Shown here is an image of an adult mouse colon. In green is keratin 18 and red is beta-catenin. Keratin 18 is part of the intermediate filament network that imparts mechanical strength to the epithelium of the colon. Beta-catenin is part of the adherens junction, which helps maintain cell-cell adhesion. Both cell adhesion and mechanical strength are necessary for proper maintenance of the colon tissue. The colon is made up of many crypts, which can be seen as long invaginations in longitudinal section, or as circles in cross-section. The crypts are the intestinal stem cell niche, which constantly replenish the epithelial cell population. The lifetime of an intestinal epithelial cell is about 3-7 days, so the crypt population of stem cells is necessary to maintain tissue homeostasis.
Third Place
Chaperone's Embrace
Graham Johnson & Sander Tans
The Scripps Research Institute & fivth.com
Optical tweezers reveal for the first time how the folding pathway of a protein (MBP, transparent) is altered by a chaperone (SecB, yellow). Here MBP is shown suspended in its molten globule state and destined for transport across the cellular membrane.
Second Place
Sea Creature Radiance
Michael Shribak
Marine Biological Laboratory
The diatom Arachnoidiscus have a silicified cell wall forms a pillbox-like shell (frustule) composed of overlapping halves that contain intricate and delicate markings. We used Olympus BX-61, 40x/0.75NA objective lens, consumer Nikon D40x SLR digital camera, and video-enchanced polarized light.
First Place
Save the Last Dance for me
Aswati Subramanian
Miami University
This is an image of the single cell organism Tetrahymena thermophila. This scanning electron image is taken with an accelerating voltage of 12 KeV and a working distance of 10mm.
CELLDANCE VIDEOS
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Honorable Mention
Zipping...
U. Serdar Tulu
Duke University
Focal adhesions are dynamic structures. They assemble and disassemble continuously. This movie shows this process happening in cells collectively during zipping process of dorsal closure in Drosophila embryo.
Drosophila embryo was imaged using spinning disc confocal microscopy and 100x lens. Focal adhesions are made visible with a GFP fusion protein in leading edge cells using UAS-Gal4 technology. Each frame is maximum intensity projection of a z-stack. The projected images were unsharp masked. Entire movie takes about 2 hours and 20 minutes. Marker bar shows 10 microns.
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Honorable Mention
Microdomaindance 2009
Emmanuel Derivery
Centre National de la Reherche Scientifique (CNRS)
WASH is a newly discovered Arp2/3 activator that triggers actin polymerization on a microdomain of early endosomes. This movie describes that these microdomains are highly dynamic at the surface of enlarged endosomes. Actin pymerization by Latruculin A treatment induces fusion of these microdomains, suggeting that Actin polymerization provides the force that allows segregation of distinct WASH microdomains. Mouse fibroblasts transiently expressing mCherry-Rab5aQ79L and stably expressing GFP-WASH were plated on glass-bottom culture dishes coated with fibronectin. Latrunculin was used at a final concentration of 1 µM. In this experiment, a pretreatment with Nocodazole (10 µM) was also performed to scatter endosomes and facilitate imaging. Cells were then imaged live using a custom spinning disc confocal microscope based on a Nikon TE2000-U inverted microscope, a 100X NA 1.45 oil immersion objective, a Yokogawa CSU22 spinning disk head and a CoolSnap HQ2 camera Photometrics). Several confocal planes were acquired (0.2 µm increment, 2 steps for first movie, 12 steps for second movie) then a maximum intensity projection was performed. Images were then processed with ImageJ. Bleaching was corrected and images from the GFP-WASH channel in the first movie were processed using a "à trous" wavelet filter created by Fabrice Cordelière. 3D reconstruction was performed using Amira. Animation was created using Adobe Flash and final movie was edited using Adobe Premiere.
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Third Place
To Paint a Van Gogh in 25 minutes
Dennis Breitsprecher
Hannover Medical School
The image series shows the spontaneous assembly of fluorescently-labeled actin monomers into filaments and bundles in presence of 1 µM of the actin severing protein cofilin. The loose, regular actin bundles display remarkable similarities to the world-famous paintings of Vincent van Gogh (although only 25 minutes were needed to produce them...).
Methods:
3 µM actin monomers (30% Oregon Green labeld on Cys 374) were polymerized in TIRF-buffer (50 mM KCl, 0,2 mM ATP, 1 mM MgCl2, 10 mM DTT, 50% methylcellulose, 20 mM Imidazole pH 7.3)in presence of 1 µM cofilin, and the time course of polymerization was recorded with an inverted IX81 Olympus TIRF-microscope equipped with an Orca E2 camera and an Olympus APON XO60XTIRF objective. The field corresponds to 100 x 140 µm, one frame corresponds to 20 sec. The image was processed using the ´rainbow RGB´ LUT from Image.
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Second Place
ATP Synthase
Graham Johnson
The Scripps Research Institute & fivth.com
Following a script written by Peter Walter (UCSF), this teaching animation created in 2001 by Graham Johnson describes the transfer of chemiosmotic energy into rotational energy, and ultimately into the chemical bond energy of ATP. The medical animator morphed between conformational states captured in the 3 fold crystal structure to illustrate the dynamics of this fascinating motor protein as accurately as possible but simplified the data considerably to make the visual information approachable to college-level audiences.
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First Place
2PEF Imaging Captures Gastrulation in Fly Embryos
Angela Stathopoulos
California Institute of Technology
Posterior and dorsal views of mesoderm cells as they spread to form a monolayer during gastrulation. This movement is essential for positioning the mesoderm to form heart and gut muscles. Cells are imaged live with a nuclear marker (H2A-gfp) using two-photon excited fluorescence (2PEF). The imaging data is then imported into data-processing software to segment and follow each nuclei over time (grey spheres=mesoderm nuclei). The ectoderm, which is the substratum for mesoderm cell migration, is shown for reference (green). Tracks are represented by lines with a temporal color code, where purple represents early time points and yellow represents late time points. The first view is dorsal, and the second view is posterior. Large grid mark = 20 µm.
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