Scanning optical homodyne detection of high-frequency picoscale resonances in cantilever and tuning fork sensors

Higher harmonic modes in nanoscale silicon cantilevers and microscale quartz tuning forks are detected and characterized using a custom scanning optical homodyne interferometer. Capable of both mass and force sensing, these resonators exhibit high-frequency harmonic motion content with picometer-scale amplitudes detected in a 2.5 MHz bandwidth, driven by ambient thermal radiation. Quartz tuning forks additionally display both in-plane and out-of-plane harmonics. The first six electronically detected resonances are matched to optically detected and mapped fork eigenmodes. Mass sensing experiments utilizing higher tuning fork modes indicate >6x sensitivity enhancement over fundamental mode operation.Comment: 3 pages, 3 figures, submitted to Applied Physics Letter

Filamentous Aggregates Are Fragmented by the Proteasome Holoenzyme.

Filamentous aggregates (fibrils) are regarded as the final stage in the assembly of amyloidogenic proteins and are formed in many neurodegenerative diseases. Accumulation of aggregates occurs as a result of an imbalance between their formation and removal. Here we use single-aggregate imaging to show that large fibrils assembled from full-length tau are substrates of the 26S proteasome holoenzyme, which fragments them into small aggregates. Interestingly, although degradation of monomeric tau is not inhibited by adenosine 5'-(3-thiotriphosphate) (ATPγS), fibril fragmentation is predominantly dependent on the ATPase activity of the proteasome. The proteasome holoenzyme also targets fibrils assembled from α-synuclein, suggesting that its fibril-fragmenting function may be a general mechanism. The fragmented species produced by the proteasome shows significant toxicity to human cell lines compared with intact fibrils. Together, our results indicate that the proteasome holoenzyme possesses a fragmentation function that disassembles large fibrils into smaller and more cytotoxic species.Wellcome Trust, Sir Henry Wellcome Fellowship (101585/Z/13/Z) to Yu Y

Direct Observation of Murine Prion Protein Replication in Vitro.

Prions are believed to propagate when an assembly of prion protein (PrP) enters a cell and replicates to produce two or more fibrils, leading to an exponential increase in PrP aggregate number with time. However, the molecular basis of this process has not yet been established in detail. Here, we use single-aggregate imaging to study fibril fragmentation and elongation of individual murine PrP aggregates from seeded aggregation in vitro. We found that PrP elongation occurs via a structural conversion from a PK-sensitive to PK-resistant conformer. Fibril fragmentation was found to be length-dependent and resulted in the formation of PK-sensitive fragments. Measurement of the rate constants for these processes also allowed us to predict a simple spreading model for aggregate propagation through the brain, assuming that doubling of the aggregate number is rate-limiting. In contrast, while α-synuclein aggregated by the same mechanism, it showed significantly slower elongation and fragmentation rate constants than PrP, leading to much slower replication rate. Overall, our study shows that fibril elongation with fragmentation are key molecular processes in PrP and α-synuclein aggregate replication, an important concept in prion biology, and also establishes a simple framework to start to determine the main factors that control the rate of prion and prion-like spreading in animals.J. C. S. is supported by a Cambridge Trust Scholarship and a Ministry of Education Technologies Incubation Scholarship, Republic of China (Taiwan). L. H. was supported by the Tsinghua University Initiative Scientific Research Program (Grants 20151080424) and the program of China Scholarships Council (CSC). A. M. T was supported in part by an MRC (NC3Rs) Project (Grant NC/K000462/1). G. M. and T. P. J. K. wish to acknowledge support from Sidney Sussex College Cambridge and the ERC grant PhysProt (337969). A. P. acknowledges funding from EPSRC (Grant EP/L027631/1). D. K. acknowledges funding from the Royal society and an ERC Advanced Grant (669237)

Optical Instrument Thermal Control on the Large Ultraviolet/Optical/Infrared Surveyor

The Large Ultraviolet/Optical/Infrared Surveyor (LUVOIR) is a multi-wavelength observatory commissioned by NASA as one of four large mission concept studies for the Astro2020 Decadal Survey. Two concepts are under study which bound a range of cost, risk, and scientific return: an 8-meter diameter unobscured segmented aperture primary mirror and a 15-meter segmented aperture primary mirror. Each concept carries with it an accompanying suite of instruments. The Extreme Coronagraph for Living Planetary Systems (ECLIPS) is a near-ultraviolet (NUV) / optical / near-infrared (NIR) coronagraph; the LUVOIR Ultraviolet Multi-object Spectrograph (LUMOS) provides multi-object imaging spectroscopy in the 100-400 nanometer ultraviolet (UV) range; and the High Definition Imager (HDI) is a wide field-of-view near-UV / optical / near-IR camera that can also perform astrometry. The 15-meter concept also contains an additional instrument, Pollux, which is a high-resolution UV spectro-polarimeter. While the observatory is nominally at a 270 Kelvin operational temperature, the requirements of imaging in both IR and UV require separate detectors operating at different temperature regimes, each with stringent thermal stability requirements. The change in observatory size requires two distinct thermal designs per instrument. In this current work, the thermal architecture is presented for each instrument suite. We describe here the efforts made to achieve the target operational temperatures and stabilities with passive thermal control methods. Additional discussion will focus on how these instrument thermal designs impact the overall system-level architecture of the observatory and indicate the thermal challenges for hardware implementation

L-Edge Spectroscopy of Dilute, Radiation-Sensitive Systems Using a Transition-Edge-Sensor Array

We present X-ray absorption spectroscopy and resonant inelastic X-ray scattering (RIXS) measurements on the iron L-edge of 0.5 mM aqueous ferricyanide. These measurements demonstrate the ability of high-throughput transition-edge-sensor (TES) spectrometers to access the rich soft X-ray (100-2000eV) spectroscopy regime for dilute and radiation-sensitive samples. Our low-concentration data are in agreement with high-concentration measurements recorded by conventional grating-based spectrometers. These results show that soft X-ray RIXS spectroscopy acquired by high-throughput TES spectrometers can be used to study the local electronic structure of dilute metal-centered complexes relevant to biology, chemistry and catalysis. In particular, TES spectrometers have a unique ability to characterize frozen solutions of radiation- and temperature-sensitive samples.Comment: 19 pages, 4 figure

WHOI Hawaii Ocean Timeseries Station (WHOTS) : WHOTS-9 2012 mooring turnaround cruise report

The Woods Hole Oceanographic Institution (WHOI) Hawaii Ocean Timeseries Site (WHOTS), 100 km north of Oahu, Hawaii, is intended to provide long-term, high-quality air-sea fluxes as a part of the NOAA Climate Observation Program. The WHOTS mooring also serves as a coordinated part of the Hawaii Ocean Timeseries (HOT) program, contributing to the goals of observing heat, fresh water and chemical fluxes at a site representative of the oligotrophic North Pacific Ocean. The approach is to maintain a surface mooring outfitted for meteorological and oceanographic measurements at a site near 22.75°N, 158°W by successive mooring turnarounds. These observations will be used to investigate air–sea interaction processes related to climate variability. This report documents recovery of the eighth WHOTS mooring (WHOTS-8) and deployment of the ninth mooring (WHOTS-9). Both moorings used Surlyn foam buoys as the surface element and were outfitted with two Air–Sea Interaction Meteorology (ASIMET) systems. Each ASIMET system measures, records, and transmits via Argos satellite the surface meteorological variables necessary to compute air–sea fluxes of heat, moisture and momentum. The upper 155 m of the moorings were outfitted with oceanographic sensors for the measurement of temperature, conductivity and velocity in a cooperative effort with R. Lukas of the University of Hawaii. A pCO2 system was installed on the buoys in cooperation with Chris Sabine at the Pacific Marine Environmental Laboratory. A set of radiometers were installed in cooperation with Sam Laney at WHOI. The WHOTS mooring turnaround was done on the NOAA ship Hi’ialakai by the Upper Ocean Processes Group of the Woods Hole Oceanographic Institution. The cruise took place between 12 and 19 June 2012. Operations began with deployment of the WHOTS-9 mooring on 13 June. This was followed by meteorological intercomparisons and CTDs. Recovery of the WHOTS-8 mooring took place on 16 June. This report describes these cruise operations, as well as some of the in-port operations and pre-cruise buoy preparations.Funding was provided by the National Oceanic and Atmospheric Administration under Grant No. NA09OAR4320129 and the Cooperative Institute for the North Atlantic Region (CINAR)

Soluble amyloid beta-containing aggregates are present throughout the brain at early stages of Alzheimer's disease.

Protein aggregation likely plays a key role in the initiation and spreading of Alzheimer's disease pathology through the brain. Soluble aggregates of amyloid beta are believed to play a key role in this process. However, the aggregates present in humans are still poorly characterized due to a lack of suitable methods required for characterizing the low concentration of heterogeneous aggregates present. We have used a variety of biophysical methods to characterize the aggregates present in human Alzheimer's disease brains at Braak stage III. We find soluble amyloid beta-containing aggregates in all regions of the brain up to 200 nm in length, capable of causing an inflammatory response. Rather than aggregates spreading through the brain as disease progresses, it appears that aggregation occurs all over the brain and that different brain regions are at earlier or later stages of the same process, with the later stages causing increased inflammation

Arachidonic acid mediates the formation of abundant alpha-helical multimers of alpha-synuclein

The protein alpha-synuclein (αS) self-assembles into toxic beta-sheet aggregates in Parkinson’s disease, while it is proposed that αS forms soluble alpha-helical multimers in healthy neurons. Here, we have made αS multimers in vitro using arachidonic acid (ARA), one of the most abundant fatty acids in the brain, and characterized them by a combination of bulk experiments and single-molecule Fӧrster resonance energy transfer (sm-FRET) measurements. The data suggest that ARA-induced oligomers are alpha-helical, resistant to fibril formation, more prone to disaggregation, enzymatic digestion and degradation by the 26S proteasome, and lead to lower neuronal damage and reduced activation of microglia compared to the oligomers formed in the absence of ARA. These multimers can be formed at physiologically-relevant concentrations, and pathological mutants of αS form less multimers than wild-type αS. Our work provides strong biophysical evidence for the formation of alpha-helical multimers of αS in the presence of a biologically relevant fatty acid, which may have a protective role with respect to the generation of beta-sheet toxic structures during αS fibrillation.M.I. acknowledges Dr. Tayyeb-Hussain Scholarship. L.T. has been recipient of a grant PAT Post Doc Outgoing 2009 7th Framework Program Marie Curie COFUND actions. C.D.H. and C.E.B. acknowledge funding from Alzheimer’s Research UK. Augustus Newman Foundation is acknowledged

Optical Instrument Thermal Control on the Large Ultraviolet/Optical/Infrared Surveyor

The Large Ultraviolet/Optical/Infrared Surveyor (LUVOIR) is a multi-wavelength observatory commissioned by NASA as one of four large mission concept studies for the Astro2020 Decadal Survey. Two concepts are under study which bound a range of cost, risk, and scientific return: an 8-meter diameter unobscured segmented aperture primary mirror and a 15-meter segmented aperture primary mirror. Each concept carries with it an accompanying suite of instruments. The Extreme Coronagraph for Living Planetary Systems (ECLIPS) is a near-ultraviolet (NUV) / optical / near-infrared (NIR) coronagraph; the LUVOIR Ultraviolet Multi-object Spectrograph (LUMOS) provides multi-object imaging spectroscopy in the 100-400 nanometer ultraviolet (UV) range; and the High Definition Imager (HDI) is a wide field-of-view near-UV / optical / near-IR camera that can also perform astrometry. The 15-meter concept also contains an additional instrument, Pollux, which is a high-resolution UV spectro-polarimeter. While the observatory is nominally at a 270 Kelvin operational temperature, the requirements of imaging in both IR and UV require separate detectors operating at different temperature regimes, each with stringent thermal stability requirements. The change in observatory size requires two distinct thermal designs per instrument. In this current work, the thermal architecture is presented for each instrument suite. We describe here the efforts made to achieve the target operational temperatures and stabilities with passive thermal control methods. Additional discussion will focus on how these instrument thermal designs impact the overall system-level architecture of the observatory and indicate the thermal challenges for hardware implementation