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NASA NTRS1995

Sizing of colloidal particle and protein molecules in a hanging fluid drop

We report non-invasive particle size measurements of polystyrene latex colloidal particles and bovine serum albumin (BSA) protein molecules suspended in tiny hanging fluid drops of 30 micro-Liter volume using a newly designed fiber optic probe. The probe is based upon the principles of the technique of dynamic light scattering (DLS). The motivation for this work comes from growing protein crystals in outer space. Protein crystals have been grown previously in hanging drops in microgravity experiments on-board the space shuttle orbiter. However, obtaining quantitative information on nucleation and growth of the protein crystals in real time has always been a desired goal, but hitherto not achieved. Several protein researchers have shown interest in using DLS to monitor crystal growth process in a droplet, but elaborate instrumentation and optical alignment problems have made in-situ applications difficult. We demonstrate that such an experiment is now possible. Our system offers fast (5 seconds) determination of particle size, utilize safe levels of very low laser power (less than or equal to 0.2 mW), a small scattering volume (approximately 2 x 10(exp -5) cu mm) and high spatial coherence (Beta) values. This is a major step forward when compared to currently available DLS systems.

Ansari, Rafat R.↗

NASA NTRS1996

Chain Dynamics in a Dilute Magnetorheological Fluid

The structure, formation, and dynamics of dilute, mono-dispersive ferrofluid emulsions in an external magnetic field have been investigated using dynamic light scattering techniques. In the absence of the magnetic field, the emulsion particles are randomly distributed and behave like hard spheres in Brownian motion. An applied magnetic field induces a magnetic dipole moment in each particle. Dipolar interactions between particles align them into chains where correlation functions show two decay processes. The short-time decay shows the motion of straight chains as a whole where the apparent chain length increases with the applied magnetic field and the particle volume fraction. Good scaling results are obtained showing that the apparent chain length grows with time following a power law with exponent of 0.6 and depends on the applied field, particle volume fraction, and diffusion constant of the particles. The long-time decay in the correlation function shows oscillation when the chains reach a certain length with time and stiffness with threshold field This result shows that chains not only fluctuate, but move in a periodic motion with a frequency of 364 Hz at lambda = 15. It may suggest the existence of phonons. This work is the first step in the understanding of the structure formation, especially chain coarsening mechanism, of magnetorheological (MR) fluids at higher volume fractions.

Liu, Jing↗

NASA NTRS2000

Over-Expression, Purification and Crystallization of Human Dihydrolipoamide Dehydrogenase

Dehydrolipoamide dehydrogenase (E3; dihydrolipoan-tide:NAD+ oxidoreductase, EC 1.8.1.4) is a common catalytic component found in pyruvate dehydrogenase complex, alpha-ketoglutarate dehydrogenase complex, and branched-chain cc-keto acid dehydrogenase complex. E3 is also a component (referred to as L protein) of the glycine cleavage system in bacterial metabolism (2). Active E3 forms a homodimer with four distinctive subdomain structures (FAD binding, NAD+ binding, central and interface domains) with non-covalently but tightly bound FAD in the holoenzyme. Deduced amino acids from cloned full-length human E3 gene showed a total of 509 amino acids with a leader sequence (N-terminal 35 amino acids) that is excised (mature form) during transportation of expressed E3 into mitochondria membrane. So far, three-dimensional structure of human E3 has not been reported. Our effort to achieve the elucidation of the X-ray crystal structure of human E3 will be presented. Recombinant pPROEX-1 expression vector (from GIBCO BRL Life Technologies) having the human E3 gene without leader sequence was constructed by Polymerase Chain Reaction (PCR) and subsequent ligation, and cloned in E.coli XL1-Blue by transformation. Since pPROEX-1 vector has an internal His-tag (six histidine peptide) located at the upstream region of a multicloning site, one-step affinity purification of E3 using nickelnitriloacetic acid (Ni-NTA) agarose resin, which has a strong affinity to His-tag, was feasible. Also a seven-amino-acid spacer peptide and a recombinant tobacco etch virus protease recognition site (seven amino acids peptide) found between His-tag and first amino acid of expressed E3 facilitated the cleavage of His-tag from E3 after the affinity purification. By IPTG induction, ca. 15 mg of human E3 (mature form) was obtained from 1L LB culture with overnight incubation at 25C. Over 98% of purity of E3 from one-step Ni-NTA agarose affinity purification was confirmed by SDS-PAGE analysis. For crystallization, E3 samples were prepared with and without His-tag. To minimize the aggregation of E3, apo- and holo- forms of E3s were tested, as well as a mutated E3. Dynamic light scattering measurements revealed that the E3 preparations without His-tag and substrate are highly monodispersive with regard to homodimers. Consequent crystallization trials of this E3 preparation led to single crystals of E3 grown by the vapor diffusion method. Crystals were obtained within a few days from solution containing poly (ethylene glycol) monomethyl ether 5000 as a precipitant. Autoindexing and integration of the X-ray diffraction data showed that E3 crystals belong to an orthorhombic system with unit cell parameters a-- 123. 1, b= 165.3 and c=214.3A. Further optimization of protein preparation and crystallization experiments for the structural determination will be discussed.

Hong, Y. S.↗

NASA NTRS2000

Monitoring Astronaut Health at the Nanoscale Cellular Level Through the Eye

A user friendly goggles-like head-mounted device equipped with a suite of instruments for several non-invasive and quantitative medical evaluation of the eye, skin, and brain is desired for monitoring the health of astronauts during space travel and exploration of neighboring and distant planets. Real-time non-invasive evaluation of the different structures within the above organs can provide indices of the health of not just these organs, but the entire body. The techniques such as dynamic light scattering (for the early detection of uveitis, cholesterol levels, cataract, changes in the vitreous and possibly Alzheimer's disease), corneal autofluorescence (to assess extracellular matrix biology e.g., in diabetes), optical activity measurements (of anterior ocular fluid to evaluate blood-glucose levels), laser Doppler velocimetry (to assess retinal, optic nerve, and choroidal blood flow), reflectometry/oximetry (for assessing ocular and central nervous system oxygen metabolism), optical coherence tomography (to determine retinal tissue microstructure) and possibly scanning laser technology (for intraocular tissue imaging and scanning) will he integrated into this compact device. Skin sensors will also be mounted on the portion of the device in contact with the periocular region. This will enable monitoring of body temperature, EEG, and electrolyte status. This device will monitor astronaut health during long-duration space travel by detecting aberrations from pre-established "nonns", enabling prompt diagnosis and possibly the initiation of early preventative/curative therapy. The non-invasive nature of the device technologies permits frequent repetition of tests, enabling real-time complete crew health monitoring. This device may ultimately be useful in tele-medicine to bring modern healthcare to under-served areas on Earth as well as in so-called "advanced" care settings (e.g. diabetes in the USA).

Ansari, Rafat R.↗

NASA NTRS2001

Testing Microgravity Flight Hardware Concepts on the NASA KC-135

This paper provides an overview of utilizing the NASA KC-135 Reduced Gravity Aircraft for the Foam Optics and Mechanics (FOAM) microgravity flight project. The FOAM science requirements are summarized, and the KC-135 test-rig used to test hardware concepts designed to meet the requirements are described. Preliminary results regarding foam dispensing, foam/surface slip tests, and dynamic light scattering data are discussed in support of the flight hardware development for the FOAM experiment.

Motil, Susan M.↗

NASA NTRS2002

Non-Invasive, Quantitative, and Remote Detection of Early Radiation Cataracts for Applications in Bio-Astronautics and Bio-Informatics

Human exploration of Mars may be a possibility in the next twenty years. Maintaining good vision is an essential aspect of achieving a successful mission. Continuous radiation exposure is a risk factor for radiation-induced cataracts in astronauts. A compact device based on the technique of dynamic light scattering (DLS) is designed for monitoring an astronaut's ocular health during long-duration space travel. Preliminary data on the simulated effects of ionizing radiation exposure to the ocular tissues of nonhuman animals and results on the sensitivity of DLS over established clinical procedures in investigating cataracts are presented. This capability of early diagnosis, unmatched by any other clinical technique in use today, may enable prompt initiation of preventive/curative therapy. An internet web based system integrating photon correlation data and controlling the hardware to monitor cataract development in vivo at a remote site in real time (teleophthalmology) is currently being developed. Cataract studies on-board the International Space Station (ISS) will be helpful in designing better protective radiation shields for future space vehicles and space suits.

Ansari, Rafat R.↗

NASA NTRS2002

Non-Invasive Health Diagnostics using Eye as a 'Window to the Body'

As a 'window to the body', the eye offers the opportunity to use light in various forms to detect ocular and systemic abnormalities long before clinical symptoms appear and help develop preventative/therapeutic countermeasures early. The effects of space travel on human body are similar to those of normal aging. For example, radiation exposure in space could lead to formation of cataracts and cancer by damaging the DNA and causing gene mutation. Additionally, the zero-gravity environment causes fluid shifts in the upper extremities of the body and changes the way blood flows and organ system performs. Here on Earth, cataract, age-related macular degeneration (AMD), diabetic retinopathy (DR), and glaucoma are major eye diseases and are expected to double in next two decades. To detect, prevent, and treat untoward effects of prolonged space travel in real-time requires the development of non-invasive diagnostic technologies that are compact and powerful. We are developing fiber-optic sensors to evaluate the ocular tissues in health, aging, and disease employing the techniques of dynamic light scattering (cataract, uveitis, Alzheimer's, glaucoma, DR, radiation damage, refractive surgery outcomes), auto-fluorescence (aging, DR), laser-Doppler flowmetry (choroidal blood flow), Raman spectroscopy (AMD), polarimetry (diabetes), and retinal oximetry (occult blood loss). The non-invasive feature of these technologies integrated in a head-mounted/goggles-like device permits frequent repetition of tests, enabling evaluation of the results to therapy that may ultimately be useful in various telemedicine applications on Earth and in space.

Ansari, Rafat R.↗

NASA NTRS2004

Dissemination of information about the technologies of the Vision Research Lab through the World Wide Web

The Vision Research Lab at NASA John Glenn Research Center is headed by Dr. Rafat Ansari. Dr. Ansari and other researchers have developed technologies that primarily use laser and fiber optics to non-invasively detect different ailments and diseases of the eye. One of my goals as a LERCIP intern and ACCESS scholar for the 2004 summer is to inform other NASA employees, researchers and the general public about these technologies through the development of a website. The website incorporates the theme that the eye is a window to the body. Thus by investigating the processes of the eye, we can better understand and diagnosis different ailments and diseases. These ailments occur in not only earth bound humans, but astronauts as well as a result of exposure to elevated levels of radiation and microgravity conditions. Thus the technologies being developed at the Vision Research Lab are invaluable to humans on Earth in addition to those astronauts in space. One of my first goals was to research the technologies being developed at the lab. The first several days were spent immersing myself in the various articles, journals and reports about the theories behind Dynamic Light Scattering, Laser Doppler Flowmetry, Autofluoresence, Raman Spectroscopy, Polarimetry and Oximetry. Interviews with the other researchers proved invaluable to help understand these theories as well gain hands on experience with the devices being developed using these technologies. The rest of the Vision Research Team and I sat down and discussed how the overall website should be presented. Combining this information with the knowledge of the theories and applications of the hardware being developed, I worked out different ideas to present this information. I quickly learned Paint Shop Pro 8 and FrontPage 2002, as well as using online tutorials and other resources to help design an effective website. The Vision Research Lab website incorporates the anatomy and physiology of the eye, different diseases that affect the eye and the technologies being develop at the lab to help diagnosis these diseases. It also includes background information on Dr. Ansari as well as other researchers involved in the lab and it includes segments on patents, awards and achievements. There are links to help viewers navigate to internal and external websites to further investigate different ideas and hrther understand the implications of these technologies at being developed.

Dorais, Christopher M.↗

NASA NTRS2000

Applications in Bioastronautics and Bioinformatics: Early Radiation Cataracts Detected by Noninvasive, Quantitative, and Remote Means

Human exploration of Mars is a key goal in NASA's exploration planning in the next 20 years. Maintaining crew health and good vision is certainly an important aspect of achieving a successful mission. Continuous radiation exposure is a risk factor for radiation-induced cataracts in astronauts because radiation exposure in space travel has the potential of accelerating the aging process (ref. 1). A patented compact device (ref. 2) based on the technique of dynamic light scattering (DLS) was designed for monitoring an astronaut's ocular health during long-duration space travel. This capability of early diagnosis, unmatched by any other clinical technique in use today, may enable prompt initiation of preventive/curative therapy. An Internet web-based system integrating photon correlation data and controlling the hardware to monitor cataract development in vivo at a remote site in real time (teleophthalmology) is currently being developed. The new technology detects cataracts very early (at the molecular level). Cataract studies onboard the International Space Station will be helpful in quantifying any adverse effect of radiation to ocular health. The normal lens in a human eye, situated behind the cornea, is a transparent tissue. It contains 35 wt % protein and 65 wt % water. Aging, disease (e.g., diabetes), smoking, dehydration, malnutrition, and exposure to ultraviolet light and ionizing radiation can cause agglomeration of the lens proteins. Protein aggregation can take place anywhere in the lens, causing lens opacity. The aggregation and opacification could produce nuclear (central portion of the lens) or cortical (peripheral) cataracts. Nuclear and posterior subcapsular (the membrane's capsule surrounds the whole lens) cataracts, being on the visual optical axis of the eye, cause visual impairment that can finally lead to blindness. The lens proteins, in their native state, are small in size. As a cataract develops, this size grows from a few nanometers (transparent) to several micrometers (cloudy). Ansari and Datiles have shown that DLS can detect cataracts at least two to three orders of magnitude earlier noninvasively and quantitatively than the best imaging (Scheimpflug) techniques in clinical use today (ref. 3).

Ansari, Rafat R.↗

NASA NTRS2006

Ultrasonication of Bismuth Telluride Nanocrystals Fabricated by Solvothermal Method

The objective of this study is to evaluate the effect of ultrasonication on bismuth telluride nanocrystals prepared by solvothermal method. In this study, a low dimensional nanocrystal of bismuth telluride (Bi2Te3) was synthesized by a solvothermal process in an autoclave at 180 C and 200 psi. During the solvothermal reaction, organic surfactants effectively prevented unwanted aggregation of nanocrystals in a selected solvent while controlling the shape of the nanocrystal. The atomic ratio of bismuth and tellurium was determined by energy dispersive spectroscopy (EDS). The cavitational energy created by the ultrasonic probe was varied by the ultrasonication process time, while power amplitude remained constant. The nanocrystal size and its size distribution were measured by field emission scanning electron microscopy (FESEM) and a dynamic light scattering system. When the ultrasonication time increased, the average size of bismuth telluride nanocrystal gradually increased due to the direct collision of nanocrystals. The polydispersity of the nanocrystals showed a minimum when the ultrasonication was applied for 5 min. Keywords: bismuth telluride, nanocrystal, low-dimensional, ultrasonication, solvothermal

Chu, Sang-Hyon↗

NASA NTRS2021

Water/Oil Emulsions with Controlled Droplet Sizes for In Vitro Selection Experiments

In the early history of life, RNA might have had many catalytic functions as ribozymes that do not exist today. To explore this possibility, catalytically active RNAs can be identified by in vitro selection experiments. Some of these experiments are best performed in nanodroplets to prevent diffusion between individual RNA sequences. In order to explore the suitability for the large-scale in emulsio selection of water-in-oil emulsions made by passing a mixture of mineral oil, the emulsifier ABIL-EM90, and a few percent of an aqueous phase through a microfluidizer, we used dynamic light scattering to characterize the size of aqueous droplets dispersed throughout the oil. We found that seven or more passes through the microfluidizer at 8000 psi with close to half molar inorganic salts and 10% polyethylene glycol produced droplets with sizes below 100 nm that were ideal for our purposes. We also identified conditions that would produce larger or smaller droplets, and we demonstrate that the emulsions are stable over weeks and months, which is desirable for different types of in vitro selection experiments.

Douglas Magde↗

NASA NTRS1993

Measuring the dynamics of structural changes in biological macromolecules from light scattering data

Examining techniques to study the dynamics of structural changes in various molecules has been an ongoing goal of the space program. Knowing how these phenomena occur in biological systems is fundamental to understanding what is necessary for life to remain functional in the space environment. A hierarchy of biological organization is functionally described when cells join together small organic molecules to form larger and more complex molecules. Characterizing the architecture of a particular macromolecule helps determine how that molecule works in the living cell and is basic to the diversity of life. Understanding this arrangement involves the correlation of the structure of macromolecules with their functions. A light scattering photometer was developed for detecting continuous measurement of the angular spectrum of light scattered by dynamically changing systems. The analysis of light scattered by biological macromolecules can be used to determine concentration, size, shape, molecular weight, and structural changes of cells, such as erythrocytes. Some light scattering photometers can collect and store 120 angular scattering spectra per minute, with an angular resolution of 0.2 deg which can be displayed with computer graphics. The light scattering photometer does the following: functions to produce and detect scattered light; determines scatter angles; and collects, stores, and analyzes data.

Johnson, Adriel D.↗

NASA NTRS1993

Microemulsion characterization by the use of a noninvasive backscatter fiber optic probe

This paper demonstrates the utility of a noninvasive backscatter fiber optic probe for dynamic light-scattering characterization of a microemulsion comprising sodium dodecyl sulfate/1-butanol/ brine/heptane. The fiber probe, comprising two optical fibers precisely positioned in a stainless steel body, is a miniaturized and efficient self-beating dynamic light-scattering system. Accuracy of particle size estimation is better than +/- 2 percent.

Ansari, Rafat R.↗

NASA NTRS1989

Laser light scattering as a probe of fractal colloid aggregates

The extensive use of laser light scattering is reviewed, both static and dynamic, in the study of colloid aggregation. Static light scattering enables the study of the fractal structure of the aggregates, while dynamic light scattering enables the study of aggregation kinetics. In addition, both techniques can be combined to demonstrate the universality of the aggregation process. Colloidal aggregates are now well understood and therefore represent an excellent experimental system to use in the study of the physical properties of fractal objects. However, the ultimate size of fractal aggregates is fundamentally limited by gravitational acceleration which will destroy the fractal structure as the size of the aggregates increases. This represents a great opportunity for spaceborne experimentation, where the reduced g will enable the growth of fractal structures of sufficient size for many interesting studies of their physical properties.

Weitz, David A.↗

NASA NTRS1996

Multiple-Fiber-Optic Probe For Light-Scattering Measurements

Multiple-fiber-optical probe developed for use in measuring light scattered at various angles from specimens of materials. Designed for both static and dynamic light-scattering measurements of colloidal dispersions. Probe compact, rugged unit containing no moving parts and remains stationary during operation. Not restricted to operation in controlled, research-laboratory environment. Positioned inside or outside light-scattering chamber. Provides simultaneous measurements at small angular intervals over range of angles, made to include small scattering angles by orienting probe in appropriate direction.

Dhadwal, Harbans Singh↗

NASA NTRS2002

Results From the Physics of Colloids Experiment on ISS

Weitz, David↗

NASA NTRS1998

The Dynamics of Disorder-Order Transition in Hard Sphere Colloidal Dispersions

The Physics of Hard Spheres Experiment (PHaSE) seeks a complete understanding of the entropically driven disorder-order transition in hard sphere colloidal dispersions. The light scattering instrument designed for flight collects Bragg and low angle light scattering in the forward direction via a CCD camera and performs conventional static and dynamic light scattering at 10-160 deg. through fiber optic cables. Here we report on the kinetics of nucleation and growth extracted from time-resolved Bragg images and measurements of the elastic modulus of crystalline phases obtained by monitoring resonant responses to sinusoidal forcing through dynamic light scattering. Preliminary analysis of the former indicates a significant difference from measurements on the ground, while the latter confirms nicely laboratory experiments with the same instrument and predictions from computer simulations.

Chaikin, Paul M.↗

NASA NTRS2002

PCS: The First Fluid Physics Payload on ISS

The Physics of Colloids in Space (PCS) experiment was accommodated within International Space Station (ISS) EXpedite the PRocessing of Experiments to Space Station (EXPRESS) Rack 2 and was remotely operated from early June 2001 until February 2002 from NASA Glenn Research Center's Telescience Support Center in Cleveland, Ohio and from a remote site at Harvard University in Cambridge, Massachusetts. PCS is an experiment conceived by Professor David A. Weitz of Harvard University (the Principal Investigator), focusing on the behavior of three different classes of colloid mixtures. The sophisticated light scattering instrumentation comprising PCS is capable of color imaging, and dynamic and static light scattering from 11 to 169 degrees, Bragg scattering over the range from 10 to 60 degrees, and laser light scattering at low angles from 0.3 to 6.0 degrees. The PCS instrumentation performed remarkably well, demonstrating a flexibility that enabled experiments to be performed that had not been envisioned prior to launch. While on-orbit, PCS accomplished 2400 hours of science operations, and was declared a resounding success. Each of the eight sample cells worked well and produced interesting and important results. Crystal nucleation and growth and the resulting structures of two binary colloidal crystal alloys were studied, with the long duration microgravity environment of the ISS facilitating extended studies on the growth and coarsening characteristics of the crystals. In another experiment run, the de-mixing of the colloid-polymer critical-point sample was studied as it phase-separates into two phases, one that resembles a gas and one that resembles a liquid. This process was studied over four decades of length scale, from 1 micron to 1 centimeter, behavior that cannot be observed in this sample on Earth because sedimentation would cause the colloids to fall to the bottom of the cell faster than the de-mixing process could occur. Similarly, the study of gelation and aging of another colloid-polymer sample, the colloid-polymer gel, also provided valuable information on gelation mechanisms, as did investigations on the extremely the low concentration silica and polystyrene fractal gel samples.

Doherty, M.↗