Low Gilbert damping and high perpendicular magnetic anisotropy in an Ir-coupled L10-FePd-based synthetic antiferromagnet.

Thin ferromagnetic films possessing perpendicular magnetic anisotropy derived from the crystal lattice can deliver the requisite magnetocrystalline anisotropy density for thermally stable magnetic memory and logic devices at the single-digit-nm lateral size. Here, we demonstrate that an epitaxial synthetic antiferromagnet can be formed from L10 FePd, a candidate material with large magnetocrystalline anisotropy energy, through insertion of an ultrathin Ir spacer. Tuning of the Ir spacer thickness leads to synthetic antiferromagnetically coupled FePd layers, with an interlayer exchange field upwards of 0.6 T combined with a perpendicular magnetic anisotropy energy of 0.95 MJ/m3 and a low Gilbert damping of 0.01. Temperature-dependent ferromagnetic resonance measurements show that the Gilbert damping is mostly insensitive to temperature over a range of 20 K up to 300 K. In FePd|Ir|FePd trilayers with lower interlayer exchange coupling, optic and acoustic dynamic ferromagnetic resonance modes are explored as a function of temperature. The ability to engineer low damping and large interlayer exchange coupling in FePd|Ir|FePd synthetic antiferromagnets with high perpendicular magnetic anisotropy could prove useful for high performance spintronic devices.

Single-cell profiling screen identifies microtubule-dependent reduction of variability in signaling.

Populations of isogenic cells often respond coherently to signals, despite differences in protein abundance and cell state. Previously, we uncovered processes in the Saccharomyces cerevisiae pheromone response system (PRS) that reduced cell-to-cell variability in signal strength and cellular response. Here, we screened 1,141 non-essential genes to identify 50 "variability genes". Most had distinct, separable effects on strength and variability of the PRS, defining these quantities as genetically distinct "axes" of system behavior. Three genes affected cytoplasmic microtubule function: BIM1, GIM2, and GIM4 We used genetic and chemical perturbations to show that, without microtubules, PRS output is reduced but variability is unaffected, while, when microtubules are present but their function is perturbed, output is sometimes lowered, but its variability is always high. The increased variability caused by microtubule perturbations required the PRS MAP kinase Fus3 and a process at or upstream of Ste5, the membrane-localized scaffold to which Fus3 must bind to be activated. Visualization of Ste5 localization dynamics demonstrated that perturbing microtubules destabilized Ste5 at the membrane signaling site. The fact that such microtubule perturbations cause aberrant fate and polarity decisions in mammals suggests that microtubule-dependent signal stabilization might also operate throughout metazoans.

Push-Pull and Feedback Mechanisms Can Align Signaling System Outputs with Inputs.

Many cell signaling systems, including the yeast pheromone response system, exhibit "dose-response alignment" (DoRA), in which output of one or more downstream steps closely matches the fraction of occupied receptors. DoRA can improve the fidelity of transmitted dose information. Here, we searched systematically for biochemical network topologies that produced DoRA. Most networks, including many containing feedback and feedforward loops, could not produce DoRA. However, networks including "push-pull" mechanisms, in which the active form of a signaling species stimulates downstream activity and the nominally inactive form reduces downstream activity, enabled perfect DoRA. Networks containing feedbacks enabled DoRA, but only if they also compared feedback to input and adjusted output to match. Our results establish push-pull as a non-feedback mechanism to align output with variable input and maximize information transfer in signaling systems. They also suggest genetic approaches to determine whether particular signaling systems use feedback or push-pull control.

Phasor plotting with frequency-domain flow cytometry.

Interest in time resolved flow cytometry is growing. In this paper, we collect time-resolved flow cytometry data and use it to create polar plots showing distributions that are a function of measured fluorescence decay rates from individual fluorescently-labeled cells and fluorescent microspheres. Phasor, or polar, graphics are commonly used in fluorescence lifetime imaging microscopy (FLIM). In FLIM measurements, the plotted points on a phasor graph represent the phase-shift and demodulation of the frequency-domain fluorescence signal collected by the imaging system for each image pixel. Here, we take a flow cytometry cell counting system, introduce into it frequency-domain optoelectronics, and process the data so that each point on a phasor plot represents the phase shift and demodulation of an individual cell or particle. In order to demonstrate the value of this technique, we show that phasor graphs can be used to discriminate among populations of (i) fluorescent microspheres, which are labeled with one fluorophore type; (ii) Chinese hamster ovary (CHO) cells labeled with one and two different fluorophore types; and (iii) Saccharomyces cerevisiae cells that express combinations of fluorescent proteins with different fluorescence lifetimes. The resulting phasor plots reveal differences in the fluorescence lifetimes within each sample and provide a distribution from which we can infer the number of cells expressing unique single or dual fluorescence lifetimes. These methods should facilitate analysis time resolved flow cytometry data to reveal complex fluorescence decay kinetics.

Measuring and sorting cell populations expressing isospectral fluorescent proteins with different fluorescence lifetimes.

Study of signal transduction in live cells benefits from the ability to visualize and quantify light emitted by fluorescent proteins (XFPs) fused to different signaling proteins. However, because cell signaling proteins are often present in small numbers, and because the XFPs themselves are poor fluorophores, the amount of emitted light, and the observable signal in these studies, is often small. An XFP's fluorescence lifetime contains additional information about the immediate environment of the fluorophore that can augment the information from its weak light signal. Here, we constructed and expressed in Saccharomyces cerevisiae variants of Teal Fluorescent Protein (TFP) and Citrine that were isospectral but had shorter fluorescence lifetimes, ∼ 1.5 ns vs ∼ 3 ns. We modified microscopic and flow cytometric instruments to measure fluorescence lifetimes in live cells. We developed digital hardware and a measure of lifetime called a "pseudophasor" that we could compute quickly enough to permit sorting by lifetime in flow. We used these abilities to sort mixtures of cells expressing TFP and the short-lifetime TFP variant into subpopulations that were respectively 97% and 94% pure. This work demonstrates the feasibility of using information about fluorescence lifetime to help quantify cell signaling in living cells at the high throughput provided by flow cytometry. Moreover, it demonstrates the feasibility of isolating and recovering subpopulations of cells with different XFP lifetimes for subsequent experimentation.

Confidence-accuracy relations for faces and scenes: roles of features and familiarity.

Using naturalistic scenes, we recently demonstrated that confidence-accuracy relations differ depending on whether recognition responses are based on memory for a specific feature or instead on general familiarity: When confidence is controlled for, accuracy is higher for familiarity-based than for feature-based responses. In the present experiment, we show that these results generalize to face recognition. Subjects studied photographs of scenes and faces presented for varying brief durations and received a recognition test on which they (1) indicated whether each picture was old or new, (2) rated their confidence in their response, and (3) indicated whether their response was based on memory for a feature or on general familiarity. For both stimulus types, subjects were more accurate and more confident for their feature-based than for their familiarity-based responses. However, when confidence was held constant, accuracy was higher for familiarity-based than for feature-based responses. These results demonstrate an important similarity between face and scene recognition and show that for both types of stimuli, confidence and accuracy are based on different information.

Hindsight bias from 3 to 95 years of age.

Upon learning the outcome to a problem, people tend to believe that they knew it all along (hindsight bias). Here, we report the first study to trace the development of hindsight bias across the life span. One hundred ninety-four participants aged 3 to 95 years completed 3 tasks designed to measure visual and verbal hindsight bias. All age groups demonstrated hindsight bias on all 3 tasks; however, preschoolers and older adults exhibited more bias than older children and younger adults. Multinomial processing tree analyses of these data revealed that preschoolers' enhanced hindsight bias resulted from them substituting the correct answer for their original answer in their recall (a qualitative error). Conversely, older adults' enhanced hindsight bias resulted from them forgetting their original answer and recalling an answer closer to, but not equal to, the correct answer (a quantitative error). We discuss these findings in relation to mechanisms of memory, perspective taking, theory of mind, and executive function.

Different confidence-accuracy relationships for feature-based and familiarity-based memories.

Participants studied naturalistic pictures presented for varying brief durations and then received a recognition test on which they indicated whether each picture was old or new and rated their confidence. In 1 experiment they indicated whether each "old"/"new" response was based on memory for a specific feature in the picture or instead on the picture's general familiarity; in another experiment, we defined pictures that tended to elicit feature versus familiarity responses. Thus, feature/familiarity was a dependent variable in 1 experiment and an independent variable in the other. In both experiments feature-based responses were more accurate than those that were familiarity based, and confidence and accuracy increased with duration for both response types. However, when confidence was controlled for, mean accuracy was higher for familiarity-based than for feature-based responses. The theoretical implication is that confidence and accuracy arise from different underlying information. The applied implication is that confidence differences should not be taken as implying accuracy differences when the phenomenal basis of the memory reports differ.