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At least 163 records · Page 9

Analysis of electrophoresis performance

A flexible efficient computer code is being developed to simulate electrophoretic separation phenomena, in either a cylindrical or a rectangular geometry. The code will computer the evolution in time of the concentrations of an arbitrary number of chemical species, and of the temperature, pH distribution, conductivity, electric field, and fluid motion. Use of nonuniform meshes and fast accurate implicit time-stepping will yield accurate answers at economical cost.

Roberts, Glyn O.↗

Electrophoresis and isoelectric focusing of whole cell and membrane proteins from the extremely halophilic archaebacteria

The subunits from two purified halobacterial membrane enzymes (ATPase and nitrate reductase) behaved differently with respect to isoelectric focusing, silver staining and interaction with ampholytes. Differential behavior was also observed in whole cell proteins from Halobacterium saccharovorum regarding resolution in two-dimensional gels and silver staining. It is proposed that these differences reflect the existence of two classes of halobacterial proteins.

Stan-Lotter, Helga↗

Application of partition technology to particle electrophoresis

The effects of polymer-ligand concentration on particle electrophoretic mobility and partition in aqueous polymer two-phase systems are investigated. Polymer coating chemistry and affinity ligand synthesis, purification, and analysis are conducted. It is observed that poly (ethylene glycol)-ligands are effective for controlling particle electrophoretic mobility.

Van Alstine, James M.↗

Nanodiamond in Colloidal Suspension: Electrophoresis; Other Observations

Selective laser extraction has demonstrated that meteoritic diamonds may consist of subpopulations with different optical absorption properties, but it is not clear what makes them optically different. More work is needed to understand the mechanism for selective laser extraction. Additional information is contained in the original extended abstract.

Meshik, A. P.↗

Microchip Non-Aqueous Capillary Electrophoresis (MicronNACE) Method to Analyze Long-Chain Primary Amines

A protocol was developed as a first step in analyzing the complex organic aerosols present on Saturn's moon Titan, as well as the analogues of these aerosols (tholins) made on Earth. Labeling of primary amines using Pacific Blue succinimidyl ester is effected in ethanol with 25 mM triethylamine to maintain basic conditions. This reaction is allowed to equilibrate for at least one hour. Separation of the labeled primary amines is performed in ethanol with 1.05 M acetic acid, and 50 mM ammonium acetate in a commercial two-layer glass device with a standard crossmicrochannel measuring 50 microns wide by 20 microns deep. Injection potentials are optimized at 2 kV from the sample (negative) to the waste well (positive), with slight bias applied to the other two wells ( 0.4 and 0.8 V) to pinch the injection plug for the 30-s injection. Separation is performed at a potential of 5 kV along the channel, which has an effective separation distance of 7 cm. The use of ethanol in this method means that long-chain primary amines can be dissolved. Due to the low pH of the separation buffer, electro-osmotic flow (EOF) is minimized to allow for separation of both short-chain and longchain amines. As the freezing point of ethanol is much lower than water, this protocol can perform separations at temperatures lower than 0 C, which would not be possible in aqueous phase. This is of particular importance when considering in situ sampling of Titan aerosols, where unnecessary heating of the sample (even to room temperature) would lead to decomposition or unpredictable side reactions, which would make it difficult to characterize the sample appropriately.

Willis, Peter A.↗

Microfabricated Genomic Analysis System

Genetic sequencing and many genetic tests and assays require electrophoretic separation of DNA. In this technique, DNA fragments are separated by size as they migrate through a sieving gel under the influence of an applied electric field. In order to conduct these analyses on-orbit, it is essential to acquire the capability to efficiently perform electrophoresis in a microgravity environment. Conventional bench top electrophoresis equipment is large and cumbersome and does not lead itself to on-orbit utilization. Much of the previous research regarding on-orbit electrophoresis involved altering conventional electrophoresis equipment for bioprocessing, purification, and/or separation technology applications. A new and more efficient approach to on-orbit electrophoresis is the use of a microfabricated electrophoresis platform. These platforms are much smaller, less expensive to produce and operate, use less power, require smaller sample sizes (nanoliters), and achieve separation in a much shorter distance (a few centimeters instead of 10 s or 100 s of centimeters.) In contrast to previous applications, this platform would be utilized as an analytical tool for life science/medical research, environmental monitoring, and medical diagnoses. Identification of infectious agents as well as radiation related damage are significant to NASA s efforts to maintain, study, and monitor crew health during and in support of near-Earth and interplanetary missions. The capability to perform genetic assays on-orbit is imperative to conduct relevant and insightful biological and medical research, as well as continuing NASA s search for life elsewhere. This technology would provide an essential analytical tool for research conducted in a microgravity environment (Shuttle, ISS, long duration/interplanetary missions.) In addition, this technology could serve as a critical and invaluable component of a biosentinel system to monitor space environment genotoxic insults to include radiation.

Gonda, Steve↗

Moving wall, continuous flow electronphoresis apparatus

This invention relates generally to electrophoresis devices and more particularly to a moving wall, continuous flow device in which an electrophoresis chamber is angularly positionable with respect to the direction of moving belt walls. A frame with an electrophoresis chamber is rotatably supported between two synchronously driven belt walls. This allows the chamber to be angularly positionable with respect to the direction of belt travel, which compensates for electroosmotic flow within the electrophoresis chamber. Injection of a buffer solution via an opening and a homogenous sample stream via another opening is performed at the end of a chamber, and collection of buffer and the fractionated species particles is done by a conventional collection array at an opposite end of the chamber. Belts are driven at a rate which exactly matches the flow of buffer and sample through the chamber, which entrains the buffer to behave as a rigid electrophoretic medium, eliminating flow distortions (Poiseuille effect). Additionally, belt material for each belt is stored at one end of the device and is taken up by drive wheels at an opposite end. The novelty of this invention particularly lies in the electrophoresis chamber being angularly positionable between two moving belt walls in order to compensate for electroosmotic flow. Additionally, new belt material is continuously exposed within the chamber, minimizing flow distortion due to contamination of the belt material by the sample.

Rhodes, Percy H.↗

Visualization of yeast chromosomal DNA

The DNA molecule is the most significant life molecule since it codes the blue print for other structural and functional molecules of all living organisms. Agarose gel electrophoresis is now being widely used to separate DNA of virus, bacteria, and lower eukaryotes. The task was undertaken of reviewing the existing methods of DNA fractionation and microscopic visualization of individual chromosonal DNA molecules by gel electrophoresis as a basis for a proposed study to investigate the feasibility of separating DNA molecules in free fluids as an alternative to gel electrophoresis. Various techniques were studied. On the molecular level, agarose gel electrophoresis is being widely used to separate chromosomal DNA according to molecular weight. Carl and Olson separate and characterized the entire karyotype of a lab strain of Saccharomyces cerevisiae. Smith et al. and Schwartz and Koval independently reported the visualization of individual DNA molecules migrating through agarose gel matrix during electrophoresis. The techniques used by these researchers are being reviewed in the lab as a basis for the proposed studies.

Lubega, Seth↗

Analytical study of electrophoretic characterization of kidney cells

Embryonic kidney cells were studied as a follow-up to the MA-011 Electrophoresis Technology Experiment which was conducted during the Apollo Soyuz Test Project (ASTP). The postflight analysis of the performance of the ASTP zone electrophoresis experiment involving embryonic kidney cells is reported. The feasibility of producing standard particles for electrophoresis was also studied. This work was undertaken in response to a need for standardization of methods for producing, calibrating, and storing electrophoretic particle standards which could be employed in performance tests of various types of electrophoresis equipment. Promising procedures were tested for their suitability in the production of standard test particles from red blood cells.

Knox, R. J.↗

Multistage Electrophoretic Separators

A multistage electrophoresis apparatus has been invented for use in the separation of cells, protein molecules, and other particles and solutes in concentrated aqueous solutions and suspensions. The design exploits free electrophoresis but overcomes the deficiencies of prior free-electrophoretic separators by incorporating a combination of published advances in mathematical modeling of convection, sedimentation, electro-osmotic flow, and the sedimentation and aggregation of droplets. In comparison with other electrophoretic separators, these apparatuses are easier to use and are better suited to separation in relatively large quantities characterized in the art as preparative (in contradistinction to smaller quantities characterized in the art as analytical). In a multistage electrophoretic separator according to the invention, an applied vertical steady electric field draws the electrically charged particles of interest from within a cuvette to within a collection cavity that has been moved into position of the cuvette. There are multiple collection cavities arranged in a circle; each is aligned with the cuvette for a prescribed short time. The multistage, short-migration-path character of the invention solves, possibly for the first time, the fluid-instability problems associated with free electrophoresis. The figure shows a prototype multistage electrophoretic separator that includes four sample stations and five collection stages per sample. At each sample station, an aqueous solution or suspension containing charged species to be separated is loaded into a cuvette, which is machined into a top plate. The apparatus includes a lower plate, into which 20 collection cavities have been milled. Each cavity is filled with an electrophoresis buffer solution. For the collection of an electrophoretic fraction, the lower plate is rotated to move a designated collection cavity into alignment with the opening of the cuvette. An electric field is then applied between a non-gassing electrode in the collection cavity and an electrolyte compartment, which is separated from the cuvette by a semipermeable membrane. The electrolyte is refreshed by circulation by use of a peristaltic pump. In subsequent steps, the lower plate is rotated to collect other electrophoretic fractions. Later, the collected fractions are removed from the collection cavities through ports that have threaded plugs. The base of the apparatus contains power supplies and a computer interface. The design includes provisions for monitoring and feedback control of cavity position, electric field, and temperature. The operation of the apparatus can easily be automated, as demonstrated by use of software that has already been written for this purpose.

Thomas, Nathan↗

Electrophoretic characterization of aldehyde-fixed red blood cells, kidney cells, lynphocytes and chamber coatings

Ground-based electrokinetic data on the electrophoresis flight experiment to be flown on the Apollo-Soyuz Test Project experiment MA-011 are stipulated. Aldehyde-fixed red blood cells, embryonic kidney cells and lymphocytes were evaluated by analytical particle electrophoresis. The results which aided in the interpretation of the final analysis of the MA-011 experiment are documented. The electrophoresis chamber surface modifications, the buffer, and the material used in the column system are also discussed.

Source record↗

Investigation of the free flow electrophoretic process. Volume 2: Technical analysis

The effect of gravity on the free flow electrophoretic process was investigated. The demonstrated effects were then compared with predictions made by mathematical models. Results show that the carrier buffer flow was affected by gravity induced thermal convection and that the movement of the separating particle streams was affected by gravity induced buoyant forces. It was determined that if gravity induced buoyant forces were included in the mathematical models, then effective predictions of electrophoresis chamber separation performance were possible. The results of tests performed using various methods of electrophoresis using supportive media show that the mobility and the ability to separate were essentially independent of concentration, providing promise of being able to perform electrophoresis with higher inlet concentrations in space.

Weiss, R. A.↗

Electrophoretic purification of cells in space - Evaluation of results from STS-3

The procedure and results of Electrophoresis Equipment Verification Test, designed to examine electrophoretic behavior of animal cells is suspension more concentrated than possible on earth and flown on the Shuttle flight STS-3, were discussed. Ground-based laboratory values of electrophoretic mobilities of a mixture of human and rabbit aldehyde-fixed red blood cells (RBC) were compared with those recorded at 11 minute intervals on the Shuttle STS-3. RBC migration and separation observed through photographic records were not as expected. However, cell mobilities and migrating band profiles were consistent with the results of laboratory simulation experiments. It was concluded that zero G electrophoresis of very high concentrations (1 x 10 to the 9th) is possible and similar to electrophoresis of normal cell concentrations on earth.

Sarnoff, B. E.↗

Advances in electrophoretic separations

Free fluid electrophoresis is described using laboratory and space experiments combined with extensive mathematical modeling. Buoyancy driven convective flows due to thermal and concentration gradients are absent in the reduced gravity environment of space. The elimination of convection in weightlessness offers possible improvements in electrophoresis and other separation methods which occur in fluid media. The mathematical modeling suggests new ways of doing electrophoresis in space and explains various phenomena observed during past experiments. The extent to which ground based separation techniques are limited by gravity induced convection is investigated and space experiments are designed to evaluate specific characteristics of the fluid/particle environment. A series of experiments are proposed that require weightlessness and apparatus is developed that can be used to carry out these experiments in the near future.

Snyder, R. S.↗