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A Fluorescence Recovery After Photobleaching (FRAP) Technique for the Measurement of Solute Transport Across Surfactant-Laden Interfaces

The technique of Fluorescence Recovery After Photobleaching (FRAP) has been applied to the measurement of interfacial transport in two-phase systems. FRAP exploits the loss of fluorescence exhibited by certain fluorophores when over-stimulated (photobleached), so that a two-phase system, originally at equilibrium, can be perturbed without disturbing the interface by strong light from an argon-ion laser and its recovery monitored by a microscope-mounted CCD camera as it relaxes to a new equilibrium. During this relaxation, the concentration profiles of the probe solute are measured on both sides of the interface as a function of time, yielding information about the transport characteristics of the system. To minimize the size of the meniscus between the two phases, a photolithography technique is used to selectively treat the glass walls of the cell in which the phases are contained. This allows concentration measurements to be made very close to the interface and increases the sensitivity of the FRAP technique.

Browne, Edward P.↗

Measurement of resistance to solute transport across surfactant-laden interfaces using a Fluorescence Recovery After Photobleaching (FRAP) technique

A noninvasive fluorescence recovery after photobleaching (FRAP) technique is under development to measure interfacial transport in two phase systems without disturbing the interface. The concentration profiles of a probe solute are measured in both sides of the interface by argon-ion laser, and the system relaxation is then monitored by a microscope-mounted CCD camera.

Browne, Edward P.↗

Photochemical and Microbial Degradation of Chromophoric Dissolved Organic Matter Exported from Tidal Marshes

Wetlands export chromophoric dissolved organic matter (CDOM) to estuaries, where CDOM is removed and transformed through biotic and abiotic process, subsequently impacting nutrient cycling, light availability, ecosystem metabolism, and phytoplankton activity. We examined the bioavailability and photoreactivity of CDOM exported from four Chesapeake Bay tidal marshes across three seasons and along an estuarine salinity gradient using three incubation treatments: 14-day microbial (MD), 7-day combined photochemical/microbial (PB+MD), and 7-day microbial incubation after photobleaching (MD after PB). CDOM absorption at 300 nm (aCDOM300) and dissolved organic carbon (DOC) concentrations showed strong seasonality, with minima in winter, but CDOM quality (absorption spectral slopes, fluorescence component ratios) was less variable seasonally. PB+MD over 7 days decreased aCDOM300 (–56.0%), humic-like fluorescence (–67.6%), and DOC (–17.8%), but increased the spectral slope ratio SR (=S275-295/S300-350) (+94.8%), suggesting a decrease in CDOM molecular weight. Photochemistry dominated the PB+MD treatment. Photoreactivity was greater during the winter and in marsh/watershed versus down-estuary sites, likely due to less previous light exposure. Prior photobleaching increased the bioavailability of marsh-exported CDOM, resulting in a greater loss of aCDOM300 and DOC, and a greater increase in humic-like fluorescence (–6.0%, –5.9%, and +18.4% change, respectively, over 7-day MD after PB incubations, versus –2.8%, –5.5%, and +2.6% change, respectively, over 14-day MD incubations). CDOM exported from a marsh downstream of a major wastewater treatment plant showed the greatest photoreactivity and bioavailability. This highlights the significance of human activity on estuarine CDOM quality and biogeochemical cycles.

Dissolved organic matter↗

Pulse energy dependence of subcellular dissection by femtosecond laser pulses

Precise dissection of cells with ultrashort laser pulses requires a clear understanding of how the onset and extent of ablation (i.e., the removal of material) depends on pulse energy. We carried out a systematic study of the energy dependence of the plasma-mediated ablation of fluorescently-labeled subcellular structures in the cytoskeleton and nuclei of fixed endothelial cells using femtosecond, near-infrared laser pulses focused through a high-numerical aperture objective lens (1.4 NA). We find that the energy threshold for photobleaching lies between 0.9 and 1.7 nJ. By comparing the changes in fluorescence with the actual material loss determined by electron microscopy, we find that the threshold for true material ablation is about 20% higher than the photobleaching threshold. This information makes it possible to use the fluorescence to determine the onset of true material ablation without resorting to electron microscopy. We confirm the precision of this technique by severing a single microtubule without disrupting the neighboring microtubules, less than 1 micrometer away. c2005 Optical Society of America.

Non-NASA Center↗

Estimating the Distribution of Colored Dissolved Organic Matter During the Southern Ocean Gas Exchange Experiment Using Four-Dimensional Variational Data Assimilation

We diagnosed the effect of various physical processes on the distribution of mixed-layer colored dissolved organic matter (CDOM) and a sulfur hexauoride (SF6) tracer during the Southern Ocean Gas Exchange Experiment (SO GasEx). The biochemical upper ocean state estimate uses in situ and satellite biochemical and physical data in the study region, including CDOM (absorption coefcient and spectral slope), SF6, hydrography, and sea level anomaly. Modules for photobleaching of CDOM and surface transport of SF6 were coupled with an ocean circulation model for this purpose. The observed spatial and temporal variations in CDOM were captured by the state estimate without including any new biological source term for CDOM, assuming it to be negligible over the 26 days of the state estimate. Thermocline entrainment and photobleaching acted to diminish the mixed-layer CDOM with time scales of 18 and 16 days, respectively. Lateral advection of CDOM played a dominant role and increased the mixed-layer CDOM with a time scale of 12 days, whereas lateral diffusion of CDOM was negligible. A Lagrangian view on the CDOM variability was demonstrated by using the SF6 as a weighting function to integrate the CDOM elds. This and similar data assimilation methods can be used to provide reasonable estimates of optical properties, and other physical parameters over the short-term duration of a research cruise, and help in the tracking of tracer releases in large-scale oceanographic experiments, and in oceanographic process studies.

Del Castillo, C. E.↗

Lunar dust and radiation darkening of Surveyor 3 surfaces

Measured spectral reflectance, evidence obtained from photographs, scanning electron microscopy, and previous work on Surveyor hardware were used to develop an understanding of the observed discoloration and its meaning to future space and lunar operations. Measured reflectance data were analyzed to separate and understand the effects of lunar dust and radiation damage. It was concluded that organic contamination is not a major contributor to the discoloration. Radiation-induced discoloration on the various surfaces was found to be proportional to the degree of solar illumination. Photobleaching of the radiation damage was observed and was responsible for a gradual change in the color of the camera's surface during the evaluation.

Carroll, W. F.↗

Annual Conference on Nuclear and Space Radiation Effects, 18th, University of Washington, Seattle, WA, July 21-24, 1981, Proceedings

Single event upset phenomena are discussed, taking into account cosmic ray induced errors in IIL microprocessors and logic devices, single event upsets in NMOS microprocessors, a prediction model for bipolar RAMs in a high energy ion/proton environment, the search for neutron-induced hard errors in VLSI structures, soft errors due to protons in the radiation belt, and the use of an ion microbeam to study single event upsets in microcircuits. Basic mechanisms in materials and devices are examined, giving attention to gamma induced noise in CCD's, the annealing of MOS capacitors, an analysis of photobleaching techniques for the radiation hardening of fiber optic data links, a hardened field insulator, the simulation of radiation damage in solids, and the manufacturing of radiation resistant optical fibers. Energy deposition and dosimetry is considered along with SGEMP/IEMP, radiation effects in devices, space radiation effects and spacecraft charging, EMP/SREMP, and aspects of fabrication, testing, and hardness assurance.

Tasca, D. M.↗

Nonlinear optical polymers for electro-optic signal processing

Photonics is an emerging technology, slated for rapid growth in communications systems, sensors, imagers, and computers. Its growth is driven by the need for speed, reliability, and low cost. New nonlinear polymeric materials will be a key technology in the new wave of photonics devices. Electron-conjubated polymeric materials offer large electro-optic figures of merit, ease of processing into films and fibers, ruggedness, low cost, and a plethora of design options. Several new broad classes of second-order nonlinear optical polymers were developed at the Navy's Michelson Laboratory at China Lake, California. Polar alignment in thin film waveguides was achieved by electric-field poling and Langmuir-Blodgett processing. Our polymers have high softening temperatures and good aging properties. While most of the films can be photobleached with ultraviolet (UV) light, some have excellent stability in the 500-1600 nm range, and UV stability in the 290-310 nm range. The optical nonlinear response of these polymers is subpicosecond. Electro-optic switches, frequency doublers, light modulators, and optical data storage media are some of the device applications anticipated for these polymers.

Lindsay, Geoffrey A.↗

Synthesis of phthalocyanine doped sol-gel materials

The synthesis of sol-gel silica materials doped with three different types of metallophthalocyanines has been studied. Homogeneous materials of good optical quality were prepared and the first optical limiting measurements of dyes in sol-gel hosts were carried out. The properties of these solid state limiters are similar to limiters based on phthalocyanine (Pc) in solution. Sol-gel silica materials containing copper, tin and germanium phthalocyanines were investigated. The initial step in all cases was to prepare silica sols by the sonogel method using tetramethoxy silane (TMOS), HCl and distilled water. Thereafter, the synthesis depended upon the specific Pc and its solubility characteristics. Copper phthalocyanine tetrasulfonic acid tetra sodium salt (CuPc4S) is soluble in water and various doping levels (1 x 10 (exp -4) M to 1 x 10 (exp -5) M) were added to the sol. The group IV Pc's, SnPc(OSi(n-hexyl)3)2 and GePc(OSi(n-hexyl)3)2, are insoluble in water and the process was changed accordingly. In these cases, the compounds were dissolved in THF and then added to the sol. The Pc concentration in the sol was 2 x 10(exp -5)M. The samples were then aged and dried in the standard method of making xerogel monoliths. Comparative nanosecond optical limiting experiments were performed on silica xerogels that were doped with the different metallophthalocyanines. The ratio of the net excited state absorption cross section (sigma(sub e)) to the ground state cross section (sigma(sub g)) is an important figure of merit that is used to characterize these materials. By this standard the SnPc sample exhibits the best limiting for the Pc doped sol-gel materials. Its cross section ratio of 19 compares favorably with the value of 22 that was measured in toluene. The GePc materials appear to not be as useful as those containing SnPc. The GePc doped solids exhibit a higher onset energy (2.5 mj and lower cross section ratio, 7. The CuPc4S sol-gel material has a still lower cross section ratio, 4, however, the tetrasulfonate groups make the dye soluble in water which greatly facilitates its incorporation into the sol-gel matrix. The nonlinear transmission of CuPc4S in a pH 2 buffer solution and in a silica xerogel were compared. It is evident that the CuPc4S preserves its optical limiting behavior in the sol-gel matrix, indicating that the fundamental excited state absorption process is essentially the same for a molecule in solution or in the solid state. Although the spectroscopic details of energy level lifetimes are unknown, the significance is that passive optical limiting has been achieved in the solid state via incorporation of a dye into an inorganic host. The only compromise occurs at the extremely high energy regime where photobleaching is observed. This is a result of the limited mobility of the dye molecules in the solid silica host relative to a liquid host. The effects of photodegradation in the xerogel are additive, whereas the solution provides a supply of fresh molecules that are free to enter the active volume between pulses.

Dunn, Bruce↗

Lightweight Fiber Optic Gas Sensor for Monitoring Regenerative Food Production

In this final report, Physical Optics Corporation (POC) describes its development of sensors for oxygen, carbon dioxide, and relative humidity. POC has constructed a phase fluorometer that can detect oxygen over the full concentration range from 0 percent to 100 percent. Phase-based measurements offer distinct advantages, such as immunity to source fluctuation, photobleaching, and leaching. All optics, optoelectronics, power supply, and the printed circuit board are included in a single box; the only external connections to the fluorometer are the optical fiber sensor and a power cord. The indicator-based carbon dioxide sensor is also suitable for short-term and discrete measurements over the concentration range from 0 percent to 100 percent. The optical fiber-based humidity sensor contains a porous core for direct interaction of the light beam with water vapor within fiber pores; the detection range for the humidity sensor is 10 percent to 100 percent, and response time is under five minutes. POC is currently pursuing the commercialization of these oxygen and carbon dioxide sensors for environmental applications.

Schmidlin, Edward↗

Flexible polyacrylamide substrata for the analysis of mechanical interactions at cell-substratum adhesions

We have described a powerful tool for the study of mechanical interactions between cells and their physical environment. Although the approach has already been used in a variety of ways to measure traction forces and to characterize active and passive responses of cultured cells to mechanical stimulation, it can be extended easily and combined with other microscopic approaches, including fluorescent analog imaging (Beningo et al., 2001), photobleaching, calcium imaging, micromanipulation, and electrophysiology. This method will be particularly useful for studying the functions of various components at focal adhesions, and the effects of mechanical forces on focal adhesion-mediated signal transduction. In addition, the method can be extended to a 3D setting, e.g., by sandwiching cultured cells between two layers of polyacrylamide to create an environment mimicking that in the tissue of a multicellular organism. Whereas chemical interactions between cells and the environment have been investigated extensively, many important questions remain as to the role of physical forces in cellular functions and the interplay between chemical and physical mechanisms of communication. The present approach, as well as other approaches capable of probing physical interactions, should fill in this important gap in the near future.

NASA Discipline Cell Biology↗

Protein sequences and redox titrations indicate that the electron acceptors in reaction centers from heliobacteria are similar to Photosystem I

Photosynthetic reaction centers isolated from Heliobacillus mobilis exhibit a single major protein on SDS-PAGE of 47 000 Mr. Attempts to sequence the reaction center polypeptide indicated that the N-terminus is blocked. After enzymatic and chemical cleavage, four peptide fragments were sequenced from the Heliobacillus mobilis apoprotein. Only one of these sequences showed significant specific similarity to any of the protein and deduced protein sequences in the GenBank data base. This fragment is identical with 56% of the residues, including both cysteines, found in highly conserved region that is proposed to bind iron-sulfur center Fx in the Photosystem I reaction center peptide that is the psaB gene product. The similarity to the psaA gene product in this region is 48%. Redox titrations of laser-flash-induced photobleaching with millisecond decay kinetics on isolated reaction centers from Heliobacterium gestii indicate a midpoint potential of -414 mV with n = 2 titration behavior. In membranes, the behavior is intermediate between n = 1 and n = 2, and the apparent midpoint potential is -444 mV. This is compared to the behavior in Photosystem I, where the intermediate electron acceptor A1, thought to be a phylloquinone molecule, has been proposed to undergo a double reduction at low redox potentials in the presence of viologen redox mediators. These results strongly suggest that the acceptor side electron transfer system in reaction centers from heliobacteria is indeed analogous to that found in Photosystem I. The sequence similarities indicate that the divergence of the heliobacteria from the Photosystem I line occurred before the gene duplication and subsequent divergence that lead to the heterodimeric protein core of the Photosystem I reaction center.

NASA Discipline Number 52-30↗

Organic materials able to detect analytes

The present invention generally relates to polymers with lasing characteristics that allow the polymers to be useful in detecting analytes. In one aspect, the polymer, upon an interaction with an analyte, may exhibit a change in a lasing characteristic that can be determined in some fashion. For example, interaction of an analyte with the polymer may affect the ability of the polymer to reach an excited state that allows stimulated emission of photons to occur, which may be determined, thereby determining the analyte. In another aspect, the polymer, upon interaction with an analyte, may exhibit a change in stimulated emission that is at least 10 times greater with respect to a change in the spontaneous emission of the polymer upon interaction with the analyte. The polymer may be a conjugated polymer in some cases. In one set of embodiments, the polymer includes one or more hydrocarbon side chains, which may be parallel to the polymer backbone in some instances. In another set of embodiments, the polymer may include one or more pendant aromatic rings. In yet another set of embodiments, the polymer may be substantially encapsulated in a hydrocarbon. In still another set of embodiments, the polymer may be substantially resistant to photobleaching. In certain aspects, the polymer may be useful in the detection of explosive agents, such as 2,4,6-trinitrotoluene (TNT) and 2,4-dinitrotoluene (DNT).

Rose, Aimee↗