• Safe and secure

  • Quick and easy

  • web-based solution

  • 24/7 Customer Service

Rate form

4.0 Statisfied

965 votes

The Instruction of Finishing Determination Descent Form on the Internet

Get and personalize the perfect Determination Descent Form in the CocoSign template library to fulfill your needs and save your cost. If you are still wondering how to fill out Determination Descent Form , you can check out the below tips to start.

Discover the signing area

Write your signature

Click "done" to save the form

  1. In the beginning, you should find the right form and open it.
  2. Next, take a look at the form and know the required data.
  3. Then, you can begin to fill in the details in the blank form.
  4. Fill up the check box if you are eligible to the condition.
  5. Take a look at the form once you finish it.
  6. Insert your esignature at the bottom.
  7. Select the "Done" button to save the document.
  8. Download the form in PDF.
  9. Chat to the support team to get more details to your confusions.

Choose CocoSign to simplify your workflow by filling in Determination Descent Form and adding your esignature shortly with a well-written template.

Thousands of companies love CocoSign

Create this form in 5 minutes or less
Fill & Sign the Form

CocoSign's Guide About Finishing Determination Descent Form

youtube video

How Do You Get Determination Descent Form and Sign It Online?

hi everybody thanks for joining today.this is Jason key at Harvard Medical.School associate director of sp grid.hello everybody it's a real pleasure.today to have Shore sheriff's on our our.webinar so thank everyone it's excited.to hear about new features coming and.rely on through no Shores leads a group.at the MRC and the UK the developer of a.lot rely on ditch I think as Corey M.tools goes is certainly one of the most.popular and has probably sold more in.video cards than any piece of software.that I'm aware of.so sewers are you there yes sounds good.go ahead cool yeah thanks Jason thanks.for the invite to speak for your for.your audience so today I'd like to.discuss what's what's new in the orange.version of reliance three point naught.and the first thing that's new is dead.words until now had been mainly.programming on the rely on by now on my.own with with kind of a limited part of.the code being developed by shouta he is.a student PhD student who developed.helical reconstruction now I had two.postdocs who are helping me a lot with.developments in site.reliance core and and everything around.it so and I will present work from both.of them today so there's just anchors.even off who's developed Bayesian.polishing and the CDF refinement and the.bean field estimation I'll speak about.today and then takanori Makani he's been.involved with the implementation and.testing of multi body refinements has.done lots of tweets throughout the.software also to helping make all our.our kind of body of software around with.work with the new implementations from.jusenkyo and has been extremely helpful.in in providing user support and many of.you who have seen all those answers on.ccpm email list and then lead on is a.new page c stand in my group who's.working on.mission of class selection in first.instance and her burg hasn't left yet -.incorporation in rely on three but.hopefully it will have become part of.future versions so I will also not speak.about today about work done by my.collaborator in Sweden Erik Lindahl who.was in Stockholm and they did the GPU.acceleration of rely on - and now as.part of rely on three they've done the.if done similar unwrapping of the of the.very intricate loops in reliance inner.core such that you can use vectorization.on CPU so now thinks real and three can.run on on multi-core CPU clusters much.faster than before about five to ten.times faster and thereby it becomes I.think in cost efficiency is kind of.similar to the professional type of.double precision Nvidia cards.whereas I think single precision gaming.type of cards are still a lot cheaper.than running on the CPU okay I thought I.just go from the from the top to the.bottom in the in the kind of job type.window here on the left starting and.highlighting and what is new though one.thing that is new is that Takanori has.made a a C++ kind of reverse engineered.version of motion core to people on.Twitter have been calling in to and rely.on core to on the on the GUI it's called.reliance or implementation so we did.this originally to be able to take.forward more metadata involved in stuff.frame alignment Daisy and polishing.although in the end it doesn't seem to.matter very much so what distinguishes.it from a it's it's what it tries to.replicate motion core to is that this is.CPU only it's multi-threaded and.provided you have a.a 12 or a 16 core machine and then you.can use integers times of the number of.frames that you have for the number of.threats which that each threat has as an.integer number of frames to process then.speeds are our batter or equal to them.then it's GPU processing on indices that.version as I said the original argument.of doing this was passing around of.metadata one difference as well is that.this is open source and three for all so.we there's even if you're a company you.can you can use this and overall I rely.on is all open source and we take a.point in that in that open source.software even though people can copy.pieces and put it into your own and so.on that you could perhaps perceive as a.threat to your own software ultimately.that is impact that your development.make and a field grows faster if people.use open source which other people can.build on and I will show you examples.also today of things that we've taken.from other open source packages and.incorporated into real arm so in the.automated particle picking job type one.thing that is new you can now provide.the 3d reference and this was a popular.request and we uses in our fully.automated on the fly and processing.scripts that I will mention later on.where you take a 3d map you projected in.with it really give an angular interval.into all different orientations and then.you use those for reference base picking.another new thing is this laplacian of.gaussian then picker and that's there's.a there's a second tab here at the.platen tab and this is meant to be.relaxed for the moment approach to do.reference free auto picking it's based.on a laplacian of gaussian filter which.is sensitive to blobs of a given size.and then you can you can vary this from.a minimum to the maximum size so if you.have more or less spherical particles.these.maximum size can be very similar or if.you're very elongated particles then you.can reflect is by a very different their.values of the minimum and the maximum.diameter the the filter itself is we've.trying to normalize the threshold such.that just default values give reasonable.pics if not perfect pics by far but.enough to get you started again we'll.see this later on in the completely.automated way of processing your data we.aim to get thresholds that are valid for.a whole wide range of different types of.data set now the way the program works.internally it will search the.micrographs for sizes of blobs that are.way smaller then you've then you've.specified from your minimum to your.maximum diameter it will do the sizes of.the minimum the average of the two and.the maximum size those are your that's.your expected size range and it will.also search for blobs that are way.bigger than your accepted particles and.the too small and the two large searches.serve to identify regions of the.micrograph where we think your particles.are not there so these are trying trying.to find the true negative areas and.focus your searches on on areas where.your particles are at of the right size.this is a very favorite favourable test.case perhaps just ribosome data sets on.a direct electron detectors and contrast.is very beautiful you can see most of.the ribosomes are picked there's some.false positives here which result from.from a compromise that at some point.there's just one general vessel to pick.them all all kind of different quality.data sets so the idea is that at least.for a wide range of possible quality.datasets you get at least a subset of.particles that is good enough to then.start your on-the-fly processing.approach which I will discuss later so I.personally wouldn't rely.just log picked particles for your final.data processing but it might be a good.alternative to what used to be normal.and rely on that you first pick several.micrograms over a few dozen of them by.hand and then use a reference free to.the class average in to generate.references now you can kind of start.from lock picked micrographs and go all.the way without any user having to pick.particles by hand then on the 2d.classification job type there is a new.option to use fast subsets and this is.particularly useful for large data sets.and this is an idea that we got from.from Niko with oreos and Tim grants.paper on system which is published in.any life and this is to use fewer.particles in the early informations.because if you have picked a million.particles from your from your two days.worth of data collection and then 2d.classification will assign random.orientations to to all of the 1 million.particles normally and calculates.spherical like blobby references and.then during the first few iterations you.pass over each iteration a million.particles and these circular references.become slightly less circular in the.first iteration and them slowly converge.towards in the the hopefully.representative views in your data cells.but in practice you don't need to pass.over a million particles in those first.few iterations because the references.are still very low resolution so you can.you can get away with much fewer.particles so we used some heuristics.there it's it's hard-coded inside the.call it's similar to what Nikko does and.I personally probably wouldn't use it if.your data set is no bigger than say a.hundred thousand particles then the.differences aren't that big.but if you go beyond that then the data.processing will become a lot faster and.what we've seen the results are of.course different but they're not.qualitatively worse than just passing.overall data then in.a model generation in 3d already in rely.on 2.1 we implemented stochastic.gradient descent and an idea originally.proposed by a leapin Johnny and Marcus.blue acre in Toronto and one of the.bases of the cryo spark package so in.2.1 our implementation was sub-optimal I.would say so it would work if the data.was very good but it would cry a spark.definitely worked a lot better in it for.many data sets than the rely on initial.model generation though in round three.because Christ part 1 was still open.source so we could actually have a look.around and we actually implemented.something much closer to the private.spark algorithm with most of the.parameters those who've used trying.spark will recognize them now so you can.you can do things very similar in the.real aisle and we've seen that that.actually has made the gap between krile.spark and rely on the initial model.generation a lot smaller if not if not.absent in many many cases so initial 3d.mobile generation within rely on de novo.has become a lot more robust than it was.in in previous versions you can now also.refine multiple initial models at the.same time etc so that's inspired by cryo.spark now then one of the big changes in.rely on 3 is the inclusion of multi body.refinement and multi body refinement as.I said was was co-developed by Takanori.in my lab and it is basically it builds.upon the the concept of focus refinement.with partial signal subtraction which.was already available in in rely on too.and the idea here is that you have a.part of the protein complex which you.are interested in to refine separately.from the rest of the complex for example.because it is flopping around all.because some nitrogen 80 other types of.heterogeneity happening there now in.schematics here is then a part.interested in in red and the part we're.not so interested in in yellow this is.gamma secretase will be first it is back.in 2015 the experimental images are of.course a projection of both the red and.the yellow part and then there is no.he's on top now already on ribosomes and.so on back in 2013 and 14 we would do.focus refinements wherever we put masks.on one subunit and then at every.iteration in order to focus refinement.on that subunit and be kind of.insensitive to changes happening in the.other subunit.the disadvantage of that is that the.mast reference projection for example if.the mask words can say only the red.parts then the reference projection only.have signaled the red part of the.protein but the the actual experimental.image has noise but it has the red part.and the yellow part of the of your.complex so the comparisons between the.experimental image and reference.projections for mast refinements alone.are inconsistent the yellow part is.missing but the idea there was to then.subtract to make a mask around the.yellow part and subtract in silico the.reference projections from the yellow.part from the experimental image thereby.generating a new version of the.experimental image which only has.projected density hopefully the this.abstraction were perfect of the of the.rap part that we're interested in and.then the focus refinement thereby.becomes the comparisons with the mask.projections become consistent again now.that was then used by Callaghan Ian here.in Kiev in a guy's lap on the.spliceosome with great success and I.think many people have used bogus.classification focused refinements and.to to great success now multi body.refinement it goes one step further and.it provides a iterative partial.subtraction so you don't just do this.once at the beginning of a refinement.but you do this iteratively at every.iteration usage.correct and you now have not just one.part of the structure that you focus you.only find the Dom but you can divide.your complex into as many rigid bodies.as you would like to refine separately.and the multi body refinement will do.the subtraction during every iteration.iterative I'll come back to that for an.example of the 3d and floppy protein.others here so imagine we have three.three domain protein and the M domain.and the our domain and the C domain are.connected by flexible linkers such as.you have independent motion of M with.respect to R and C with respect to R the.way multi-body.refinement works is just like focus.refinement on the M domain we're going.to make reference projections of the RC.domain and subtract them from.experimental image which has an RNC.domains which leaves a experimental.image where the M domain is still.present in the R and the C domain are.subtracted and then we do a refinement.of this image with the reference.projection of only the and the main now.that will give a new set of orientations.for the M domain which hopefully is.better than the initial guess that we.got from our consensus refinement where.we just imagine that this was once a.rigid structure and we refined all the.images just by projection matching.against it now you can already see here.these the partial signal subtraction.wasn't perfect and that is because the.orientations of the rnc domain perhaps.we're not perfect either so the M domain.has some problems in its orientation but.also the RNC and that depends on how the.different domains flop around in this.particular instance of the experiments.of particle so we do this for the M.domain in the first iteration but.simultaneously we also do this for the.arthelene so we subtract the M and C.domain and do a focus refinement then on.the our domain that gives also new.orientations for the our domain and.likewise also for the seedling in the.next iteration the subtraction of the RC.signal for the focus refinement of the.EM and all the other ways as well.becomes better because these angles are.better so you iteratively do better.partial signal subtraction and while you.accumulate now three sets of.orientations for each experimental.particle which express the relative.orientations of the and the AR and the C.domain with respect to the overall.consensus orientations from the first.refinement and this iterate through all.the way until the orientations don't.change anymore and there are the.resolutions of the separate.reconstruction for the MDR and the C.domains don't improve any longer that's.and that way it's similar to just alter.we find it so the example I will show.you is from from another splices and.complex the B complex which is also done.in the guy group and the the Takanori.made masks around four different domains.so the core domain here in the middle.was the highest resolution in the.consensus structure you can see quite.nice structure the foot domain was still.okay as well and then the helicase.domain here already was getting a lot.more fuzzy you can see density here and.for the SF 3b domain on the top very.poor and then this Russell would almost.completely disappear so there is.partially dependent partially.independent rotations and translations.of these four domains with respect to.whether now the consensus map and that's.here shown in the in the top panel it.shows different slices along along.desert axis through the consensus map so.this is the core domain here which is.very nicely defined and then the foot is.down here so I'm a bit lower in that.here you can see the foot gets a bit.fuzzy but it's also still okay and then.the SF 3b and the helicase domain this.is the helicase and this is f SF 3b are.a lot worse so they are very very fuzzy.in the consensus structure now after a.multi body refinement doing this with.four bodies at the same time you now get.for supper.three constructions of the individual.bodies and the core improves a little.bit but it was already quite good the.food domain you see this was quite fuzzy.here this was a bit fuzzy it gains in in.resolution although already it was also.quite good and then and the most.dramatic differences are seen for the.helicase domain especially here at the.outside and for the es f3v domain so.that's also then reflected if we do the.local resolution estimation on the.consensus structure local resolutions.beyond 10 angstroms here at the top and.if I now place the individual bodies of.the four body refinement on top of each.other that's a problem we don't really.know what happens in the interfaces but.I'm just rigid partly docking them into.the consensus structure here then you.can see that the local resolution is.improved throughout the structure and.the same as mechanically by D F a.seekers before and after multi body.refinement now one interesting aspect of.multi body refinement is that you get.the set of orientations for each of the.bodies for every experimental particle.so every experimental particle now has.four sets of orientations and you can do.a principal component analysis on those.orientations to find out what are the.most common type of motions that happen.in the in the entire set of particles.and then you can just look at how much.variance each of those emotions.correspond to and here the first motion.as most of the variance the second.motion was the one that the negai group.thought was most interesting and you can.then make movies along this type of.motion to get insights on how the the.molecular complex what kind of emotions.are present and you can see for this.place is owned or the spliceosome that.the SF 3b and the helicase domain kind.of have this concert of motion with it.with each other on top of the core where.as changes here in the a lot smaller.which is kind of which which agrees with.the resolution of the fruit originally.being quite and quite good already coke.I think this is one of the main.attributes of the multi body refinement.compared to the partial signal.subtraction and the focus refinement is.you do everything at once and that does.allow you to get overall motions out.with these kind of movies now another.major improvement in rely on three.compared to rely on previous versions of.real um is the way we do Department for.particle based motion correction which.traditionally was called particle.polishing in reliable and because the.Jarett that's because it generates shiny.particles when signal to noise ratio is.improved.now Bayesian polishing has its name this.was done by just thank you Bayesian.polishing has its name suggests uses a.regular eyes likelihood function to.follow the per particle motion tracks.through the movies of your of your a.direct electron detectors now the light.load function is is just defined by by.the movie frames and the the the prior.that you need for the recognized light.hilt function or the regularization.terms are based on smoothness in.velocity so you can't have very big.accelerations or the accelerations on.the acceleration and then also on the on.the observation that neighboring.particles tend to move in similar.directions so we we impose this with a.Gaussian prior on keeping the tracks of.neighboring particles the same and then.just anko's algorithm looks at for each.particle for each micrographs it looks.at all particles simultaneously and.movie frames simultaneously to find.these most likely track given the.regularization terms and the output of.that is then going to all these PDF log.files where you can see.these kind of plots and we present you.know in the paper on this which will.soon be published in International Union.of crystallography journal we use.Plasmodium ribosome beta galactosidase.and gamma secretase the tracts alone are.kind of interesting to look at that they.don't really tell you much what's going.on but one thing that's that's.interesting is that if you look at the.fall-off of the signal with spatial.frequency then for each of the movie.friends and already in row in previous.versions of rely on we model this little.B factor and B factors being more.negative here and means there is less.high spatial frequency content in the.images than the old polishing method for.the Plasmodium ribosome captured the.initial motion very poorly which led to.always the first few two or three frames.having relatively poor B factors and in.the new polishing and these tracks.initially are followed much more.precisely leading to much higher initial.B factors for the first few frames and.that's where radiation damage is still.the lowest those are your most important.frames so by by rescuing these initial.frames you can get more out of your data.and that's I guess then the main.contribution of the Bayesian polishing.so that then leads to resolution.improvements consistently sorry about.that.and that then leads to resolution.improvements for all the cases that that.we've run this on and another way of.looking at it is that you can look at.these B factor plots where you you plot.the logarithm of the number of particles.versus one over the resolution squared.and you can then see that the Bayesian.polishing gives you better be factors or.in other words you can get to the same.resolution and for these data sets it.turned out more or less processed 60%.fewer particles you can get the kind of.same resolution now the the second.development by Janko is this other job.type CTF refinement and the CTF.refinement actually includes two.different types of calculations one is a.refinement of the CTF parameters.basically D focuses per particle is.there is the most important thing here.and the other development is estimation.of how much beam tilt is present in your.data so that beam tilt leads to the coma.if you haven't done your coma free will.alignment of the scope while then you.have been filled in your data which.leads to phase differences in the images.which you can estimate and correct for.so if you want to do Perkins estimation.then that allows one that includes one.more parameter for each particle so if.you have a hundred thousand particles.that gives you a hundred thousand.parameters that's enough to overfit your.data so you have to do this using this.gold standard separation of two hot once.you go get overfitting of your data this.is just a technicality but which is.implemented in real um and you don't.need to worry about it but you would if.you would like to do this in other.pieces of software perhaps now the.reason why we think we can do better.than just CTF fine for or or GCTF in the.beginning where you estimate the focus.from power spectra of micrographs is.that the power spectrum this is kind of.the the power spectrum signal so you see.the rings but they lie on top of quite a.bit of background noise and here the.dotted grey lines kind of indicate how.much noise there is on this signal.whereas if you do the per particle.comparison with the reference projection.of what the signal is in the particle.images and compare that to the reference.then you can see both the positive and.the neck.modulations of T of the CTF you can see.the zero crossings and importantly the.amount of noise on this type of signal.compared to the amount of signal that.there is is much better so you can.estimate your the foci and more.accurately if you do reference based if.you know what the signal is basically so.we tested this and this is not.necessarily recommended way of running.this but just to show you how how far.this can go we tested this on the data.sets I guess is from Dmitri eunuch ISM.on the hemagglutinin which was tilted at.40 degrees to overcome preferred.orientation so we just started from the.purp micrograph G C T FD focus.estimation which is pretty badly off for.for the parts that are much closer and.much further away from focus due to the.40 degree tilt of course now originally.I think the me to publish this data set.at four point two angstroms resolution.and then the other Dmitri take you know.it process these data and warp and then.use cryo spot to get to three point two.angstrom resolution so when we use real.arm three for this we actually had to.use four cycles of CTF refinement.because the initial reconstructions from.the average the foci of the entire.micrographs only goes to about seven.angstroms resolution and if you then.refine the focus you suddenly make a.jump to too close to 4 angstroms.resolution so that goes from the gray to.the orange line but the once you have.that reference you now have a much more.detailed reference you can then again do.better and D focus refinement so that's.why in if we do this four times then.things start to stop improve and then we.do the Bayesian polishing we classify a.bit and do another three runs of the.focus then in the end we end up with a.three point one angstrom structure and.the the kind of pitches that we see in.the map as we go along this process the.kind of improvements and so does a fic.calculations against atomic model.if you then look at the refinery foci.for particles within one micrograph you.can see beautifully this filtered aspect.of the micrograph back in the D focus.values and they're probably few outliers.where refinement has gone off but.overall you can rescue these at the foci.which kind of illustrates that you can.you can you can correct the foci values.which are here in in quite a substantial.amount of quite a substantial range of.five thousand angstroms now VIN tilt.estimation rely on since version one.point four two point zero I think had.the option to correct for beam tilt but.we never knew how much being tilted was.present in our data and it was jusenkyo.who developed a method to estimate the.amount of being tilt from again.comparisons of the individual particle.images with clean reference projections.and see if there's systematic deviations.in the face errors within between those.images now one thing that this allows us.now to do is is to collect images using.the beam shift rather than stage shift.so what we typically do is refocus.somewhere on the carbon and then we take.a picture in the middle of the whole let.me remove this stage to the next hole.and repeat now after you move this stage.you have to wait for drift to settle so.this becomes a rate limiting step so.therefore it's quite popular to use beam.shift to help to collect poor images in.each hole and then move to the next hole.now if you'd use beam shift in the.microscope to to shift your beam that.actually introduces a bit of being tilt.now the amount of beam tilts you get.from shifting within a hole is not a.problem.but what you can do what would be.potentially even faster and this is.together with when harkened at the MBL.and Heidelberg is to take images in nine.holes for images in each hole for.example and then move the stage to the.next position way in the middle off of.another nine holes so then by there.there.for again increasing speed of a data.acquisition because you don't need to.shift the beam and rim was developing.methods using the optics of the of the.microscope to correct for the actual.beam tilt that shifting the beam so far.out induces so you can use the.microscope hardware to take images here.that do not have beam tilt in them but.as a control we've also collected data.where he just ignored the tilt and just.took the images using the beam shift.anyway so we thought that would be a.nice data set to test our method of.estimating and correcting for beam tilt.on so these are on April ferritin he.took actually five images for who and.then used nine holes one dataset where.he uses active beam self compensation.that says Hardware method to correct for.being tilt and then his controlled.dataset where he didn't do this now.his active beam tilt compensation works.very well and you didn't get a 2.1.angstrom or so structure out of these.400 images of april ferritin now if you.don't use any correction for the beam.tilt nor hardware or software then you.get a much lower resolution which is i.think it's about 2.8 or 2.7 answers now.by estimating the amount of beam tilt.from the particle images and then.correcting in rely on for it.we get we get back to more or less the.same the reconstruction perhaps even.slightly better than what we've got.we've got in his hard way correction and.this is just to show you this is for the.nine hole so this is each for each stage.position you take an image in the.central hole which will have very little.beam tilt and therefore the the.systematic differences in in phases.between the Fourier components that's.what we're looking at here at that.negative blue and positive red and.yellow.are more or less absent when you take.images down to the central hole but if.you shift the beam upwards all the way.to the next hole then you introduce.these face difference in the direction.perpendicular to that shift so you get.the positive phases on one side of the.spectrum and negatives changes phases on.the other side and when fits.no just Cinco fits functions but this.goes to the cube with resolution we know.how they go so the sample can fit these.functions through them and thereby.estimate very actual what the amount of.beam tilt that you get for each of these.holes and the direction of the beam till.- and then the correction for that leads.to these huge improvements in resolution.now one other thing that's also new in.round 3 is yacht sphere correction and.this at the moment at 300 kV only.matters for really large particles.probably mostly icosahedral viruses it.will become more important and Richard.Henderson Christmas who are now trying.to push for lower resolution 100 KB.microscopes perhaps with effect then.these things will become Department more.important they also become more.important with higher Rd focus but if.you do have a large virus and this is an.example that we show P 22 virus which is.about 700 angstroms across if you do use.evil sphere correction according to the.algorithm that was proposed by Chris.Russo and Richard Henderson earlier this.year then you can see from not doing any.evil - in correction you can improve on.resolution using keynote speaker ection.after say this is not implemented inside.rely on refine this is only part of rely.on reconstruct so after you've done your.refinement you need to take the data.that star file and do reconstruction for.each half of the data using evils for.your correction because you don't really.know which is the correct hands you have.to do this for both one you could have.flipped your data around an old number.of times doing image processing so in.practice you need to try both hands in.the evil - a correction and one will.become better the green line and one.will become worse which is the orange.line then I mentioned.in the beginning that we've now worked.on on scripts to do on-the-fly.processing without any user interaction.we call this rely on it still pi because.you just need to rely on it there's no.more space for you to interfere with you.with your data processing and this is.just a screenshot it's a it's a script.written in Python and it will schedule.and execute jobs just within the normal.rely on GUI pipeline so it's it's cut if.you like it's a Python well it's.basically a command-line interface into.that what you normally would would deal.with in the GUI and that allows you to.write scripts around it and then we use.Python but you could have used a bash.script as well and we hope that these.Python scripts we provide two one for.on-the-fly processing called Orellana.pi and want to make beef actual plots.which I showed before which is the.logarithm of the number of particles.versus one of the resolution squared.after refinements so we hope that these.scripts will will be an inspiration for.others to to write their own kind of.semi automated procedures so to rely on.the PI script will iteratively import in.micrograph movies it will do the motion.correction CTF estimation do this.lock-picker and then go from extracted.particles straight into 3d SGD from.their own into 3d classification use the.best class to do reference based they're.also picking and then get a final kind.of set of particles which you can then.use for 3d classification or refinement.and to test this we used gamma secretase.which was originally collected here by.search and by postdoc in my group with.an a UT Southwestern and we chose this.dataset because this is not your kind of.bog-standard test dataset like april.ferritin or vita galactosidase this is a.130 kilonewtons ordered mass membrane.protein complex so this isn't definitely.not very easy data and we then compared.how xiaochun's very expert selection of.particles which gives a 3.2 angstrom map.in real arm 3 it gave a 3.4 ounce remap.and rely on 1.3 budget.tchau gendered so this the improvement.- green is due to Bayesian polishing and.CTF refinement etc but we then this.involved Xiao Qian looking at six.subsets of the data do individual -.declassification select all the nice.classes etc how that would compare with.a fully automated processing by the.script is then given by the orange line.so that gives the 3.3 angstrom structure.so fully automated processing of gamma.secretase in rely on 3 is slightly.better than rely on 1.3 but it's not as.good as Xiao Chen would have done in.reliant 3 so but the difference is.getting smaller and we will keep working.on this script this is definitely a work.in in progress and I think not too far.in the future you'll see that you can do.all the way up to kind of consensus.refinement will probably be come.relatively easy to do in a fully.automated manner for many many data sets.I just wanted to finish with two data.sets just to show what rely on 3 can can.improve compared to what it can do what.you can achieve with it one is this.Empire entry by three realms of.Romanians group their own beta.galactosidase where they initially.published in 2.2 angstrom map and then a.map the big that they said was Thomas.resolution so the Bayesian polishing.gives you these kind of B factor so.again whereas previously Bayesian.polishing would would start at minus 200.here so now the first few frames have a.lot more high resolution information.contents after dealing well with the.with the initial motion of the particles.which is very fast surprisingly to us.this data has a significant amount of.beam total is about point to Miller rats.in the X direction and you can see that.here in this in this face make so phase.difference plot the Duchess n curves.program to rights out so there's neck.systematic negative differences on one.side and positive differences on the.other side which is the clear sign of.beam tail so this is one of the reasons.why we could.on resolution program used per particle.astigmatism in this atomic resolution of.paper and we think that that might have.been partially describing the effects of.being tilted now this is how our.processing scheme start we start with a.large set of auto pay particles we do.the focus refinement and tilt refine.that if you do just one of them either.the focus refinement or tilt we find you.go from the gray of a seeker today to.the dotted or the solid line orange one.if you do both of them at the same time.that their effects are cumulative you go.to the duty to the pink line here then.the effect of Bayesian polishing on this.data set is quite large as well you go.to the purple line then we tried a.little bit of classification and undo.another of the focus until division.improvements were a bit smaller and you.can see throughout this process that the.map actually gets better and and.features start to appear that should be.there so this final green FSE curve.corresponds to a 1.9 angstrom resolution.map where we see beautiful rings.beautiful holes insects 6 membered rings.but not yet that much in in five men.between switches as expected at 1.9.angstroms resolution I told you we had.this this Python script to calculate.this B factor plot which makes.automatically for you subsets of the.data set and does refinements for each.of the subsets and then it looks at the.post process 1 over resolution Square.and makes this nice PDF plot for you and.from that we estimate now that the B.factor for this data set is 56 squared.angstroms and that compares with 91.squared angstroms in rely on - so you.knock off more than 30 squared angstroms.from the B factor going from Milan - to.reliance me due to a combined effect of.the focus refinement beam tilt.correction and Bayesian polishing and.locking 30 squared angstroms often rely.on sued and improves resolution from 2.2.and run to 2.in 1.9 in rely on three and that's.that's on this plot so the orange.version of.alarm gives the orange line and the.green version land to gives the green.FSE curve this is the FOC of three rams.model versus our map so we didn't do any.attempted refining Sherman's model we.just took that at face value and we see.if we the asked the reported resolution.of 2.2 to 1.9 is reflected in an.improvement in the FOC of model versus.map which is indeed the case.interestingly Sri Rams map which he.claimed was an atomic resolution is.correlated with his own model up to the.to the gray Kerr.so the Reliant 3 map is better than the.atomic resolution map from the recent.structure paper although I think calling.at atomic resolution is probably not.correct yet so april ferritin is then.the last case I thought I'd mentioned.this is another data set by Byron Hagen.collected at the NBL taking now seven.images per hole not doing the beam shift.all the way to other holes and is just.conventional stage shift I think it's a.bit all our dataset so he collected 1200.movies at a point eight angstroms per.pixel just to see he thought he'd see.how well his crimes would do we to.pre-process these data using the rely on.little PI script again the initial set.of particles redundant through.declassification selected nice classes.which you see TF refinement and Bayesian.polishing on there was hardly any tried.being too estimation that there wasn't.hardly any being tilt in this data said.because I think min aligns his.microscope very well and that then led.to a reconstruction through one point.six five angstroms resolution so this is.very close to two times this number I.think we're only two or three shells.away from the Nyquist frequency I do.think this resolution estimate is true.do we see beautiful rings in in proteins.we can see even bigger blocks for sofas.here and overall I think density looks.and pretty good reflect.then as you would expect for inspire.angstrom resolution map that was all I.have to say you can download reliant 3.as long as its beta we have this.download on bitbucket.and and will hopefully soon go stable.and then we'll move everything over to.the yet half as well so thank you for.your attention.these are all the people involved Jake I.mentioned or Eric and Takanori and Jason.ku and Lee Eric Buren and Ari are the.people in Sweden and Jake and Toby.maintain our computing infrastructure I.am very grateful for all users who.provide us with very insightful and.detailed but reports that just don't say.it doesn't work but actually tell us.what goes wrong and I thank you very.much for your attention that was great.thanks there's just been a couple of.questions in the chat if I can pass.those over to you yes though we have one.from Long fate which is how to use multi.body refinement on proteins with high.symmetry that's a good question if if.you have a high symmetry and you want to.maintain that symmetry you're imposing.that you that all the different bodies.they they will they have some motion.with respect to each other otherwise you.wouldn't do multipotent refinement so if.you want to still refine multi body in a.high point group symmetry than you then.your basic assumption is that all the.motions still obey that point group.symmetry which may very well not be true.so if you have a kind of typical example.of an icosahedral virus from which.floppy domains are sticking out of than.those the floppiness of those domains.probably do not obey icosahedral.symmetry and then rather than using.multi body refinement you should use.something which the program called rely.on particle symmetry expand and this is.this procedure of symmetry expansion is.described in 26.in review in methods in enzymology the.book that was edited by Tony Crowder.which is all about classification of.images in in rely on and that describes.on how you can use you can expand your.datasets which would have 64 which would.become 60 times larger in the case of.64th icosahedral symmetry you can then.use c1 refinements on these expanded.data to deal with that kind of.sloppiness very short probably you.wouldn't use multi-body on on very high.symmetry groups last question.I'm trying something smaller with the.virus and I I want to know how I could.use particles of subtraction on a.symmetry expanded stack of refined virus.particles or should I just go back to.using or refining the subtracted.particles in c1 yeah so you can use.symmetry expansions first so you'll get.a much larger star file with where each.particle is replicated and then you do a.subtraction using that expanded star.file so now each original particle will.actually become image will become.multiple images on desk.each where a different part of the virus.is still there the signal and then you.can refine all that in NC 1 or.classified NC 1 there is also a new.option on the particle subtraction job.type which it allows you to provide the.center of rotation from which you would.like to do the particle subtraction and.that might allow you to then use shorter.smaller box sizes of on your subtracted.particles although that will require a.little bit of scripting around and some.of the command-line programs that's not.only that's not possibly only through.GUI.okay another related question on.symmetry is what tools would be useful.in Milan to deal with protein assembly.that has a highly symmetrical region.while other parts of asymmetric you.there is this localized masking approach.that Xiao Tao also wrote and it's it's.you can find it on the GUI on the on the.on the weekly sorry on the wiki page of.rely on the left side there is something.just under the helical reconstruction.there's something on localized masking.where you can kind of impose local.symmetry during refinement in rely in.practice we have not encountered many.cases where it actually yields a big.improvements perhaps that's why we never.published it so in practice we just.often just refine in in c1 and you can.then still average over the over the.symmetrical copies afterwards to make.them better but inside the iterative.refinement it doesn't really help often.but it might be that your case is the.first okay there's just a couple more.questions and then I think I'll let you.go.so somebody calls SPL would like to know.how well order does the multi domain.protein need to be for multi domain.environment to work oh it doesn't.necessarily need to be very ordered as.you could see the splices and parts of.it were in very ordered at all I think.the main limitation in application of.multi body refinement is this the.minimum size of the individual bodies.because you're going to subtract all of.the rest away there needs to be at least.I would say about 150 kilo daltons worth.of mask within each body if that's the.case then quite big motions can be can.be refined perhaps if you have a.completely floppy beat of you know a.string of five beats and there's very.large amount of.motion between each bead such that the.whole consensus refinement would but.surely fail then that would become a.limit to but rather than that it's.mostly in most cases it's as the minimum.size of the individual bodies which is.much more limited than the entire amount.of motion if you if you get a reasonable.consensus refinement then and the.individual bodies are large enough then.multi body refinement could work so I.guess thank you very much for the great.seminar that was really useful it's.certainly something we'll be around here.I don't suppose anyone else in the room.has any questions then okay so thank you.very much okay thank you good bye done.good bye.

How to generate an electronic signature for the Determination Descent Form online

CocoSign is a browser based software and can be used on any device with an internet connection. CocoSign has provided its customers with the most useful method to e-sign their Determination Descent Form .

It offers an all in one package including safety, low cost and easiness. Follow these tips to add a signature to a form online:

  1. Ensure you have a efficient internet connection.
  2. Click the document which needs to be electronically signed.
  3. Click to the option of "My Signature” and drag it.
  4. You will be given choice after selecting 'My Signature'. You can choose your drawn signature.
  5. Create your e-signature and drag 'Ok'.
  6. Select "Done".

You have successfully finish the PDF signing online . You can access your form and save it. Except for the e-sign choice CocoSign provides features, such as add field, invite to sign, combine documents, etc.

How to create an electronic signature for the Determination Descent Form in Chrome

Google Chrome is one of the most welcome browsers around the world, due to the accessibility of a large number of tools and extensions. Understanding the dire need of users, CocoSign is available as an extension to its users. It can be downloaded through the Google Chrome Web Store.

Follow these basic tips to generate an e-signature for your form in Google Chrome:

  1. Direct to the Web Store of Chrome and in the search CocoSign.
  2. In the search result, select the option of 'Add'.
  3. Now, sign in to your registered Google account.
  4. Click the link of the document and drag the option 'Open in e-sign'.
  5. Select the option of 'My Signature'.
  6. Create your signature and put it in the document where you favor.

After adding your e-sign, save your document or share with your team members. Furthermore, CocoSign provides its users the options to merge PDFs and add more than one signee.

How to create an electronic signature for the Determination Descent Form in Gmail?

Nowadays, businesses have altered their mode and evolved to being paperless. This involves the completing tasks through emails. You can easily e-sign the Determination Descent Form without logging out of your Gmail account.

Follow the tips below:

  1. Download the CocoSign extension from Google Chrome Web store.
  2. Open the document that needs to be e-signed.
  3. Select the "Sign” option and generate your signature.
  4. Select 'Done' and your signed document will be attached to your draft mail produced by the e-signature software of CocoSign.

The extension of CocoSign has solved problems for you. Try it today!

How to create an e-signature for the Determination Descent Form straight from your smartphone?

Smartphones have substantially replaced the PCs and laptops in the past 10 years. In order to solved problems for you, CocoSign helps finish your task via your personal phone.

A efficient internet connection is all you need on your phone and you can e-sign your Determination Descent Form using the tap of your finger. Follow the tips below:

  1. Direct to the website of CocoSign and create an account.
  2. Then, drag and upload the document that you need to get e-signed.
  3. Select the "My signature" option.
  4. Put down and apply your signature to the document.
  5. Take a look at the document and tap 'Done'.

It takes you a short time to add an e-signature to the Determination Descent Form from your phone. Get or share your form the way you want.

How to create an e-signature for the Determination Descent Form on iOS?

The iOS users would be pleased to know that CocoSign provides an iOS app to help out them. If an iOS user needs to e-sign the Determination Descent Form , utilize the CocoSign software with no doubt.

Here's guide add an electronic signature for the Determination Descent Form on iOS:

  1. Download the application from Apple Store.
  2. Register for an account either by your email address or via social account of Facebook or Google.
  3. Upload the document that needs to be signed.
  4. Click to the place where you want to sign and select the option 'Insert Signature'.
  5. Write your signature as you prefer and place it in the document.
  6. You can save it or upload the document on the Cloud.

How to create an electronic signature for the Determination Descent Form on Android?

The large popularity of Android phones users has given rise to the development of CocoSign for Android. You can download the software for your Android phone from Google Play Store.

You can add an e-signature for Determination Descent Form on Android following these tips:

  1. Login to the CocoSign account through email address, Facebook or Google account.
  2. Click your PDF file that needs to be signed electronically by selecting on the "+” icon.
  3. Direct to the place where you need to add your signature and generate it in a pop up window.
  4. Finalize and adjust it by selecting the '✓' symbol.
  5. Save the changes.
  6. Get and share your document, as desired.

Get CocoSign today to help out your business operation and save yourself a great amount of time and energy by signing your Determination Descent Form wherever.

Determination Descent Form FAQs

Some of the confused FAQs related to the Determination Descent Form are:

Need help? Contact support

Do military members have to pay any fee for leave or fiancee forms?

First off there are no fees for leaves or requests for leave in any branch of the United States military. Second there is no such thing as a fiancée form in the U.S. military. There is however a form for applying for a fiancée visa (K-1 Visa)that is available from the Immigration and Customs Service (Fiancé(e) Visas ) which would be processed by the U.S. State Department at a U.S. Consulate or Embassy overseas. However these fiancée visas are for foreigners wishing to enter the United States for the purpose of marriage and are valid for 90 days. They have nothing to do with the military and are Continue Reading

How can I fill out Google's intern host matching form to optimize my chances of receiving a match?

I was selected for a summer internship 2016. I tried to be very open while filling the preference form: I choose many products as my favorite products and I said I'm open about the team I want to join. I even was very open in the location and start date to get host matching interviews (I negotiated the start date in the interview until both me and my host were happy.) You could ask your recruiter to review your form (there are very cool and could help you a lot since they have a bigger experience). Do a search on the potential team. Before the interviews, try to find smart question that you are Continue Reading

How do I fill out the form of DU CIC? I couldn't find the link to fill out the form.

Just register on the admission portal and during registration you will get an option for the entrance based course. Just register there. There is no separate form for DU CIC.

How do you know if you need to fill out a 1099 form?

It can also be that he used the wrong form and will still be deducting taxes as he should be. Using the wrong form and doing the right thing isnt exactly a federal offense

How can I make it easier for users to fill out a form on mobile apps?

Make it fast. Ask them as few questions as possible (don't collect unnecessary information) and pre-populate as many fields as possible. Don't ask offputting questions where the respondent might have to enter sensitive personal information. If some users see you collecting sensitive information, they might not be ready to share that with you yet based on what you are offering, and they will think twice about completing the form.

When do I have to learn how to fill out a W-2 form?

While I did not study physics this is something that relates to my field as well. One thing to remember is the scope of the field which you are talking about. With physics it might seem narrower than History or Archaeology but I suspect that when you boil it down it isn’t. It would be impossible to cover everything in a subject even going all the way through to gaining a doctorate. The answer you got and posted up is very accurate and extremely good advice. What a lot of it boils down to in education (especially nowadays) is not so much teaching specific facts but teaching themes and how to find Continue Reading

Easier, Quicker, Safer eSignature Solution for SMBs and Professionals

No credit card required14 days free