Flim wallpapers

To deduce the FRET efficiency, without the need to measure the emission lifetime of the acceptor mCherry. The main objective of the current proposal is hence for the PI to obtain in-depth training in optics and fluorescence, particularly FLIM and its associated instruments and imaging analysis. Part of the training requires the PI to attend formal courses and training programs whereas the other aspects of training involve more self-studying and interactions with collaborators. Accordingly, three specific aims are proposed for the research plan: (1) develop a mOrange2/mCherry-based FAK biosensor and characterize it by FLIM;(2) visualize the spatiotemporal map of FAK activity under different flows with FLIM;(3) simultaneously monitor FAK and Src activities under different flows with FLIM. The information obtained will significantly advance our systematic understanding of the molecular mechanism by which different flows affect cardiovascular diseases, such as atherosclerosis and restenosis. The newly developed biosensors will also provide powerful tools for detecting cardiovascular diseases and assessing the efficacy of therapeutic inhibitors.Public Health RelevanceEndothelial cells (ECs) are continuously exposed to flow and its resultant shear stress, which plays crucial roles in regulating EC function and the ensuing patho-physiological processes, such as atherosclerosis and restenosis. This proposal will allow the PI to obtain in-depth training in optics and fluorescence, particularly FLIM and its associated instruments and imaging analysis, such that the PI can integrate FRET and FLIM to visualize FAK and Src activities simultaneously in live ECs to advance our systematic understanding of the molecular mechanism of mechanotransduction. Therefore, the success of the proposed project will have significant impact on improving public health.

The Integration of FLIM and FRET for the Visualization of Flow-induced Molecular Hierarchies The research activity of the PI's laboratory is focused on the integration of cutting edge technologies for the elucidation of molecular mechanism in vascular biology, with the ultimate goal of enhancing our knowledge on cardiovascular functions and contributing to the health and well-being of humankind. In particular, novel imaging biosensors and technologies will be developed and employed to visualize the signaling transduction with high spatiotemporal resolutions in live cells under different mechanical forces, e.g. shear stress. In the long run, we aim to apply the knowledge and insights gained via these novel technologies for the development of new strategies and reagents to treat cardiovascular diseases, including thrombosis and atherosclerosis. Along this line, we proposed to apply biosensors based on fluorescence resonance energy transfer (FRET) to visualize the Src activity and its related molecular hierarchy at sub-cellular levels during mechanotransduction, which is funded by NHLBI (R01HL098472). Since FAK activity and its autophosphorylation at tyrosine 397 are critical for the Src activation, we plan to investigate the spatiotemporal coupling between FAK and Src activations in endothelial cells under mechanical stimulation. Recently, we have developed a new FRET pair, mOrange2 and mCherry, which contains colors distinct from the popular FRET pair, CFP and YFP. As such, two different molecular events, such as the activities of FAK and Src, can be simultaneously monitored in the same live cell with biosensors based on mOrange2/mCherry and CFP/YFP. However, there is a certain overlap between the excitation spectra of mOrange2 and mCherry to cause a non-specific cross- excitation of mCherry, which can cause artifacts and lower the sensitivity of FRET biosensors. Fluorescence lifetime imaging microscopy (FLIM) could help to solve this problem because the emission lifetime measurement of the donor mOrange2 alone is sufficient

Listening to Granny Smith; 17:22, 19:04 - in the stands; 19:21 - cheering for Berryshine and "Piña Colada"Duplicates: 19:21 - two in the crowd6None7None83:13, 3:27, 3:33, 4:31, 4:52 - in the crowd praising Rainbow Dash; 17:56 - in the crowd at Mare Do Well's thank-you parade; 18:05 - gasps when Rainbow Dash confronts Mare Do Well910:18 - watching an airship's maiden voyage during Becoming Popular (The Pony Everypony Should Know)10None110:44 - flying through Canterlot; 1:51, 3:32, 3:46, 14:33, 20:05, 20:39 - in the pageant audience; 1:55 - handling a cloud prop backstage; 5:07, 5:36 - acting in the pageant; 20:24 - singing The Heart CarolDuplicates: 3:32, 14:33, 20:05, 20:39 - multiple; 3:46 - one behind Orange Swirl12None13None142:43 - at Applejack's sendoff party152:57 - drinking apple cider; 4:09 - in the crowd complaing; 5:24 ~ 8:40 - in the crowd during The Flim Flam Brothers; 8:50 ~ 12:53 - in the crowd; 10:36 - in line for apple cider; 13:10 - in the crowd at the cider-making competition; 19:49 - watching the Apples leave; 20:44 - watching Flim and Flam leave16None173:30 - in Ponyville Town Square185:35 ~ 6:14 - appears during Smile Song197:25 ~ 9:20 - at Iron Will's seminar (with multiple duplicates)204:55 ~ 5:04 - listening to Twilight and looking at Pinkie Pie21None.220:23 - next to White Lightning; 0:28 - outside Golden Oak Library; 2:22 ~ 3:34 - inside the library; 3:39 ~ 4:05, 7:10 ~ 10:02, 13:27 - on the training field; 14:39 ~ 20:18 - at the reservoir (with several duplicates)23None24None25None266:58, 8:17, 8:58 - looking at Queen Chrysalis; 20:28 - at the wedding receptionSeason three120:51 - among the ponies at the Crystal Faire211:43 - watching the jousting match (no wings or cutie mark)313:23 ~ 14:42 - in the crowd complaining about Pinkie PieDuplicates: 13:27, 14:17, 14:42 - one

Trixie to the remains of Ponyville to see if they can find remnants of the spell and undo it.They agree to go down there in the early morning, after Rainbow turns down idea to leave tonight. After everyone leaves Celestia then asks Raibow why she didn't want to do it a night. Rainbow then tells Celestia about the nightmares she's been having, to which Celestia then explains that she's been getting the same nightmares as well, she then explains that she thinks the nightmares are being done by Luna as way to try and contact them and that she believes she, as well as the other missing ponies, might still be alive.During the night, in a field near the Everfree forest, the Flim Flam brothers were sat around a fireplace, when both were attacked by what appeared to be a pegasus. The Pegasus is suddenly attacked and held down by Big Macintosh, who is then approached by Zecora who feeds the creature a potion. This cause the creature to writhe and spit out bloody chunks, as well as transform her back into a regular pegasus, revealing her to be Fluttershy.The Flim Flam brothers then introduces who they are and say they're looking for Ponyville. Zecora explains that Ponyville is gone and that they should leave as well. she then proceeds to lift the bloody chunks revealing them to be pieces of candy and throw them into the fire, where they explode into blue flames. Big Mac and Zecora then takes Fluttershy, who begins to wake up, before telling the brothers again they should leave, stating there are monsters lurking around out here.After the three leave, the brothers stay behind and start discussing what they should do, as a silver mist starts emitting from the forest. They both then hear laughter emitting from the mist, as something appears rustling in the grass. The brothers then decide to leave only to run into several candy ghouls, that chase them both into an abandon barn.Powers & AbilitiesEnhanced Immunity - As to be expected from a possibly immortal being, Candy Mare seems to be immune to most common diseases ordinary ponies could contract/spread. This would have most likely been caused by her transformation into becoming a candy-fused monstrosity as her body inside and outside mutated to become more resistant to Lemon Drop's experiments or potential risks.Enhanced Strength/Speed - Along with Candy Mare's increased immunity

• Photo Finish • Sapphire Shores • Fancy Pants • Jade Singer • Daring Do • Prim Hemline • Trenderhoof • Coloratura • Zesty Gourmand • Songbird SerenadeAntagonistsShadowbolts • Boy Bullies • Diamond Dogs • Flim and Flam • Queen Chrysalis • King Sombra • Lightning Dust • Big Boy and Runt • Sunflower • Mane-iac (henchponies) • Suri Polomare • Lord Tirek • The Dazzlings (Adagio Dazzle • Sonata Dusk • Aria Blaze) • North Star • King Longhorn • Principal Cinch • Wind Rider • Svengallop • Gladmane • The Storm King • Cozy Glow • Sludge • Princess Eris • Grogar • OpalineReformed: Gilda • Trixie • Discord • Ahuizotl • Garble • Larry • Sunset Shimmer • Snips and Snails • Buck Withers • Dr. Caballeron (henchponies) • Starlight Glimmer • Shadow Lock • Juniper Montage • Stygian • Tempest Shadow • Grubber • Wallflower Blush • Chancellor Neighsay • Vignette Valencia • Kiwi Lollipop and Supernova Zap • Sprout • Posey • MistyWonderboltsSpitfire • Soarin • Blaze • Surprise • Fire Streak • Misty Fly • Silver Zoom • Fleetfoot • High Winds • ThunderlanePhysiciansNurse Redheart • Dr. Horse • Dr. Fauna • Doctor Horse • Nurse poniesPillars of Old EquestriaStar Swirl the Bearded • Rockhoof • Mistmane • Flash Magnus • Somnambula • Mage MeadowbrookYoung SixSandbar • Gallus • Smolder • Yona • Ocellus • SilverstreamOther pony characters8-bit • All Aboard • Alphabittle • Angel Wings • Autumn Blaze • Barley Barrel • Blossomforth

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