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All sourcesNASA NTRSDOE OSTI
NASA NTRS2004

Fluorescent Approaches to High Throughput Crystallography

X-ray crystallography remains the primary method for determining the structure of macromolecules. The first requirement is to have crystals, and obtaining them is often the rate-limiting step. The numbers of crystallization trials that are set up for any one protein for structural genomics, and the rate at which they are being set up, now overwhelm the ability for strictly human analysis of the results. Automated analysis methods are now being implemented with varying degrees of success, but these typically cannot reliably extract intermediate results. By covalently modifying a subpopulation, less than or = 1%, of a macromolecule solution with a fluorescent probe, the labeled material will add to a growing crystal as a microheterogeneous growth unit. Labeling procedures can be readily incorporated into the final stages of a macromolecules purification. The covalently attached probe will concentrate in the crystal relative to the solution, and under fluorescent illumination the crystals will show up as bright objects against a dark background. As crystalline packing is more dense than amorphous precipitate, the fluorescence intensity can be used as a guide in distinguishing different types of precipitated phases, even in the absence of obvious crystalline features, widening the available potential lead conditions in the absence of clear "bits." Non-protein structures, such as salt crystals, will not incorporate the probe and will not show up under fluorescent illumination. Also, brightly fluorescent crystals are readily found against less fluorescent precipitated phases, which under white light illumination may serve to obscure the crystals. Automated image analysis to find crystals should be greatly facilitated, without having to first define crystallization drop boundaries and by having the protein or protein structures all that show up. The trace fluorescently labeled crystals will also emit with sufficient intensity to aid in the automation of crystal alignment using relatively low cost optics, further increasing throughput at synchrotrons. This presentation will focus on the methodology for fluorescent labeling, the crystallization results, and the effects of the trace labeling on the crystal quality.

Minamitani, Elizabeth Forsythe↗
NASA NTRS2004

Fluorescent Approaches to High Throughput Crystallography

X-ray crystallography remains the primary method for determining the structure of macromolecules. The first requirement is to have crystals, and obtaining them is often the rate-limiting step. The numbers of crystallization trials that are set up for any one protein for structural genomics, and the rate at which they are being set up, now overwhelm the ability for strictly human analysis of the results. Automated analysis methods are now being implemented with varying degrees of success, but these typically can not reliably extract intermediate results. By covalently modifying a subpopulation, less than or = 1%, of a macromolecule solution with a fluorescent probe, the labeled material will add to a growing crystal as a microheterogeneous growth unit. Labeling procedures can be readily incorporated into the final stages of purification. The covalently attached probe will concentrate in the crystal relative to the solution, and under fluorescent illumination the crystals show up as bright objects against a dark background. As crystalline packing is more dense than amorphous precipitate, the fluorescence intensity can be used as a guide in distinguishing different types of precipitated phases, even in the absence of obvious crystalline features, widening the available potential lead conditions in the absence of clear "hits." Non-protein structures, such as salt crystals, will not incorporate the probe and will not show up under fluorescent illumination. Also, brightly fluorescent crystals are readily found against less fluorescent precipitated phases, which under white light illumination may serve to obscure the crystals. Automated image analysis to find crystals should be greatly facilitated, without having to first define crystallization drop boundaries and by having the protein or protein structures all that show up. The trace fluorescently labeled crystals will also emit with sufficient intensity to aid in the automation of crystal alignment using relatively low cost optics, further increasing throughput at synchrotrons. This presentation will focus on the methodology for fluorescent labeling, the crystallization results, and the effects of the trace labeling on the crystal quality.

Pusey, Marc L.↗
NASA NTRS2005

Fluorescent Approaches to High Throughput Crystallography

X-ray crystallography remains the primary method for determining the structure of macromolecules. The first requirement is to have crystals, and obtaining them is often the rate-limiting step. The numbers of crystallization trials that are set up for any one protein for structural genomics, and the rate at which they are being set up, now overwhelm the ability for strictly human analysis of the results. Automated analysis methods are now being implemented with varying degrees of success, but these typically cannot reliably extract intermediate results. By covalently modifying a subpopulation, 51%, of a macromolecule solution with a fluorescent probe, the labeled material will add to a growing crystal as a microheterogeneous growth unit. Labeling procedures can be readily incorporated into the final stages of purification. The covalently attached probe will concentrate in the crystal relative to the solution, and under fluorescent illumination the crystals show up as bright objects against a dark background. As crystalline packing is more dense than amorphous precipitate, the fluorescence intensity can be used as a guide in distinguishing different types of precipitated phases, even in the absence of obvious crystalline features, widening the available potential lead conditions in the absence of clear hits. Non-protein structures, such as salt crystals, will not incorporate the probe and will not show up under fluorescent illumination. Also, brightly fluorescent crystals are readily found against less fluorescent precipitated phases, which under white light illumination may serve to obscure the crystals. Automated image analysis to find crystals should be greatly facilitated, without having to first define crystallization drop boundaries and by having the protein or protein structures all that show up. The trace fluorescently labeled crystals will also emit with sufficient intensity to aid in the automation of crystal alignment using relatively low cost optics, further increasing throughput at synchrotrons. This presentation will focus on the methodology for fluorescent labeling, the crystallization results, and the effects of the trace labeling on the crystal quality.

Pusey, Marc L.↗
NASA NTRS2005

Fluorescent Approaches to High Throughput Crystallography

X-ray crystallography remains the primary method for determining the structure of macromolecules. The first requirement is to have crystals, and obtaining them is often the rate-limiting step. The numbers of crystallization trials that are set up for any one protein for structural genomics, and the rate at which they are being set up, now overwhelm the ability for strictly human analysis of the results. Automated analysis methods are now being implemented with varying degrees of success, but these typically cannot reliably extract intermediate results. By covalently modifying a subpopulation, less than or = 1 %, of a macromolecule solution with a fluorescent probe, the labeled material will add to a growing crystal as a microheterogeneous growth unit. Labeling procedures can be readily incorporated into the final stages of purification. The covalently attached probe will concentrate in the crystal relative to the solution, and under fluorescent illumination the crystals show up as bright objects against a dark background. As crystalline packing is more dense than amorphous precipitate, the fluorescence intensity can be used as a guide in distinguishing different types of precipitated phases, even in the absence of obvious crystalline features, widening the available potential lead conditions in the absence of clear "hits." Non-protein structures, such as salt crystals, will not incorporate the probe and will not show up under fluorescent illumination. Also, brightly fluorescent crystals are readily found against less fluorescent precipitated phases, which under white light illumination may serve to obscure the crystals. Automated image analysis to find crystals should be greatly facilitated, without having to first define crystallization drop boundaries and by having the protein or protein structures all that show up. The trace fluorescently labeled crystals will also emit with sufficient intensity to aid in the automation of crystal alignment using relatively low cost optics, further increasing throughput at synchrotrons. Preliminary experiments show that the presence of the fluorescent probe does not affect the nucleation process or the quality of the X-ray data obtained.

Pusey, Marc L.↗
NASA NTRS2006

Fluorescent Approaches to High Throughput Crystallography

We have shown that by covalently modifying a subpopulation, less than or equal to 1%, of a macromolecule with a fluorescent probe, the labeled material will add to a growing crystal as a microheterogeneous growth unit. Labeling procedures can be readily incorporated into the final stages of purification, and the presence of the probe at low concentrations does not affect the X-ray data quality or the crystallization behavior. The presence of the trace fluorescent label gives a number of advantages when used with high throughput crystallizations. The covalently attached probe will concentrate in the crystal relative to the solution, and under fluorescent illumination crystals show up as bright objects against a dark background. Non-protein structures, such as salt crystals, will not incorporate the probe and will not show up under fluorescent illumination. Brightly fluorescent crystals are readily found against less bright precipitated phases, which under white light illumination may obscure the crystals. Automated image analysis to find crystals should be greatly facilitated, without having to first define crystallization drop boundaries as the protein or protein structures is all that shows up. Fluorescence intensity is a faster search parameter, whether visually or by automated methods, than looking for crystalline features. We are now testing the use of high fluorescence intensity regions, in the absence of clear crystalline features or "hits", as a means for determining potential lead conditions. A working hypothesis is that kinetics leading to non-structured phases may overwhelm and trap more slowly formed ordered assemblies, which subsequently show up as regions of brighter fluorescence intensity. Preliminary experiments with test proteins have resulted in the extraction of a number of crystallization conditions from screening outcomes based solely on the presence of bright fluorescent regions. Subsequent experiments will test this approach using a wider range of proteins. The trace fluorescently labeled crystals will also emit with sufficient intensity to aid in the automation of crystal alignment using relatively low cost optics, further increasing throughput at synchrotrons.

Pusey, Marc L.↗
NASA NTRS2005

The flight instrument design for the Terrestrial Planet Finder Interferometer

This paper describes the opto-mechanical systems of the telescopes and beramcombiner spacecraft including the wavefront and optical path control devices and alignment systems. The mechanical layouts of the spacecraft are described along with the predicted performance of the structure in terms of thermal and vibration control. The layout of the control system is also described.

interferometry↗
NASA NTRS2006

Fluorescent Applications to Crystallization

By covalently modifying a subpopulation, less than or equal to 1%, of a macromolecule with a fluorescent probe, the labeled material will add to a growing crystal as a microheterogeneous growth unit. Labeling procedures can be readily incorporated into the final stages of purification, and tests with model proteins have shown that labeling u to 5 percent of the protein molecules does not affect the X-ray data quality obtained . The presence of the trace fluorescent label gives a number of advantages. Since the label is covalently attached to the protein molecules, it "tracks" the protein s response to the crystallization conditions. The covalently attached probe will concentrate in the crystal relative to the solution, and under fluorescent illumination crystals show up as bright objects against a darker background. Non-protein structures, such as salt crystals, do not show up under fluorescent illumination. Crystals have the highest protein concentration and are readily observed against less bright precipitated phases, which under white light illumination may obscure the crystals. Automated image analysis to find crystals should be greatly facilitated, without having to first define crystallization drop boundaries as the protein or protein structures is all that shows up. Fluorescence intensity is a faster search parameter, whether visually or by automated methods, than looking for crystalline features. Preliminary tests, using model proteins, indicates that we can use high fluorescence intensity regions, in the absence of clear crystalline features or "hits", as a means for determining potential lead conditions. A working hypothesis is that more rapid amorphous precipitation kinetics may overwhelm and trap more slowly formed ordered assemblies, which subsequently show up as regions of brighter fluorescence intensity. Experiments are now being carried out to test this approach using a wider range, of proteins. The trace fluorescently labeled crystals will also emit with sufficient intensity to aid in the automation of crystal alignment using relatively low cost optics, further increasing throughput at synchrotrons.

Pusey, Marc L.↗
NASA NTRS2010

NASA Tech Briefs, September 2010

Topics covered include: Instrument for Measuring Thermal Conductivity of Materials at Low Temperatures; Multi-Axis Accelerometer Calibration System; Pupil Alignment Measuring Technique and Alignment Reference for Instruments or Optical Systems; Autonomous System for Monitoring the Integrity of Composite Fan Housings; A Safe, Self-Calibrating, Wireless System for Measuring Volume of Any Fuel at Non-Horizontal Orientation; Adaptation of the Camera Link Interface for Flight-Instrument Applications; High-Performance CCSDS Encapsulation Service Implementation in FPGA; High-Performance CCSDS AOS Protocol Implementation in FPGA; Advanced Flip Chips in Extreme Temperature Environments; Diffuse-Illumination Systems for Growing Plants; Microwave Plasma Hydrogen Recovery System; Producing Hydrogen by Plasma Pyrolysis of Methane; Self-Deployable Membrane Structures; Reactivation of a Tin-Oxide-Containing Catalys; Functionalization of Single-Wall Carbon Nanotubes by Photo-Oxidation; Miniature Piezoelectric Macro-Mass Balance; Acoustic Liner for Turbomachinery Applications; Metering Gas Strut for Separating Rocket Stages; Large-Flow-Area Flow-Selective Liquid/Gas Separator; Counterflowing Jet Subsystem Design; Water Tank with Capillary Air/Liquid Separation; True Shear Parallel Plate Viscometer; Focusing Diffraction Grating Element with Aberration Control; Universal Millimeter-Wave Radar Front End; Mode Selection for a Single-Frequency Fiber Laser; Qualification and Selection of Flight Diode Lasers for Space Applications; Plenoptic Imager for Automated Surface Navigation; Maglev Facility for Simulating Variable Gravity; Hybrid AlGaN-SiC Avalanche Photodiode for Deep-UV Photon Detection; High-Speed Operation of Interband Cascade Lasers; 3D GeoWall Analysis System for Shuttle External Tank Foreign Object Debris Events; Charge-Spot Model for Electrostatic Forces in Simulation of Fine Particulates; Hidden Statistics Approach to Quantum Simulations; Reconstituted Three-Dimensional Interactive Imaging; Determining Atmospheric-Density Profile of Titan; Digital Microfluidics Sample Analyzer; Radiation Protection Using Carbon Nanotube Derivatives; Process to Selectively Distinguish Viable from Non-Viable Bacterial Cells; and TEAMS Model Analyzer.

Source record↗
NASA NTRS2019

Analysis of the Global Microwave Polarization Data of Clouds

Two NASA microwave radiometers, the satellite-borne GPM (Global Precipitation Measurement) Microwave Imager (GMI) and the aircraft-borne CoSMIR (the Conical Scanning Millimeter-wave Imaging Radiometer), measure vertically- and horizontally-polarized microwaves emitted by cloud particles and the Earth below, providing unique information on ice crystal properties in clouds. Their data reveal that non-spherical ice crystals are common and they fall in a preferred horizontally aligned orientation in convective and optically thick clouds especially near cloud top. A bin (particle-size-resolving) microphysical model with an ice crystal shape representation is developed to simulate the evolution of ice crystal properties (i.e., size, shape and orientation), where the radiation effect on microphysics (REM) is taken into account. Since REM represents the effect of all (e.g., both infrared and solar) radiation on ice crystal temperature, it relies upon the ice crystal proprieties that determine how an ice crystal receives radiation. Definitely, REM is different from the radiative effects that cause sensitivity at the microwave frequencies in the GPM and CoSMIR observations. Model results show that horizontally-oriented ice crystals grow faster than vertically-oriented (or spherical) ones due to REM. When both horizontally- and vertically-oriented ice crystals coexist in an air parcel, the model results show that the former grow by vapor deposition whereas the latter shrink by sublimation and disappear eventually. These modeling results are supported by the GMI data and the CoSMIR observations from MC3E (Midlatitude Continental Convective Clouds Experiment) and OLYMPEX (Olympic Mountains Experiment) on the prevalence of horizontally-oriented ice crystals. Moreover, the REM-induced precipitation explains the CloudSat observations of rare thin clouds in the tropical mid-troposphere as well as the common diamond dust in the high latitudes.

Xiping Zeng↗
NASA NTRS2017

Discovery Deep Space Optical Communications (DSOC) transceiver

NASA’s 22 cm diameter Deep Space Optical Communications (DSOC) Transceiver is designed to provide a bi-directional optical link between a spacecraft in the inner solar system and an Earth-based optical ground station. This design, optimized for operation across a wide range of illumination conditions, is focused on minimizing blinding from stray light, and providing reliable, accurate attitude information to point its narrow communication beam accurately to the future location of the ground terminal. Though our transceiver will transmit in the 1550 nm waveband and receive in the 1064 nm waveband, the system design relies heavily on reflective optical elements, extending flexibility to be modified for use at different wavebands. The design makes use of common path propagation among transmit, receive and pointing verification optical channels to maintain precise alignment among its components, and to naturally correct for element misalignment resulting from launch or thermal element perturbations. This paper presents the results of trade studies showing the evolution of the design, unique operational characteristics of the design, elements that help to maintain minimal stray light contamination, and preliminary results from development and initial testing of a functional aluminum test model.

Roberts, W. Thomas↗
DOE OSTI2024

Six degrees of freedom compliant flexure and autonomous assembly using same

A flexure having six degrees of freedom is disclosed. The constraint and compliance of the flexure in all six degrees of freedom may be independently tuned for an application. The flexure may include various combinations of sections including, for example, straight arm and curvilinear sections. The constraint and compliance of the flexure is determined by the geometry, cross-sectional area, cross-sectional shape, and material used to form the various sections. A coupling mechanism and a corresponding method for using the coupling mechanism for automated or autonomous assembly of elements using tuned flexures is disclosed. The automated or autonomous assembly of elements employs a coupling mechanism having a motor driven grip and a coupler, in which the coupler includes a six degrees of freedom flexure. The motor driven grip and coupler may optionally include electrical or optical interconnections and self-aligning features.

Gillund, Daniel Paul↗
NASA NTRS1991

Design of a high-gain laser diode-array pumped Nd:YAG Alternating Precessive Slab Amplifier (APS-Amplifier)

In the design of space qualifiable laser systems for ranging and altimetry, such as NASA's Geodynamic Laser Ranging System (GLRS), the transmitter must be kept small, powerful yet efficient, and must consist of as few components as possible. A novel preamplifier design is examined which requires no external beam steering optics, yielding a compact component with simple alignment procedures. The gains achieved are comparable to multipass zigzag amplifiers using two or more sets of external optics for extra passes through the amplifying medium.

Coyle, D. Barry↗
NASA NTRS1991

Design of a high-gain laser diode-array pumped Nd:YAG alternating precessive slab amplifier (APS amplifier)

In the design of space-qualifiable laser systems for ranging and altimetry, such as NASA's Geodynamic Laser Ranging System (GLRS), the transmitter must be kept small, powerful yet efficient, and must consist of as few components as possible. A novel preamplifier design is examined which requires no external beam steering optics, yielding a compact component with simple alignment procedures. The gains achieved are comparable to multipass zigzag amplifiers using two or more sets of external optics for extra passes through the amplifying medium.

Coyle, D. B.↗
NASA NTRS2012

Telescope Alignment From Sparsely Sampled Wavefront Measurements Over Pupil Subapertures

Alignment of two-element telescopes is a classic problem. During recent integration and test of the Space Interferometry Mission s (SIM s) Astrometric Beam Combiner (ABC), the innovators were faced with aligning two such telescope subsystems in the presence of a further complication: only two small subapertures in each telescope s pupil were accessible for measuring the wavefront with a Fizeau interferometer. This meant that the familiar aberrations that might be interpreted to infer system misalignments could be viewed only over small sub-regions of the pupil, making them hard to recognize. Further, there was no contiguous surface of the pupil connecting these two subapertures, so relative phase piston information was lost; the underlying full-aperture aberrations therefore had an additional degree of ambiguity. The solution presented here is to recognize that, in the absence of phase piston, the Zygo measurements primarily provide phase tilt in the subaperture windows of interest. Because these windows are small and situated far from the center of the (inaccessible) unobscured full aperture, any aberrations that are higher-order than tilt will be extremely high-order on the full aperture, and so not necessary or helpful to the alignment. Knowledge of the telescope s optical prescription allows straightforward evaluation of sensitivities (subap mode strength per unit full-aperture aberration), and these can be used in a predictive matrix approach to move with assurance to an aligned state. The technique is novel in every operational way compared to the standard approach of alignment based on full-aperture aberrations or searching for best rms wavefront. This approach is closely grounded in the observable quantities most appropriate to the problem. It is also more intuitive than inverting full phase maps (or subaperture Zernike spectra) with a ray-tracing program, which must certainly work in principle, but in practice met with limited success. Even if such classical alignment techniques became practical, the techniques reported here form a reassuringly transparent and intuitive check on the course of the alignment with very little computational effort.

Bloemhof, Eric E.↗
NASA NTRS2004

A Wafer-Bonded, Floating Element Shear-Stress Sensor Using a Geometric Moire Optical Transduction Technique

This paper presents a geometric Moir optical-based floating-element shear stress sensor for wind tunnel turbulence measurements. The sensor was fabricated using an aligned wafer-bond/thin-back process producing optical gratings on the backside of a floating element and on the top surface of the support wafer. Measured results indicate a static sensitivity of 0.26 microns/Pa, a resonant frequency of 1.7 kHz, and a noise floor of 6.2 mPa/(square root)Hz.

Horowitz, Stephen↗
NASA NTRS1999

SPARCLE Optical System Design and Operational Characteristics

The SPAce Readiness Coherent Lidar Experiment (SPARCLE) is the first demonstration of a coherent Doppler wind lidar in space. SPARCLE will be flown aboard a space shuttle In the middle part of 2001 as a stepping stone towards the development and deployment of a long-life-time operational instrument in the later part of next decade. SPARCLE is an ambitious project that is intended to evaluate the suitability of coherent lidar for wind measurements, demonstrate the maturity of the technology for space application, and provide a useable data set for model development and validation. This paper describes the SPARCLE's optical system design, fabrication methods, assembly and alignment techniques, and its anticipated operational characteristics. Coherent detection is highly sensitive to aberrations in the signal phase front, and to relative alignment between the signal and the local oscillator beams. Consequently, the performance of coherent lidars is usually limited by the optical quality of the transmitter/receiver optical system. For SPARCLE having a relatively large aperture (25 cm) and a very long operating range (400 km), compared to the previously developed 2-micron coherent lidars, the optical performance requirements are even more stringent. In addition with stringent performance requirements, the physical and environment constraints associated with this instrument further challenge the limit of optical fabrication technologies.

Amzajerdian, Farzin↗
NASA NTRS1973

Investigation of the dielectric waveguide modes in homostructure GaAs laser.

The investigation of the propagation modes in a homostructure GaAs laser and particularly of the spatial displacement of the TE and TM mode is reported. For this investigation, a homostructure GaAs laser was operated in an external optical cavity and experiments were performed where the resonator alignment was varied to displace the image of the p-n junction, formed in the optical cavity, from the p-n junction of the laser diode. The peak power of the laser radiation (which is in the TEM (sub 00) mode), the delay, the optical waveform, and the polarization were measured at room temperature for different cavity alignments. For the evaluation of the propagation modes the experimental results are correlated with the computed field distributions of the modes in a three-layer dielectric waveguide.

Philipp-Rutz, E. M.↗
NASA NTRS2006

Phase Sensor for Aligning a Segmented Telescope Mirror

A phase sensor has been developed for use in aligning a segmented telescope mirror to within a fraction of a wavelength in piston. (As used here, piston signifies displacement of a mirror segment along the optical axis of the telescope.) Such precise alignment is necessary in order to realize the full benefit of the large aperture achievable through segmentation. This phase sensor is achromatic. It is based on two-wavelength shearing interferometry, and can be modified to utilize an extended or broad-band (e.g., white) light source. The sensor optics include a ruled diffraction grating and an imaging lens. The sensor can measure the piston shift between segments as well as aberrations of the segments. It can measure the surface error of an individual segment, making it possible to compensate for the error with optimal amount(s) of piston and/or tilt. The precise capture range of the sensor depends partly on the telescope design; the largest relative piston shifts measurable by use of this sensor are of the order of 100 m. The accuracy of the sensor also depends partly on the telescope design; in general, the accuracy is sufficient to enable alignment to within approximately half a wavelength. The interferometric image is digitized and processed by a simple algorithm in real time, and the output of the algorithm can be used to maintain alignment in real time, even in the presence of atmospheric turbulence. The sensor is robust. Through calibration, it can be made insensitive to (and, hence, tolerant of) misalignments and aberrations of its own optics, most aberrations of the telescope as a whole (in contradistinction to aberrations of individual segments), and most aberrations introduced by atmospheric turbulence

Stahl, Philip↗