Images from a scratch assay experiment at different time points. Human umbilical veinendothelial cells (HUVEC) were plated on gelatin-coated plastic dishes, wounded with ap20 pipette tip, and then imaged overnight using a microscope equipped with pointvisiting and live-cell apparatus. Scale bar = 120 μm.
The dish or chamber used to culture the cells should be carefully chosen to match theexperimental approach. Multi-well cell culture dishes (6, 12, or 24 wells) are a popularchoice because they are inexpensive and fabricated from plastic treated to encourage celladhesion. The wells in these dishes are also large enough to provide clearance formanipulating the monolayer as when creating a scratch wound or working with an insert. Forhigh-resolution imaging, as discussed below, it is necessary to use dishes with thin plasticor glass bottoms on the order of 160-190 μm (Number 1.5 coverslips). Although almostall cells will grow well on plastic, some cell types will not grow on glass unless it isspecially treated and coated with serum, gelatin or other extracellular components.
Scratch Live 1. 8 2
To simulate wounding, the most common approach is to create a gap by scratching a confluentmonolayer with a pipette tip, needle, or other sharp tool. This is known as the scratchassay and is a good choice because it is inexpensive and easy to implement. Usually, apipette tip is used to create a scratch one well at a time. As the scratch is createdmanually, it can be difficult to generate reproducible wounds. It is important to angle thepipette correctly as well as to apply consistent pressure to create a consistent gapwidth.37 In addition, applying toomuch pressure may damage the extracellular matrix, which can affect migration rates.28 While the scratch method is usuallyperformed one well at a time, some groups have scaled up the technique to multi-well platesusing devices with multiple pins.27
Alternatively, the gap can be created by physically excluding the cells using inserts. Suchinserts create a linear or circular gap by adhering to the treated dish bottom andpreventing cell growth in a predefined region. A linear exclusion commercial insert(www.ibidi.com)is shown in Figure 2 (examples ofother insert suppliers include Cell Biolabs, and Platypus Technologies). The wound-healingassay is initiated by removing the insert. The advantages of using inserts include requiringfewer cells when seeding as well as more reproducible gap sizes compared to the scratchmethod. While the gap size created by the insert can be more reproducible, cells clinging tothe insert can be torn out of the monolayer leaving jagged edges. This means that the gapedges are not necessarily better defined compared to a scratch assay. In addition,commercial inserts are more costly than using the scratch method. Inserts can be re-used tomitigate the cost, but over time their adhesion to the bottom of the dish may becompromised.
If the graph is plotted with area in μm2 and time in hours, thenvmigration conveniently can be expressed in units of μm/hour. Figure 6 illustrates four timepoints (A-D) from a scratch wound healing assay carried out using HUVEC cells with thearea of the gap plotted over time in Figure 6E. Note that the graph of area over time is linearfor the early time points; however, at later time points where the gap is nearly closed,the image analysis routine fails to detect and measure accurate gap areas as shown inFigure 6E. For this reason wedid not fit the curve right to the gap closure. In fact, as the trendline is approximatelylinear, it is sufficient to end the experiment and fit the data up to the half-closuretime. For the experiment shown in Figure 6, the calculated t1/2gap is3.27 hours and the cell sheet migration rate is 8.35 μm/h. While thet1/2gap measurement can only be used to compare wound healing assayexperiments for which the initial gap is the exact same width, the cell migration rate ismore broadly comparable across datasets with varying wound areas, and is probably the bestmetric for quantifying the wound healing assay.
Designed by Numark in collaboration with Serato, NS7 represents the pinnacle of DJ performance control. NS7 combines an all-metal chassis with adjustable torque, motorized, aluminum-turntable platters, vinyl, and a professional audio interface to deliver a complete performance solution that will satisfy even the most hardcore turntablist.
A full complement of premium faders, knobs, buttons and our new Strip Search (patent pending) virtual-needle-drop control integrates seamlessly with the included Serato ITCH software. NS7 delivers hands-on control of every aspect of Serato ITCH and sends data to the software more than ten times faster than standard MIDI resolution, resulting in a rock-solid hardware/software DJ system that works with Mac or PC and feels as good as it looks.
It could be possible go a bit further improving the recording cappabilities of mixtape. Adding turntable vector recording (check the scratch track automation recording) or if you prefer doing it with SSL, split the turntable output and recording the serato vinyl directly alongside SSL audio from deck. Routing it properly gives you the chance of editing even scratching mistakes. Not clever solution due to lack of imagination in most DVS developers and the mantram about realdjing everytime a feature makes things easier. It could be true multritrack and gesture recording.
I have a major problem with getting my Serato SL1 box to work with Serato DJ. i have a new solid state hard drive and i lost all of my files. I dont have the original SL1 driver, and cannot find it, ive looked everywhere. What do i need to? my sl1 serato scratch live box is not working with the serato dj. thanks djs
I have a laptop running win 8.1 and i considering buying an SL1 and using it with scratch live. Why? because i am only doing this as a hobby (not making money or leaving my basement) and i want the cheapest way to a functional serato system. Is this crazy talk in mid 2018?
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This Easy Homemade Brownies Recipe makes cakey brownies that taste like box mix brownies! With perfect crisp crackly tops and chewy centers, these made from scratch brownies hit all the right notes.
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The examinations are closed book. You are not allowed to access notes, books, reference materials, or electronic devices at any time during the examination or during breaks. Unauthorized access to notes, books, reference materials or electronic devices may result in your test results being nullified by the IRS. You will be provided scratch paper and pencils at the test center and you are not allowed to leave the testing room with notes taken during the examination.
Bartonella henselae bacteria cause cat scratch disease. They live in infected cats' saliva (spit), but don't make the animals sick. In fact, kittens or cats may carry the bacteria for months. Fleas spread the bacteria between cats.
The first sign of a cat scratch infection is a blister or a small bump several days after the scratch or bite. It may look like a bug bite. Within a couple of weeks of a scratch or bite, one or more document.write(def_lymphnodes_T); lymph nodesclose to this area will swell and become tender.
Doctors usually diagnose cat scratch disease based on an exam and asking whether the child was around a cat or kitten. During the exam, the doctor will look for signs of a cat scratch or bite and swollen lymph nodes.
Most cases of cat scratch disease do not need any special treatment. Doctors sometimes use antibiotics to treat a severe case. If your doctor prescribed antibiotics, give them to your child on schedule and for as many days as prescribed.
Kids with cat scratch disease don't need to be kept apart from other family members. Let your child rest as needed. If your child feels like playing, encourage quiet play while being careful to avoid injuring swollen lymph nodes. To ease soreness, you can give your child acetaminophen or ibuprofen.
Cat scratch disease is not contagious from person to person. The bacteria spread by the scratch or bite of an infected animal, most often a kitten. They also can spread if the animal's saliva (spit) comes in contact with a person's eye or through broken skin. Sometimes more than one case happens in the same family, usually through contact with the same infected animal. 2ff7e9595c
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