIM, Betzig and his colleagues could follow two proteins at a time with higher resolution than the high numerical aperture approach offered on its own.

Betzig's team is continuing to develop their SIM technologies, and say further improvements are likely. They are also eager to work with biologists to continue to explore potential applications and refine their techniques' usability.

For now, scientists who want to experiment with the new SIM methods can arrange to do so through Janelia's Advanced Imaging Center, which provides access to cutting-edge microscopy technology at no cost. Eventually, Betzig says, it should be fairly straightforward to make the SIM technologies accessible and affordable to other labs. “Most of the magic is in the software, not the hardware,” he says.

SOURCE  Howard Hughes Medical Institute

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 Imaging  

Tumors are often surrounded and invaded by bone marrow-derived cells. Imaging the infiltration of such immune cells into tumors may therefore be an attractive means of detecting tumors or of tracking the response to anticancer therapy. A new imaging technique may now make this possible.

Using a new approach has allowed real-time imaging of the immune system’s response to the presence of tumors—without the need for blood draws or invasive biopsies. The method offers a potential breakthrough both in diagnostics and in the ability to monitor efficacy of cancer therapies.

The method, developed in the lab of Whitehead Institute Member Hidde Ploegh and reported in the Proceedings of the National Academy of Sciences (PNAS), harnesses the imaging power of positron emission tomography (PET), which is normally used to monitor cancer metabolism, to identify areas of immune cell activity associated with inflammation or tumor development.

“Every experimental immunologist wants to monitor an ongoing immune response, but what are the options?” Ploegh asks rhetorically. “One can look at blood, but blood is a vehicle of transport for immune cells and is not where immune responses occur. Surgical biopsies are invasive and non-random, so, for example, a fine-needle aspirate of a tumor could miss a significant feature of that condition.”

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In search of a better monitoring approach Ploegh leveraged two research tools that have become staples in his lab in recent years. The first exploits so-called single-domain antibodies known as VHHs, derived from the heavy chain-only antibodies made by the immune systems of animals in the camelid family. Ploegh’s lab immunizes alpacas to generate VHHs specific to immune cells of interest. The second tool, known as sortagging, labels the VHHs in site-specific fashion to enable the tracking of the VHHs and their targets in a living animal.

"We’re very excited about this because it’s a powerful approach to pick up inflammation in and around the tumor."

Knowing that the tissue around tumors often contains immune cells such as neutrophils and macrophages, Ploegh and his lab members hypothesized that appropriately labeled VHHs might allow them to pinpoint tumor locations by finding the tumor-associated immune cells. Ploegh notes that VHHs’ extremely small size—approximately one-tenth that of conventional antibodies—are likely responsible for their superior tissue penetration and thus makes them particularly well suited for such use.

For the current research, the lab generated VHHs that recognize mouse immune cells, then labeled these VHHs with radioisotopes, and injected them into tumor-bearing mice. Subsequent PET imaging detected the location of immune cells around the tumor quickly and accurately.

“We were able to image tumors as small as one millimeter in size and within just a few days of their starting to grow,” says Mohammad Rashidian, a postdoctoral researcher in Ploegh’s lab and first author of the PNAS paper. “We’re very excited about this because it’s a powerful approach to pick up inflammation in and around the tumor.”

Rashidian and Ploegh believe that with further refinement, the method could be used to monitor response to—and perhaps modify—cancer immunotherapy, which, though quite promising, has thus far met with great success in some cases, but has failed in others.


SOURCE  Whitehead Institute

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 Imaging  

Researchers have captured the first X-ray portraits of living bacteria, detecting signals from features as small as 4 nanometers, or 4 billionths of a meter.  This milestone sets the stage for X-ray explorations of the molecular machinery at work in viral infections, cell division, photosynthesis and other processes that are important to biology, human health and our environment.

Researchers working at the Department of Energy’s SLAC National Accelerator Laboratory and Uppsala University have taken the first X-ray portraits of living bacteria.

This milestone, reported in the journal Nature Communications, is a first step toward possible X-ray explorations of the molecular machinery at work in viral infections, cell division, photosynthesis and other processes that are important to biology, human health and our environment. The experiment took place at SLAC’s Linac Coherent Light Source (LCLS) X-ray laser, a DOE Office of Science User Facility.

“We have developed a unique way to rapidly explore, sort and analyze samples, with the possibility of reaching higher resolutions than other study methods,” said Janos Hajdu, a professor of biophysics at Uppsala University in Sweden, which led the research. “This could eventually be a complete game-changer.”

The experiment focused on cyanobacteria, or blue-green algae, an abundant form of bacteria that transformed Earth’s atmosphere 2.5 billion years ago by releasing breathable oxygen, making possible new forms of life that are dominant today. Cyanobacteria play a key role in the planet’s oxygen, carbon and nitrogen cycles.

Researchers sprayed living cyanobacteria in a thin stream of humid gas through a gun-like device. The cyanobacteria were alive and intact when they flew into the ultrabright, rapid-fire LCLS X-ray pulses, producing diffraction patterns recorded by detectors.

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The diffraction patterns preserved details of the living cyanobacteria that were compiled to reconstruct 2D images. Researchers said it should be possible to produce 3D images of some samples using the same technique.

The technique works with live bacteria and requires no special treatment of the samples before imaging. Other high-resolution imaging methods may require special dyes to increase the contrast in images, or work only on dead or frozen samples.

The technique can capture about 100 images per second, amassing many millions of high-resolution X-ray images in a single day. This speed allows sorting and analysis of the inner structure and activity of biological particles on a massive scale, which could be arranged to show the chronological steps of a range of cellular activities.

In this way, the technique merges biology and big data, said Tomas Ekeberg, a biophysicist at Uppsala University. “You can study the full cycle of cellular processes, with each X-ray pulse providing a snapshot of the process you want to study,” he said.

"We have developed a unique way to rapidly explore, sort and analyze samples, with the possibility of reaching higher resolutions than other study methods. This could eventually be a complete game-changer."

Hajdu added, “One can start to analyze differences and similarities between groups of cellular structures and show how these structures interact: What is in the cell? How is it organized? Who is talking to whom?”

While optical microscopes and X-ray tomography can also produce high-resolution 3D images of living cells, LCLS, researchers say, could eventually achieve much better resolution – down to fractions of a nanometer, or billionths of a meter, where molecules and perhaps even atoms can be resolved.

LCLS is working with researchers to improve the technique and upgrade some instruments and the focus of its X-rays as part of the LCLS Single-Particle Imaging initiative, formally launched at SLAC in October in cooperation with the international scientific community. The initiative is working toward atomic-scale imaging for many types of biological samples, including living cells, by identifying and addressing technical challenges at LCLS.




SOURCE  SLAC National Accelerator Laboratory

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 Medicine  

In hospitals and medical facilities of the not-too-distant future, medical imaging will be way beyond the realm of the traditional X-ray.  Already the latest generation of CT scanners is providing an unparalleled view inside our bodies, aiding diagnosis and treatment.

The ability to explore inside the human body non-invasively is a modern day medical miracle. Computed Tomography, or CT scans, as they’re more commonly known, use radiation to see inside a patient’s body.

CT scanners are often the first imaging technology many patients encounter when doctors suspect serious disease or injury. The machines use a narrow beam of X-rays processed by a computer to create slices of the body and assemble them into detailed 3D images.
Using computed tomography, doctors track organ function in real-time and do things science fiction writers in decades past could have only dreamed about.

Circle of Willis 

The set of high-resolution CT scan images in this post come from GE Healthcare’s Revolution CT, which was unveiled in a hospital setting for the first time in September of last year at West Kendall Baptist Hospital in Florida.

GE's Revolution CT delivers uncompromised image quality and clinical capabilities through the convergence of coverage, spatial and temporal resolution.

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The scanner not only produces high-quality shots of the organs, but delivers lower doses of radiation to achieve them, putting patients at greater ease and less risk.

“According to our physicians, patient feedback about their experience with the Revolution CT has been uniformly positive,” said Javier Hernández-Lichtl, CEO of West Kendall Baptist, in a GE company report. “The advanced design definitely makes for a less intimidating, more comfortable patient experience, while yielding amazingly accurate and detailed images.”

Physicians have already collected images of major blood vessels, organs, bones, and tissues. They can watch blood pump through patients’ veins and see recently installed hardware, like coronary stents, keep arteries clear as oxygen-rich blood enters the heart.

In addition, the speed of the new technology allows providers to gather information about function as well as anatomy, enabling a comprehensive stroke assessment of the brain in a single exam.

“A core component of our strategy at GE Healthcare,” said Jeff Immelt, GE chairman and CEO, “is to partner with customers to understand their clinical and operational needs, and in turn develop next-generation technology that deliver the necessary outcomes.”

SOURCE  Business Wire, GE Healthcare

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 Main Label  

NIH researchers have found that the biological machinery that builds DNA can insert molecules into the DNA strand that are damaged as a result of environmental exposures. These damaged molecules trigger cell death that produces some human diseases, according to the researchers.

By using a new imaging technique, National Institutes of Health (NIH) scientists have found that the biological machinery that builds DNA can insert molecules into the DNA strand that are damaged as a result of environmental exposures. These damaged molecules trigger cell death that produces some human diseases, according to the researchers.

The work, appearing in the journal Nature, provides a possible explanation for how one type of DNA damage may lead to cancer, diabetes, hypertension, cardiovascular and lung disease, and Alzheimer’s disease.

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Time-lapse crystallography was used by National Institute of Environmental Health Sciences (NIEHS) researchers to determine that DNA polymerase, the enzyme responsible for assembling the nucleotides or building blocks of DNA, incorporates nucleotides with a specific kind of damage into the DNA strand. Time-lapse crystallography is a technique that takes snapshots of biochemical reactions occurring in cells.

Samuel Wilson, M.D., senior NIEHS researcher on the team, explained that the damage is caused by oxidative stress, or the generation of free oxygen molecules, in response to environmental factors, such as ultraviolet exposure, diet, and chemical compounds in paints, plastics, and other consumer products. He said scientists suspected that the DNA polymerase was inserting nucleotides that were damaged by carrying an additional oxygen atom.

“When one of these oxidized nucleotides is placed into the DNA strand, it can’t pair with the opposing nucleotide as usual, which leaves a gap in the DNA,” Wilson said. “Until this paper, no one had actually seen how the polymerase did it or understood the downstream implications.”

"Until this paper, no one had actually seen how the polymerase did it or understood the downstream implications."

Wilson and his colleagues saw the process in real time, by forming crystal complexes made of DNA, polymerase, and oxidized nucleotides, and capturing snapshots at different time points through time-lapse crystallography. The procedure not only uncovered the stages of nucleotide insertion, but indicated that the new DNA stopped the DNA repair machinery from sealing the gap. This fissure in the DNA prevented further DNA repair and replication, or caused an immediate double-strand break.

“The damaged nucleotide site is akin to a missing plank in a train track,” Wilson said. “When the engine hits it, the train jumps the track, and all of the box cars collide.”

Large numbers of these pileups and double-strand breaks are lethal to the cell, serving as a jumping off point for the development of disease. However, it can be a good thing if you are a researcher trying to destroy a cancer cell.

“One of the characteristics of cancer cells is that they tend to have more oxidative stress than normal cells,” said Bret Freudenthal, Ph.D., lead author of the paper and postdoctoral fellow in Wilson’s group. “Cancer cells address the issue by using an enzyme that removes oxidized nucleotides that otherwise would be inserted into the genome by DNA polymerases. Research performed by other groups determined if you inhibit this enzyme, you can preferentially kill cancer cells.”

Wilson and Freudenthal stressed that the quantities of oxidized nucleotides in the nucleotide pool are usually under tight control, but if they accumulate and start to outnumber undamaged nucleotides, the DNA polymerase adds more of them to the strand.

Molecules that inhibit oxidation, known as antioxidants, reduce the level of oxidized nucleotides, and may help prevent some diseases.



SOURCE  NIH

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 Health  

Modern dentistry is almost entirely focused on the use of computers and imaging technology to help people maintain brighter smiles. Here are some of the cutting-edge technologies you will find in a modern dentist's office.

Dentistry has come a long way since its medieval days of brushing with wine and wooden dentures. Modern dentistry is almost entirely focused on the use of computers and imaging technology to help people maintain brighter smiles. Modern dentists can insert dental implants and prevent the rotting of teeth in a number of different ways. These dentists and orthodontists can do almost everything to improve teeth except aid in the re-growing of the tooth itself.

3D Imaging Technology

Dental technology is slowly moving its focus into complex 3D imaging to replace the use of X-rays. This 3D modeling software is able to completely reconstruct a 3D model of the patient's mouth. The doctor can manipulate the model in 3D space to look for weakness or signs of decay within the teeth.

Like sneak previews? Dentists and orthodontists show you how your mouth will look once a procedure is finished. Are you a fan of movies? Powerful imaging technology can create a movie of how your teeth have changed over time and will continue to change. This can be especially encouraging while enduring the torture of braces.

The technology to create realistic models of teeth with relative ease already exists in many modern dentist offices.

Invisible Braces

Speaking of terrible braces, the “railroad track” style of braces are on their way out of the medical field. According to orthodontists at Able Dental Group, Invisalign brand braces are taking the world of teeth by storm. Every two weeks, your dentist replaces the invisible braces with custom fit aligners that can more than handle overbite, underbite, gaps, and overcrowded teeth. Forget cut lips, broken brackets and brace wax, there’s a new sleek product in town.

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CAD/CAM Dentistry

CAD/CAM dentistry is similar to 3D imaging, but it's used mostly for the creation of dental restorations. Dentists can create more accurate recreations of bridges and crowns using this technology. A single cobalt disc can contain enough dental crowns to cap a patient's entire mouth. CAD/CAM technology can also use 3D models to recreate a patient's mouth from old dental records and give them a set of dentures that is very close to how their teeth originally looked.

Diode Lasers

Many dentist offices are now adopting diode lasers over more traditional hygiene tools. These lasers are used for cosmetic dentistry and also some kinds of surgery. Many diode lasers are wireless and don't require the use of pedals. Dentists can carefully perform tissue procedures with these lasers that result in more comfort for the patient and less of a need for anesthetic. Diode lasers cost only a few thousand dollars, so expect to see them in more dental offices in the near future.

By Hannah Whittenly

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 Imaging  

Phase contrast X-ray imaging has enabled researchers to perform mammographic imaging that allows greater precision in the assessment of breast cancer and its precursors. The technique could improve biopsy diagnostics and follow-up.

Researchers have succeeded in advancing an emerging imaging technique for breast investigations: the X-ray phase-contrast mammography. The new developments enable distinguishing between the different types of microcalcifications observed in breast tissue and help assigning them to malignant lesions. The study has just been published in Nature Communications.

One of the advantages of the phase contrast technique is its ability to provide images of high contrast. In the future, this technique can aid physicians to determine in a non-invasive way where pre-malignant and malignant breast lesions are most likely located.

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One goal of breast cancer screening is to detect groups of microcalcifications in the breast, because these may be associated with early stages of breast cancer. Microcalcifications often occur in connection with cancer cell death. Mammographic screening does not allow definite conclusions regarding the underlining conditions that cause these calcifications. Only tissue biopsies that are examined under the microscope by pathologists can determine which lesions have caused the calcareous deposits.

"My observation could be very interesting for breast cancer diagnosis, since it could distinguish between the different types of microcalcifications."

At the ETH Zurich, the Paul Scherrer Institute (PSI), the use of phase contrast for medical X-ray imaging has been investigated for several years. X-ray radiation as used in conventional mammography was long considered not suitable for phase contrast procedures because of its incoherence and mixture of multiple wavelengths.

“The fact that we have now managed to use these X-ray sources for the phase contrast method in order to develop a new and improved imaging method is a considerable step towards application in daily clinical practice,” says Marco Stampanoni, Professor at the Institute for Biomedical Engineering at ETH Zurich and Head of the X-ray Tomography Group at the PSI.

In X-ray phase contrast, the extent in which tissue absorbs X-rays is not the only quantity that is being measured but also how tissue deflects radiation laterally (refraction) and consequently how it influences the sequence of oscillation peaks and valleys of X-ray waves – the so-called phase.

Depending on the tissue type, the overall scattering also varies. To be able to measure the phase shift, researchers use three very fine grids. The first one is located directly at the source. It ensures that the object is illuminated with the required coherence. Another grid is placed behind the object and generates an interference signal that is analysed by a third grid downstream. Using suitable algorithms, the researchers calculate the absorption, phase and scattering properties of the object from the interference signal. This information can be used to generate sharp and high-contrast images that show very detailed soft tissue properties.

A discovery by Zhentian Wang, PostDoc in Prof. Stampanoni’s team, initiated this development: “During my trials with the phase contrast method, I noticed that there are microcalcifications with different absorption and scattering signals. That indicated that the new method might identify different types of calcifications,” he says. Wang subsequently reviewed through medical literature and found studies that showed that a certain type of calcification is more frequently associated with breast cancer precursors. “I was persuaded that my observation could be very interesting for breast cancer diagnosis, since it could distinguish between the different types of microcalcifications”, says the researcher.

To date, the researchers have worked with a prototype. They examined breast tissue samples, but no patients have been involved yet. “One of our next aims will be to develop a device for clinical use,” says Marco Stampanoni.


SOURCE  ETH Zurich

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 Imaging  

A research team has developed a general method to image a broad spectrum of small biomolecules, such as small molecular drugs and nucleic acids, amino acids, lipids for determining where they are localized and how they function inside cells.

Researchers at Columbia University have made a significant step toward visualizing small biomolecules inside living biological systems with minimum disturbance, a longstanding goal in the scientific community. In a study published in Nature Methods, Assistant Professor of Chemistry Wei Min's research team has developed a general method to image a broad spectrum of small biomolecules, such as small molecular drugs and nucleic acids, amino acids, lipids for determining where they are localized and how they function inside cells.

When studying biological functions of a molecule in complex and mysterious cells, researchers typically label the molecules of interest with fluorophores, a kind of molecule that glows when illuminated. Using a fluorescence microscope, common in research labs, the fluorophore-tagged molecules can be located and tracked with high precision. The invention of green fluorescent protein (GFP), in 1994, compatible with imaging inside live cells and animals, has since made fluorescence microscopy even more popular.

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However, when it comes to small biomolecules, fluorophore tagging is problematic, because the fluorophores are almost always larger or comparable in size to the small molecules of interest. As a result, they often disturb the normal functions of these small molecules with crucial biological roles.

To address this problem, Min and his team departed from the conventional paradigm of fluorescence imaging of fluorophores, and pursued a novel combination of physics and chemistry. Specifically, they coupled an emerging laser-based technique called stimulated Raman scattering (SRS) microscopy with a small but highly vibrant alkyne tag (that is, C=C, carbon-carbon triple bond), a chemical bond that, when it stretches, produces a strong Raman scattering signal at a unique "frequency" (different from natural molecules inside cells).

This new technique, labeling the small molecules with this tiny alkyne tag, avoids perturbation that occurs with large fluorescent tags, while obtaining high detection specificity and sensitivity by SRS imaging. By tuning the laser colors to the alkyne frequency and quickly scanning the focused laser beam across the sample, point-by-point, SRS microscopy can pick up the unique stretching motion of the C=C bond carried by the small molecules and produce a three-dimensional map of the molecules inside living cells and animals.

In this way, Min's team demonstrated tracking alkyne-bearing drugs in mouse tissues and visualizing de novo synthesis of DNA, RNA, proteins, phospholipids and triglycerides through metabolic incorporation of alkyne-tagged small precursors in living cells.

"The major advantages of our technique lie in the superb sensitivity, specificity and biocompatibility with dynamics of live cells and animals for small molecule imaging," says the lead author Lu Wei, a Ph.D. candidate in chemistry.

Next, Min's team will apply this new technique to biomedical questions, such as detecting tumor cells and probing drug pharmacokinetics in animal models. They are also creating other alkyne-labeled biologically active molecules for more versatile imaging applications.

"Our new technique will open up numerous otherwise difficult studies on small biomolecules in live cells and animals," says Min. "In addition to basic research, our technique could also contribute greatly to translational applications. I believe SRS imaging of alkyne tags could do for small biomolecules what fluorescence imaging of fluorophores such as GFP has done for larger species."



SOURCE  Columbia University via EurekAlert

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 Imaging  

A new 3D imaging technique for live cells uses a conventional microscope to capture image slices throughout the depth of the cell, then computationally renders them into one three-dimensional image. The technique uses no dyes or chemicals, allowing researchers to observe cells in their natural state.

Living cells are ready for their close-ups, thanks to a new imaging technique that needs no dyes or other chemicals, yet renders high-resolution, three-dimensional, quantitative imagery of cells and their internal structures – all with conventional microscopes and white light.

Called white-light diffraction tomography (WDT), the imaging technique opens a window into the life of a cell without disturbing it and could allow cellular biologists unprecedented insight into cellular processes, drug effects and stem cell differentiation.

The team of University of Illinois researchers, led by electrical and computer engineering and bioengineering professor Gabriel Popescu, published their results in the journal Nature Photonics.

“One main focus of imaging cells is trying to understand how they function, or how they respond to treatments, for example, during cancer therapies,” Popescu said. “If you need to add dyes or contrast agents to study them, this preparation affects the cells’ function itself. It interferes with your study. With our technique, we can see processes as they happen and we don’t obstruct their normal behavior.”

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Because it uses white light, WDT can observe cells in their natural state without exposing them to chemicals, ultraviolet radiation, or mechanical forces – the three main methods used in other microscopy techniques. White light also contains a broad spectrum of wavelengths, thus bypassing the interference issues inherent in laser light – speckles, for example.

The 3D images are a composite of many cross-sectional images, much like an MRI or CT image. The microscope shifts its focus through the depth of the cell, capturing images of various focus planes. Then the computer uses the theoretical model and compiles the images into a coherent three-dimensional rendering.

The greatest potential of WDT, according to the researchers, is the ability to study cells in three dimensions over time. Since the cells are not altered, they can be imaged repeatedly, allowing researchers a glimpse into the dynamics of a cell as it goes about its life – or as it is treated with a new drug.

“As a cell grows we can see the change in all three dimensions,” said Taewoo Kim, a graduate student and first author of the paper. “We can see the dynamics of the cell in 3-D, which hasn’t been done in a quantitative manner. For example, we could see, in the span of a minute or over a cell’s lifetime, how it grows and how the things in the cell move around.”

“With this imaging we can tell at what scale things within the cell are transported randomly and at what scale processes are actually organized and deterministic,” Popescu said. “At first glance, the dynamics looks pretty messy, but then you look at it – we stare at movies for hours and hours – and you realize it all makes sense. Everything is organized perfectly at certain scales. That’s what makes a cell alive. Randomness is just nature’s way to try new things.”

WDT uses a component that adds onto a conventional phase contrast microscope, a common piece of equipment in biology labs, without altering the microscope itself. The researchers used conventional microscopes with the intention of making these new optics principles easily accessible for biologists. The researchers hope that this will allow rapid large-scale adoption of WDT, and Popescu founded a startup company, Phi Optics, to help achieve that goal.

In addition to biological applications, the WDT technique has implications in the broader field of optics as the researchers pushed the boundaries of physics by applying scattering theory to imaging optics.

“The physics behind this technique is another thing we were fascinated about,” Kim said. “Light propagation in general is studied with approximations, but we’re using almost no approximation. In a very condensed form, we can perfectly show how the light changes as it passes through the cell.”

“We started on this problem two years ago, trying to formulate mathematically the sectioning effect observed in spatial light interference light microscopy (SLIM),” said Renjie Zhou, a graduate student and co-first author of the paper. “We came up with equations which eventually described WDT. The final equation is beautiful and the theory opens opportunities for solving other optics problems in a new theoretical language.”

Next, the researchers hope to pursue cross-disciplinary collaborations to explore applications of WDT in biology as well as expansions of the imaging optics demonstrated in WDT. For example, they are using WDT to watch stem cells as they differentiate in hopes of better understanding how they turn into different cell types. Since stem cells are so sensitive, only a chemical-free, non-invasive, white-light technique such as WDT could be used to study them without adverse effects.



SOURCE  University of Illinois at Urbana-Champaign

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 Imaging  

RealView Imaging has developed an interactive 3D holographic system, initially for medical imaging applications that is remarkably similar to that used by Tony Stark in the Iron Man movies.

The world's first 3D holographic display and interface system, initially for medical imaging applications has been unveiled by RealView Imaging.

The system looks to be close to the science fiction version used by Tony Stark in the Iron Man movies.

The company has managed to invent and develop a system that can receive volumetric stream such as 3D Ultrasound or any other static or stream of volumes, convert it in real-time to interference patterns and project true holograms using these interference patterns.

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In traditional holography an interference pattern is created by splitting a coherent beam of light projecting one part onto an object and the other part, called a reference beam, on to a sensitive film in a location such that it will absorb at least some of the light scattered from the object. Later the film is developed and when lighted with a similar reference beam it reconstructs the object in free space as a hologram.

The holograms that are projected are with high resolution and quality, they are with full color and can be viewed from a very wide angle. Moreover, the reconstructed holograms are within the user’s touching reach and remain in the same coordinates, independent of the viewer’s location.

This proprietary capability enables precise and direct interaction with and within the images by literally touching the holograms and manipulating them in real-time. As these are true holograms there is a very high level of realism thus the viewer can intuitively comprehend even unique and complex 3D structures/pathologies and additionally the interaction with the images is as intuitive as grabbing an apple or painting a statue as the image is optically real and within touching reach..



SOURCE  RealView Imaging

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