Visualizing Science: Microscopic Images from UNC Charlotte - Digital Exhibit

The J. Murrey Atkins Library was pleased to host an exhibit and competition, Visualizing Science: Microscopic Images from UNC Charlotte, to highlight the important role of the University of North Carolina at Charlotte in scientific advancement. All faculty, staff, students, and alumni were invited to submit research images or "scientific art" produced with any type of microscope. The print images were displayed on the first floor of the Atkins Library from November 11 through December 9, 2015

Thumbnail images of each submission, along with the accompanying explanation of the research and how the image was produced, are provided on this webpage. A picture description is set below each image to be translated by a screen reader for the visually-impaired. All images are organized by participant department or program with quick links for ease of navigation.




Bioinformatics and Genomics

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Oryza

Green oval granule of rice covered with fine hair like filaments, which is emerging from an elongated leaf sheath. A single small white flower bud is positioned at the tip of the rice granule.

Tanner Deal, Nowlan Freese, Ph.D. Department of Bioinformatics and Genomics
This image is of a ripening granule of Oryza sativa (rice), one of the most important grain crops, accounting for 20% of the world’s total calories. This particular variety is called Agami, and is being studied for its remarkable ability to grow in soil with high levels of salt. Of the various cereal grains, rice is the most sensitive to salt, with yields decreased by up to 50% when grown in salty soils. This has a significant impact on the ability to grow rice, as 33% of the world’s arable lands are affected by excess salt. This variety of rice is being analyzed using genomic sequencing technologies along with observations of its growth and development in order to discover why it is able to grow so well in salty soil. Image taken on a Zeiss Axio Observer top-illuminated microscope at 10X magnification.



Biological Sciences

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Starvation by sea (to shining sea)

Sea urchin larva, a shuttlecock-like organism, with six slender larval arms projecting from the base and arranged in radial symmetry. This larva has similar features as sand dollar larvae.

Tyler J. Carrier Department of Biological Sciences
Many larvae of marine invertebrates actively feed for growth towards a developmental threshold where they transition to the ocean floor and become juveniles. This transition is often delayed due to the scarcity of food. Larvae facing an extensive period of starvation resort to inherited energetic reserves. Some larvae die soon after this buffer is depleted while others alter basic functions to extend life by entering a hibernation-like state where total energy expenditure is minimized. Larvae of sea urchins are best known for this phenomenon by elongating their arms and reabsorb stomach tissues. Moreover, they minimize metabolism activity, become hypersensitive to the environment, ramp up their immune system, and slow the 'aging' process. As a means to study starvation biology of sea urchin larvae and why this novelty was selected for as a 'default', I diet-restricted larvae of the green sea urchin Strongylocentrotus droebachiensis and examine the expression of certain molecular pathways as well as the symbiotic microbial community. This microphotograph was imaged at the MDI Biological Laboratory (Bar Harbor, ME) with assistance from Dr. James Coffman using an Olympus FVX10 at 100x.



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In the beginning...

Circular single cell sea urchin egg in the process of dividing into two cells and starting life as an embryo.

Tyler J. Carrier Department of Biological Sciences
For you and I as well as many other animals, life starts as a single cell that is activated to begin dividing at fertilization. At simplest form, as seen here, one cannot differentiate whether an embryo is that of a sea anemone, sea urchin, or human. Developmental biologists study how eggs mature into complex multi-cellular organisms, and how and why a developmental pattern differs between various animals, even though indistinguishable at simplest form, the early embryo. Throughout history biologist have noticed that normal development, or embryogenesis, and regeneration use many of the same functions. For more than a century echinoderm – for example, sea urchins and sea stars – were the primary model for developmental and regenerative biology. For my research I use sea urchins and their close relative, the sea star, to study the link between development and regeneration to improve human health. More specifically, I study the proteins and their complex interactions during regeneration, and where and when those proteins are utilized during development. This image is a 2-cell embryo of the purple sea urchin Strongylocentrotus purpuratus, and was taken in June 2015 at Brown University using a compound microscope at 400x.



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The Wonderful World of Yeast!

Black background with sixteen budding yeast cells in rows of four and columns of four. Alternating cells are overlaid with one gray circle sitting on the other, similar in form to a snowman. Splashes of bright red, pink, blue, aqua, yellow, purple, and orange colors appear throughout, including forming ghost-like images of the yeast in between the actual budding cells.

Richard J. Chi, Ph.D. Department of Cell Biology, Yale School of Medicine Department of Biological Sciences, University of North Carolina at Charlotte
Saccharomyces Cerevisiae also known as Baker’s/Brewer’s Yeast, were imaged using GE’s DeltaVision elite system which is a wide-field epi-fluorescence microscope that acquires images using an exclusive deconvolution algorithm. The living cells expressed three different fluorescent organelle markers; Autophagasomes (Green), Mitochondria (Red), Vacuoles (Blue), top row. Images were taken using a 100X objective equipped with differential-interference-contrast (DIC). Each color was taken in serial sections, and then recombined to form a 3-dimensional composite image. Subsequent series were pseudo-colored and DIC micrographs were overlaid using Adobe Photoshop. Despite the beautiful array of colors, this particular yeast is very unhappy. The genes that encode two very important proteins are missing. This results in enlarged autophagasomes, aberrant mitochondria and fragmented vacuoles. Similar mutations or defects in human versions of these proteins can lead to diseases such as Parkinson’s and Alzheimer’s. Using Yeast and fluorescent microscopy, researchers have identified important genes that cause many human diseases.



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And we'll all float on

Sand dollar larva, a shuttlecock-like organism, with eight slender larval arms projecting from the base and arranged in radial symmetry. This larva has similar features as sea urchin larvae.

Tyler J. Carrier Department of Biological Sciences
Many animals in the ocean, whether on the sea floor (benthic) or in the water column (pelagic), are invertebrates. A common reproductive approach used by benthic invertebrates is broadcast spawning, where adults release eggs or sperm into the water column to be fertilized and begin developing into larvae. These larvae serve as vectors to expand a species range, and to seed the next generation. But, how long do larvae remain larvae? For some species, larvae remain pelagic for a few hours or days while others endure a journey upwards of a year. Larvae that remain planktonic for weeks to months may be transported 10s to 100s or even 1000s of kilometers by ocean currents. In fact, larvae of the green sea urchin found in the Gulf of Maine are frequently transported across the North Atlantic Ocean to the Celtic Sea (bordering France, Ireland, and the United Kingdom). This extent of transport is common for species in the deep-sea, as they locate very sparse habitats that harbor a diversity of life. This microphotograph of a sand dollar larva was imaged at the Shannon Point Marine Center (Anacortes, WA) using an Olympus compound microscope at 100x.



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Oh the places you'll go

Sea star larva with a recently formed gut (by folding back onto itself to form an elongated cavity), that is shaped like a spherical cylinder with single cells that can be distinguished for single-cell migrations.

Tyler J. Carrier Department of Biological Sciences
Our most common recollection of migratory animals is monarch butterfly and birds, which annually relocate from New England to the Southeast. In the sea, lobsters, shrimp, and sea urchins are notorious for migrating from deep, cold offshore waters to shallower coastal waters in the late winter to early spring. But migration occurs at many other levels, including by single cells or populations of cells – as seen here in a sea star larva. During development from a single cell to a mature adult, cells may translocate from one sector of the animal to another, in which they may perform the same or a different set of tasks. In the image on the right, you can see single cells; and on some, you can also see string-like extension. These, termed pseudopodia, act as anchors so the cell may drag itself along a path to another location. This ability (to form pseudopodia) occurs throughout the animal kingdom – from single cell amoeba to humans. Image on the left and right are a late-stage gastrula embryo of the bat star Patiria miniata taken in June 2015 at Brown University using a compound microscope at 100x and 320x, respectively.



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Beautiful Cancer Death

Black background with a cluster of round to ovoid cells and the periphery of the cells is colored green. The interior of the cells contains blue stippling or spots that is DNA and red round to horseshoe shapes that represent a virus infection.

Eric Hastie, Ph.D. (2014) Department of Biological Sciences, Grdzelishvili Lab
Oncolytic virotherapy, the use of viruses to kill cancer, is an emerging therapeutic option for many cancer types. Here pancreatic cancer cells are dying 48 hours after infection with vesicular stomatitis virus. Green labels the cell membrane, blue is the cell DNA, and red is virus. Image taken using 40X objective with confocal Olympus FluoView 1000 microscope using filters for DAPI (blue), FITC (green), and Alexa Fluor 568 (red).



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Visualization of mitochondrial membrane potential and reactive oxygen production in live endothelial cells

Black background and divided into two halves. The left half shows multiple green teardrop-shaped cells with round dark centers, indicative of the nucleus of the cell. The right half shows the same cells but the green coloring now appears diffuse and is significantly brighter, indicative of mitochondrial membrane depolarization.

Samantha A. Pennington, Mark G. Clemens, Ph.D. Department of Biological Sciences
Recent studies have suggested that function of the endothelial cells lining blood vessels may be dependent upon changes in function of the mitochondria which produce energy for the cell. In order to be able to visualize mitochondrial function in living cells, human microvascular endothelial cells (HMECs) were visualized at a magnification of 40X using an Olympus FV500 confocal microscope following staining with multiple fluorescent dyes. Rhodamine 123 (Rh123, green) is a positively-charged stain that partitions into electronegative areas of the cell, such as polarized mitochondria so that the intensity of fluorescence is proportional to the mitochondrial membrane potential. Dihydroethidium (DHE, red) fluoresces only if it has been oxidized to ethidium, making it useful for detecting reactive oxygen species (ROS) production. Hoechst dye stained DNA to visualize nuclei. With these we studied the effects of mitochondrial uncoupling. Images were captured before and after treatment with FCCP, a mitochondrial uncoupler. FCCP caused rapid loss of Rh123 from the fibrillary mitochondria. DHE showed an uncoupler-induced mitochondrial ROS production following complete mitochondrial membrane depolarization. Results demonstrate that uncoupling and production of ROS in the mitochondria can be separated and mitochondrial depolarization occurs in an all-or-none fashion.



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In vivo high resolution imaging of liver mitochondrial membrane potential

Dark background and is composed of numerous cells of predominantly pentagonal and hexagonal shaped with an intricate branching pattern that is green in color. Large round blue centers are present in most cells, indicating the position of the nucleus of the cell.

Samantha A. Pennington, Mark G. Clemens, Ph.D. Department of Biological Sciences
Although mitochondria have been long recognized to be important for providing energy for the cells, more recently they have been shown to be involved in many other aspects of cell regulation, especially in cell injury. A major factor for mitochondrial regulation is maintenance of a highly electronegative membrane potential. While imaging approaches have been used for isolated cells, in vivo imaging of these structures which are as little as 0.2 µm in length, remains a challenge. This image demonstrates our development of a method for imaging individual mitochondria in the liver of live anesthetized mouse. Rhodamine 123 (Rh123, green) was infused intravenously and Hoechst stain (to stain cell nuclei) was loaded by surface exposure. Mice were then placed on an Olympus FV500 confocal microscope for imaging. The image was captured using a 100x oil immersion objective. Individual hepatocytes are identified by their blue nucleus and clear borders. In cells where no nucleus is observed, it is below the plane of focus. Importantly, individual mitochondria can be clearly observed. This indicates that it is feasible to visualize individual mitochondrial function in the livers of intact living animals allowing their study in complex disease states.



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Confocal in vivo imaging of hepatic blood flow and metabolic regulation

Dark background and shows numerous red oval to rectangular shaped cells connected to each to form the blood vessels of the liver. Vitamin A droplets appear blue-violet in color and are randomly dispersed throughout the image.

Samantha A. Pennington, Mark G. Clemens, Ph.D. Department of Biological Sciences
Altered blood flow regulation in the liver is an important contributor to liver injury during systemic infection (sepsis). Regulation of blood flow is complex, involving many cell types. Because of this complexity, in vivo studies are required. This is complicated by the fact that critical cellular responses are heterogeneous and occur in very small areas. In vivo imaging offers a potential means to study blood flow, metabolic responses and the contribution of multiple cell types simultaneous, but requires the development of high resolution, multimodal methods. This image demonstrates the results of our recent efforts to enhance in vivo imaging of both structures and metabolic status. A mouse was anesthetized with isoflurane and the liver exposed on an Olympus FV500 confocal microscope with 40x objective. Mitochondrial membrane potential as an indicator of metabolic status was imaged with tetramethylrhodamine methyl ester (TMRM, red). Tissue phagocytes were identified by uptake of green fluorescent beads and hepatic stellate cells which regulate the diameter of the sinusoids (hepatic capillaries, S) were identified by the blue fluorescence of vitamin A droplets in the cells. This demonstrates the feasibility of simultaneously assessing multiple blood flow and metabolic parameters in the liver using high resolution confocal imaging.



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Breast Cancer Cells

Black background and shows a blue-gray slightly irregular round field covered with very small breast cancer cells colored pink-red.

Kenya Joseph1, PI: Kirill Afonin, Ph.D. 2 1Department of Biological Sciences, 2 RNA Nanotechnology Lab
This image was taken from our Leica TCS SP2 Confocal Laser Scanning Microscope at 10X magnification by a Samsung Galaxy 5 cell phone. Our lab creates RNA nanoparticles for use in cancer research, personalized medicine and targeted drug delivery. This image is of breast cancer cells of cell line MDA-MB-231. We use them to study siRNA nanoparticle uptake and silencing of GFP fluorescence expression in these cells and other nanoparticle uptake. At the point of this image, the cells were being cultured into a new passage and we were observing confluence or cell growth and adherence prior to splitting them into a new culture. Cancer cell lines are immortal and are used extensively in many areas of biological research to observe cellular processes and conditions. They are cells descended from a single cell and contain the same genetic makeup as their original tissue source.



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Microparticles harboring sonic hedgehog modulates the endothelial cytoskeleton function in inflammation via rho kinase pathway

Four separate images, all of which are composed of numerous cells that line the inside of blood vessels. The cells are stained red with a blue circular nucleus in the center of most of the cells. Image A shows these cells with small fibers located mostly at the periphery of the cells. Images B and C show how these fibers are redistributed throughout the cells. Image D shows a significant decrease in the number of fibers and the cells have a more moth-eaten appearance

Neha Attal (PhD student) and Dr. Mark Clemens (Advisor) Department of Biological Sciences
All blood vessels are lined by endothelial cells. These regulate the diameter (blood flow) and the leakiness of the vessel. During inflammation, these cells are altered. One potential mediator is a protein called sonic hedgehog (shh) which is release in vesicles called microparticles (MP) from injured cells. Our goal is to determine the role of shh in altering endothelial cytoskeleton function during inflammation. This regulates the shape of the cell as well as its ability to regulate blood flow. To test this, we used human microvascular endothelial cells (HMECs) in culture. The cells were incubated with MP containing shh and then the cytoskeleton was stained with red fluorescent phalloidin, a toxin that binds to the actin in the cytoskeleton and the nucleus with DAPI which binds to DNA and then imaged on an Olympus FV500 confocal microscope. In normal cells (A), actin is mainly located at the edge of the cell. MP (B) or a shh agonist (C) treatment causes the actin to reorganize into "stress fibers". This is thought to be regulated by a cellular regulator rho kinase. When an inhibitor of rho kinase is used (D), stress fiber formation is largely prevented confirming the role of rho kinase.



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Isolated Lymphocytes

Black background and shows a perfectly round tri-color (red, yellow, blue) field covered with cells of the immune system colored blue.

Kenya Joseph (undergraduate) 1, PI: Kirill Afonin, Ph.D. 2 1Department of Biological Sciences, 2RNA Nanotechnology Lab
This image was taken from our Leica TCS SP2 Confocal Laser Scanning Microscope at 10X magnification by a Samsung Galaxy 5 cell phone. Our lab creates RNA nanoparticles for use in cancer research, personalized medicine and targeted drug delivery. Part of our research encompasses observing how these nanoparticles behave in human cells and my particular project is observing and elucidating the mechanism for cellular entry of the nanoparticles in human lymphocytes or white blood cells taken from whole blood. After I isolate the white blood cells from whole blood, I use the microscope to confirm the presence of live cells prior to introducing our fluorescently tagged nanoparticles. We then use flow cytometry to observe the entry of the particles in the lymphocytes. The image was edited artistically using filters only in Adobe Photoshop.



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Isolated Primary Colors

Four round fields horizontally connected to one another. The first is yellow, the second is red, the third is blue, and the fourth is green. All have very small black cells of the immune system on their surfaces.

Kenya Joseph1, PI: Kirill Afonin, Ph.D. 2 1Department of Biological Sciences, 2 RNA Nanotechnology Lab
These four images were taken from our Leica TCS SP2 Confocal Laser Scanning Microscope at 10X magnification by a Samsung Galaxy 5 cell phone. Our lab creates RNA nanoparticles for use in cancer research, personalized medicine and targeted drug delivery. Part of our research encompasses observing how these nanoparticles behave in human cells and my particular project is observing and elucidating the mechanism for cellular entry of the nanoparticles in human lymphocytes or white blood cells taken from whole blood. After I isolate the white blood cells from whole blood, I use the microscope to confirm the presence of live cells prior to introducing our fluorescently tagged nanoparticles. We then use flow cytometry to observe the entry of the particles in the lymphocytes. The image was edited artistically by combining four pictures taken successively using filters only in Adobe Photoshop.



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MB-MDA 231 nonGFP Cells

Multi-colored round field with numerous small dark green round and branched breast cancer cells across the surface. A curved red band lies in close proximity to the field.

Kenya Joseph1, PI: Kirill Afonin, Ph.D. 2 1Department of Biological Sciences, 2 RNA Nanotechnology Lab
This image was taken from our Leica TCS SP2 Confocal Laser Scanning Microscope at 10X magnification by a Samsung Galaxy 5 cell phone. Our lab creates RNA nanoparticles for use in cancer research, personalized medicine and targeted drug delivery. This image is of breast cancer cells of cell line MDA-MB-231. We use them to study siRNA nanoparticle uptake and silencing of GFP fluorescence expression in these cells and other nanoparticle uptake. At the point of this image, the cells were being cultured into a new passage and we were observing confluence or cell growth and adherence prior to splitting them into a new culture. In this image, many cells have died and only a few survive here. Cancer cell lines are immortal and are used extensively in many areas of biological research to observe cellular processes and conditions. They are cells descended from a single cell and contain the same genetic makeup as their original tissue source.



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Moon or Lymphocytes

Black background and shows a large round multi-colored field. Its surface has a rough texture and is covered with cells of the immune system colored blue.

Kenya Joseph1, PI: Kirill Afonin, Ph.D. 2 1Department of Biological Sciences, 2 RNA Nanotechnology Lab
This image was taken from our Leica TCS SP2 Confocal Laser Scanning Microscope at 10X magnification by a Samsung Galaxy 5 cell phone. Our lab creates RNA nanoparticles for use in cancer research, personalized medicine and targeted drug delivery. Part of our research encompasses observing how these nanoparticles behave in human cells and my particular project is observing and elucidating the mechanism for cellular entry of the nanoparticles in human lymphocytes or white blood cells taken from whole blood. After I isolate the white blood cells from whole blood, I use the microscope to confirm the presence of live cells prior to introducing our fluorescently tagged nanoparticles. We then use flow cytometry to observe the entry of the particles in the lymphocytes. The image was edited artistically using filters only in Adobe Photoshop.



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The Crack

Complete crack into carbon material, which has a course texture and a grid pattern. Numerous small latex spheres, measuring about 260 nanometers in diameter, are sitting on top of the material and inside the crack.

Shank Kulkarni Department of Mechanical Engineering and Engineering Science
Image taken from: Transmission electron microscopy (Grigg 147) Crack under microscope. Black circles are latex spheres.



Electrical and Computer Engineering

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Photoluminescence imaging of electrons in a semiconductor - similarity between a microscopic and macroscopic world

Two panels, each with a three-dimensional coordinate system. The left panel shows the surface of the XY axis colored a shade of purple with a cone shape generated in the center. The base of the cone is blue. As it rises, the colors change to green, yellow, orange, and red, illustrating an increase in height of the cone, which indicates more light intensity. In the right panel, the top surface of the XY axis is colored bright orange with an upside-down cone emerging from the center. As the cone stretches downwards, the colors change from yellow, green, blue, and then to purple, illustrating an increase in depth of the cone, which indicates less light intensity

Yong Zhang, Ph.D. Department of Electrical and Computer Engineering
Left panel: a tightly focused laser beam (with a diameter about 0.7 µm) is used to illuminate a GaAs semiconductor. At the illumination site, a large number of electrons are lifted to excited states by the photon energy. These electrons will diffuse to the surrounding area because of the concentration gradient. In the meantime, they will fall back to the ground state by emitting light. By imaging the spatial distribution of the emitted light intensity, one can obtain the spatial distribution of the electrons, and extract a material parameter called electron diffusion length that is a measure of the material quality. The large the value is the better the material quality. This phenomenon is very much similar to pooling down water continuously on a surface to watch how far the water can spread, depending on the characteristic of the surface. Right panel: an alternative way to measure the electron diffusion where the excited state electrons are generated by light in an equal number everywhere, and they are expected to emit light with a uniform intensity. However, near a defect, electrons within the electron diffusion length from the defect site will be pulled into the defect and depleted without giving out light, resulting in a dark region surrounding the defect. One can also derive electron diffusion length by analyzing the light intensity distribution in the vicinity of the defect. This phenomenon is similar to the water flow pattern near a sink, where the impact range of the sink depends on the characteristic of the surface. Both measurements were performed using an optical spectroscopy system equipped with a confocal optical microscope in Prof. Yong Zhang’s lab where his group are carrying out a wide variety of research projects related to optoelectronic materials and devices (such as solar cells, LEDs, photo-detectors) as well as fundamental sciences.



Kinesiology

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Aged Mouse Skeletal Muscle Fiber Types

Black background and shows an irregularly round piece of hindlimb muscle from a mouse. This muscle is composed of many cells of a variety of shapes. Most of the cells are colored green, while a few are blue, black, and red.

Marcus M. Lawrence1, Joshua R. Huot1, Kevin A. Zwetsloot2, R. Andrew Shanely2, Scott. E. Gordon1, Susan T. Arthur1. 1)Department of Kinesiology, 2)Department of Health and Exercise Science, Appalachian State University
The image was captured at 10x magnification on an Olympus IX71 Inverted Fluorescent Microscope equipped with a DP71 Digital Camera. This image displays skeletal muscle fiber (i.e., cell) types (SkMFT) of an aged 21 month old (equivalent of ~65 human years) male mouse Plantaris (a muscle from the hindlimb) cross-section cut at a thickness of 10µm. SkMFT are found in a continuum as follows: type I → IIA → IIX → IIB, with type I being the smallest and lowest force producing fiber and type IIB being the largest and highest force producing fiber. The SkMFT in the image were fluoresced using antibodies directed to the following: type I(red), IIA(blue), IIX(black), IIB(green), and laminin(red; connective tissue around each fiber). The age-related loss of skeletal muscle mass and function (i.e., sarcopenia) displays a preferential loss of type II (including IIA, IIX, and IIB) fibers leading to decrements in whole muscle size and strength which can eventually lead to institutionalizations and mortality. The goal of examining SkMFT in the current research project was to test whether 28-day supplementation with a compound produced from the plant Ajuga turkestanica can enhance type II fiber numbers and/or size beyond a control treatment in 21 month old male mice.



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Effects of Notch Inhibition on Aged Skeletal Muscle Repair Following Downhill Running - Control mouse tissue; no DHR (1)

Cross-section of skeletal muscle fibers, that are dark in color and mostly pentagonal and hexagonal shapes. Located around the periphery of the fibers are small nuclei of varying shapes, represented in blue. Also around the periphery, is another protein important for muscle tissue development, which is red in color.

Joshua Huot, Cheaslei Weathers, Susan Tsivitse Arthur, Ph.D. Department of Kinesiology
This image represents the control condition of the investigation and shares the same description as the inhibition mouse tissue in the image below. Please refer to the next image for the description.



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Effects of Notch Inhibition on Aged Skeletal Muscle Repair Following Downhill Running : Inhibition mouse tissue - 6 days post-DHR (2)

Cross-section of mouse gastrocnemius, or calf muscle, showing many skeletal muscle fibers. The fibers appear on a spectrum from dark green to nearly black and are mostly rounded pentagonal and hexagonal shapes. Located around the periphery of the fibers are small nuclei of varying shapes, represented in blue. There is additional protein around the periphery that is important in muscle tissue development, which is mostly green, with some dark red

Joshua Huot, Cheaslei Weathers, Susan Tsivitse Arthur, Ph.D. Department of Kinesiology
Left image: Control mouse tissue; no DHR Right image: Inhibition mouse tissue, 6 days post DHR One of the key contributors to sarcopenia (the age-related loss of muscle mass) is a dysfunctional repair process of aged muscle following injury. The aim of this study was to observe the role of Notch signaling on Wnt signaling and overall repair capacity in aged skeletal muscle following an injurious bout of downhill running. Immunofluorescence staining was performed on sections of mouse skeletal muscle to observe differences in proteins that contribute to the skeletal muscle repair process. The mouse tissue was tagged with primary antibodies overnight, followed by secondary fluorescent antibodies yielding the above imaging of nuclei (blue), myoD (red: a key skeletal muscle repair factor), and Lef1 (green: a protein of Wnt signaling that contributes to the later stages of the repair process).The image above was captured on an Olympus IX71 inverted fluorescent microscope (40x). This demonstrates the importance of Notch signaling for proper skeletal muscle repair following injury. Inhibiting Notch signaling (picture on the right) reduced the overall repair capacity of aged skeletal muscle (myoD: red) compared to a control condition (picture on the left), while at the same time increasing Wnt signaling (Lef1: green).



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Protein Kinase C-Θ Regulates Insulin Sensitivity in Skeletal Muscle Cells

Cross-section of many skeletal muscle fibers of varying shades of red and are irregular in shape, but mostly rounded pentagonal and hexagonal shapes. Located around the periphery of the fibers are small nuclei of varying shapes, which fluoresce blue.

Bailey Peck and Joseph Marino Laboratory of Systems Physiology, Department of Kinesiology
Understanding the mechanisms in which high fat diets induce insulin resistance, has led to the investigation of cytoplasmic proteins activated by lipid metabolites. Protein Kinase C Theta has been implicated in the inhibition of insulin signaling in the presence of diacylglycerol. Using immunofluorescence to identify activated Protein Kinase C theta using mouse PKC theta primary antibody, followed by a goat anti-mouse Texas red fluorescent secondary antibody demonstrates the protein content of PKC theta in the Gastrocnemius of a 15 week old high fat fed mouse (C57BL6/J). DAPI was used to identify the nuclei in the periphery of the muscle fibers which fluoresces blue. This image was shot at 100x using an Olympus XI71 microscope with a fluorescent bulb.



Mechanical Engineering and Engineering Science

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Chatter on Ultraprecision Diamond Turned Surface

Spiral of gradations of yellow to orange layers with a sunburst in the center. The image scale represents 160 micrometers long on the X axis and 160 micrometers long on the Y axis. The depths of the spiral layers vary from approximately 0.005 to 0.015 micrometers in height.

John Troutman Department of Mechanical Engineering and Engineering Science
Optical components for infrared applications are often produced by ultraprecision diamond turning, where a single-point diamond tool is used to remove material from a part until the desired surface is generated. During such a cutting process, “chatter”, or undesired vibration between the cutting tool and part, can produce irregular patterns on the part surface and poor finish. This image is a height map of the face of a piece of glass, diamond-turned under chatter conditions. The data was acquired with a Zygo ZeGage Scanning White Light Interferometer (SWLI) using a 50x Mirau-type interferometric objective and 1x field zoom. The 1024 x 1024 pixel height map was plotted with MATLAB.



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Electron Paths

Series of three pictures from left to right: (a) horizontal light and dark blue small triangles connected in a fairly consistent pattern forming several rows (b) seven rows of transverse linear dark blue waves following a downhill direction (c) transverse light and dark blue small triangles connected in a fairly consistent pattern forming several rows following an uphill direction.

Jimmie Miller Ph.D. William States Lee College of Engineering, Mechanical Engineering and Engineering Science Department
The image series visualizes 1 nm X 1 nm raster scans from a tunneling microscope of the surface electron states of graphite with and without a transverse surface current flowing. (a) reveals a normal state with the bright regions representing (some) atomic position sites. (b) when a transverse surface current is induced the electron paths are shown to be along preferred directions indicating the direction of current flow. (c) reveals the surface electronic state after the transverse surface current have ceased with a return to a normal state. The investigation was part of research to determine the feasibility of creating a digital memory with information bits the size of nanometers. Digital Instruments Nanoscope II scanning tunneling microscope (STM) in constant height mode. An atomically sharp probe is raster scanned across the surface of a material with a small applied voltage between the material and the probe inducing electron flow between the material and probe. The current flow magnitude depends on the electron probability density and potential barrier at each position. Higher current flows are visualized with light coloring while the darker regions have a lesser flow.



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A little forest?

Gray scale image shows many cut ribbons of thin metal of varying shades. The long ribbons lay vertically and most of the shorter pieces have fallen horizontally.

Eric Browy (MS 2014), Supervisor: Chris Evans Department of Mechanical Engineering and Engineering Science
This scanning electron micrograph (2300X) shows material removed in ultra-precision machining where a single crystal tool diamond removes material layers less than 5 micrometers thick. Diamond is the hardest material and should cut anything – except that chemistry gets in the way. Some very hard metal cause no tool wear and some soft ones very rapid wear. Our overall research goal is to improve economic utilization of expensive machines and tools; understanding and predicting the chemical wear of diamond tools as a function of chemical composition of the workpiece will eliminate trial and error methods in developing new processes. The shapes of the material removed provides information on the process.



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Morning dew on nanowires

Dark blue background with bright and light blue linear projections and knob-like structures across the top half of the image. It resembles dew drops on thin blades of grass.

Siang Yee Chang, Terry T. Xu, Ph.D. Department of Mechanical Engineering and Engineering Science
Image credit: Dr. Zhe Guan, alum of Xu group, now at Intel Corporation, AZ. Formation of silicon oxide (SiOx) nanoballs along the boron carbide (B4C-type) nanowires in a horizontal tube furnace. These hybrid nanostructures were grown on a SiO2/Si substrate. They were formed when boron, boron oxide and carbon powders were used as the reaction precursors. At elevated temperatures, the SiOx molecules presented in the vapor state. As the furnace temperature decreased, the saturated vapor pressure of SiOx molecules reduced, leading to the nucleation of SiOx molecules along the B4C nanowires. Microscope used: JEOL JSM 6480 SEM (Scanning Electron Microscope). Magnification: 9,000X Methods used: Colored with Adobe Photoshop CS6



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Nanocoral

Purple image shows a variety of tentacle-like structures extending from the background, resembling coral in the depth of the oceans.

Siang Yee Chang, Terry T. Xu, Ph.D. Department of Mechanical Engineering and Engineering Science
Image credit: Dr. Zhe Guan, alum of Xu group, now at Intel Corporation, AZ. The image shows the growth of flower-like boron-based nanostructures on the sapphire substrate. These nanostructures were synthesized in the low-temperature regions of a low pressure chemical vapor deposition (LPCVD) system during co-pyrolysis of diborane (B2H6) and methane (CH4) at elevated temperatures. Microscope used: JEOL JSM 6480 SEM (Scanning Electron Microscope). Magnification: 10,000X Methods used: Colored with Adobe Photoshop CS6



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The best of nature - green grass & blue sky

Solid blue background with dark and light green linear projections stretching up from the base. The image appears to look like blades of grass against a blue sky.

Siang Yee Chang, Terry T. Xu, Ph.D. Department of Mechanical Engineering and Engineering Science
Image credit: Dr. Zhe Guan, alum of Xu group, now at Intel Corporation, AZ. The image shows the growth of boron carbide (B4C-type) nanowires at the edge of the sapphire substrate. The nanowires were synthesized by co-pyrolysis of diborane (B2H6) and methane (CH4) gases at 1050°C in a low pressure chemical vapor deposition (LPCVD) system. Microscope used: JEOL JSM 6480 SEM (Scanning Electron Microscope). Magnification: 10,000X Methods used: Colored with Adobe Photoshop CS6



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When autumn comes...

Green background with a roughened texture. Two rose-colored puffy ball-like structures appear to extend forward to sit on top of the background, as if to have a bird's eye view of colorful autumn leaves on the grass.

Siang Yee Chang, Terry T. Xu, Ph.D. Department of Mechanical Engineering and Engineering Science
Image credit: Dr. Zhe Guan, alum of Xu group, now at Intel Corporation, AZ. The α-tetragonal boron nanoplatelets grow into a puffy-ball-like structure on the SiO2/Si substrate surface during the synthesis of boron-based one-dimensional (1D) nanostructures. The synthesis involves the co-pyrolysis of diborane (B2H6) and methane (CH4) gases at elevated temperatures in a low pressure chemical vapor deposition (LPCVD) system. These puffy-ball-like structures are usually found in the low temperature regions (630 - 750 ℃) within the LPCVD system. Microscope used: JEOL JSM 6480 SEM (Scanning Electron Microscope). Magnification: 100X Methods used: Colored with Adobe Photoshop CS6



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Colorful nanoribbons

Blue background with bright rainbow-colored (multiple colors) twisted ribbons prominently displayed in the foreground.

Siang Yee Chang, Terry T. Xu, Ph.D. Department of Mechanical Engineering and Engineering Science
Image credit: Dr. Zhe Guan, alum of Xu group, now at Intel Corporation, AZ. Formation of twisted boron nanoribbons on the Si/SiO2 substrate during the synthesis of boron carbide one-dimensional (1D) nanostructures by co-pyrolysis of diborane (B2H6) and methane (CH4) gases at elevated temperatures in a low pressure chemical vapor deposition (LPCVD) system. Microscope used: JEOL JSM 6480 SEM (Scanning Electron Microscope). Magnification: 3,000X Methods used: Colored with Adobe Photoshop CS6



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Circles

Dark image with four circles of increasing diameter, each nested within the other, and the largest measuring about 150 nanometers in diameter. The smallest has a bright white perimeter. A thin rectangular beam lies partially across the circles.

Shank Kulkarni Department of Mechanical Engineering and Engineering Science
Image taken from: Transmission electron microscopy (Grigg 147). Image shows diffraction patterns for Au (gold) on holey carbon film. Beam blocker is used to avoid direct beam.



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Latex Globules

Gray background with irregular latex spheres connected to each other. The main gray sphere measures approximately 260 nanometers in diameter.

Shank Kulkarni Department of Mechanical Engineering and Engineering Science
Image taken from: Transmission electron microscopy (Grigg 147). Image shows very tiny latex globule which is simple attached to one other latex globule. Deformations on such a small scale are really beautiful.



Nanoscale Science

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Absorb the Spectrum

Blue background with numerous mostly circular particles with a green hue and pink backdrop. Some particles and clusters of particles are colored bright red and yellow. Particles range from 25 to 50 nanometers in diameter.

Kathleen Dipple, Assisted by: Andrew Tobias Nanoscale Science Ph.D. Program
A transmission electron microscope (TEM) at a 50 K magnification was used to generate the image. The program, Image J was used to alter the colors of the image. The image is of gold/silver sulfide nanoparticles, which can be used for improved photovoltaic devices. The scale bar is 100 nm.



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Nano Cyan

Mottled green-blue background with a large cluster of mostly circular particles, which are green-blue to dark blue in color. Most particles measure about 30 nanometers in diameter.

Kathleen Dipple, Assisted by: Andrew Tobias Nanoscale Science Ph.D. Program
A transmission electron microscope (TEM) at a 120 K magnification was used to generate the image. The program, Image J was used to alter the colors of the image. The image is of gold/silver sulfide nanoparticles, which can be used for improved photovoltaic devices. The scale bar is 50 nm.



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Green Glow at 100k

Faint green background with a large cluster of mostly circular particles, which are green to dark green. Most particles measure about 30 nanometers in diameter.

Kathleen Dipple, Assisted by: Andrew Tobias Nanoscale Science Ph.D. Program
A transmission electron microscope (TEM) at a 100 K magnification was used to generate the image. The program, Image J was used to alter the colors of the image. The image is of gold/silver sulfide nanoparticles, which can be used for improved photovoltaic devices. The scale bar is 50 nm.



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15 Nanometer Shell

Burnt orange background with approximately 23 particles, mostly circular in shape with black interiors and outlined in purple, which is feathered around the circumference of each particle. The larger particles measure about 15 nanometers in diameter.

Kathleen Dipple, Assisted by: Andrew Tobias Nanoscale Science Ph.D. Program
A transmission electron microscope (TEM) at a 120 K magnification was used to generate the image. The program, Image J was used to alter the colors of the image. The image is of gold/silver sulfide nanoparticles, which can be used for improved photovoltaic devices. The scale bar is 50 nm.



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Awareness

Mottled pink background with a large cluster of mostly circular particles, which are dark pink to purple. Most particles measure about 30 nanometers in diameter.

Kathleen Dipple, assisted by: Andrew Tobias Nanoscale Science Ph.D. Program
A transmission electron microscope (TEM) at a 100 K magnification was used to generate the image. The program, Image J was used to alter the colors of the image. The image is of gold/silver sulfide nanoparticles, which can be used for improved photovoltaic devices. The scale bar is 50 nm.



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Cyanzoomed

Mottled green-blue background with a large cluster of mostly circular particles, which are green-blue to dark blue in color. Most particles measure about 30 nanometers in diameter.

Kathleen Dipple, Assisted by: Andrew Tobias Nanoscale Science Ph.D. Program
A transmission electron microscope (TEM) at a 100 K magnification was used to generate the image. The program, Image J was used to alter the colors of the image. The image is of gold/silver sulfide nanoparticles, which can be used for improved photovoltaic devices. The scale bar is 50 nm.



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Infrared

Yellow background with a large cluster of mostly circular particles, which are reddish orange to dark brown in color. Most particles measure about 30 nanometers in diameter.

Kathleen Dipple, Assisted by: Andrew Tobias Nanoscale Science Ph.D. Program
A transmission electron microscope (TEM) at a 100 K magnification was used to generate the image. The program, Image J was used to alter the colors of the image. The image is of gold/silver sulfide nanoparticles, which can be used for improved photovoltaic devices. The scale bar is 50 nm.



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Nanoparticles

Faint blue background with numerous mostly circular particles with a blue-green hue. Some particles and clusters of particles are colored blue-green to very dark blue. Particles range from 25 to 50 nanometers in diameter.

Kathleen Dipple, Assisted by: Andrew Tobias Nanoscale Science Ph.D. Program
A transmission electron microscope (TEM) at a 50 K magnification was used to generate the image. The program, Image J was used to alter the colors of the image. The image is of gold/silver sulfide nanoparticles, which can be used for improved photovoltaic devices. The scale bar is 100 nm.



Physics and Optical Science

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Motley Silicon Wafer

Black background showing a vertical rectangular structure with few vertical ridges on the right side and deep horizontal ridges on the left. The structure's surface has a radiance of green, purple, pink, and yellow colors.

Zeba Naqvi, Tsing-Hua Her, Ph.D. Department of Physics and Optical Science
Plasma Enhanced Chemical Vapor Deposition (PECVD) is designed to coat flat wafers. However we used PECVD to coat optical fibers after modifying the tool. This modification changed the optical properties of the deposited materials. Now we had to characterize the new properties. A conventional way to do the characterization is ellipsometry, but it needs a flat sample unlike an optical fiber. Therefore we tried to coat small pieces cut out of Si wafer in the tool modified for optical fibers. This image is of one such piece. Unfortunately it was found that the properties on the wafer were different from that on fiber and this exercise of using a wafer did not help. Nevertheless it gave a beautiful image that is my desktop background since then. This is a dark field image from Olympus BX51 of the edge of the wafer piece coated with Silicon Nitride. Magnification is 10x. These are the natural colors due to thin film interference. No additional effects have been added.



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Beauty in all things, including failure

Black background with numerous irregular circles, globs, and distorted shapes with both straight and rounded edges. The interior of most of the shapes is gray with many having layers of various shades of blue, green, yellow, purple, and orange colors diffusing throughout the shape, some in concentric circles

Meng Liu, Thomas Suleski, Ph.D. Department of Physics and Optical Science
This is a real-color image of the surface of a specialized (LaSFn9) glass blank after an (unsuccessful) attempt to chemically clean the glass blank in preparation for fabrication of a very precise optical component. The component was to be used as a calibration artifact for a new, non-invasive optical process for measurement of the tear-film thickness of the human eye. The cleaning procedure used resulted in small-scale fracturing of the top layer of the glass. All colors in the microscope image arise from optical interference effects within very thin films of air underneath fragmented sections of the glass surface. The thin-film interference effects are conceptually very similar to those seen in oil films on a puddle on a rainy day, or in a soap bubble. The image was taken on an Olympus LEXT OLS4000 3D Laser Confocal Microscope in Grigg Hall at 20X nominal magnification.



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Cross or Sword

Gray background with intermittent stippling. In the center is a narrow shape in the outline of an inverted cross or upright sword.

Aaron Brettin1, Dr. Vasily Astratov1, Dr. Yuri Nesmelov1, and Dr. Kenneth Allen2
1) Department of Physics and Optical Science, 2)Georgia Tech Research Institute - Advanced Concepts Laboratory
This image was captured using a Mitutoyo upright microscope using an M plan Apo NIR objective of 10x with a numerical aperture of 0.26. These were uniquely formed when a protein sample was prepped using a salt solution. This knowledge is being used to alter the preparation methods of the protein solution.