Method for separating biological cells

A method for separating biological cells by suspending a mixed cell population in a two-phase polymer system is described. The polymer system consists of droplet phases with different surface potentials for which the cell populations exhibit different affinities. The system is subjected to an electrostatic field of sufficient intensity to cause migration of the droplets with an attendant separation of cells

Cell partition in two phase polymer systems

Aqueous phase-separated polymer solutions can be used as support media for the partition of biological macromolecules, organelles and cells. Cell separations using the technique have proven to be extremely sensitive to cell surface properties but application of the systems are limited to cells or aggregates which do not significantly while the phases are settling. Partition in zero g in principle removes this limitation but an external driving force must be applied to induce the phases to separate since their density difference disappears. We have recently shown that an applied electric field can supply the necessary driving force. We are proposing to utilize the NASA FES to study field-driven phase separation and cell partition on the ground and in zero g to help define the separation/partition process, with the ultimate goal being to develop partition as a zero g cell separation technique

Countercurrent distribution of biological cells

A neutral polymer phase system consisting of 7.5 percent dextran 40/4.5 percent PEG 6, 0.11 M Na phosphate, 5 percent fetal bovine serum (FBS), pH 7.5, was developed which has a high phase droplet electrophoretic mobility and retains cell viability over many hours. In this and related systems, the drop mobility was a linear function of drop size, at least in the range 4-30 micron diameter. Applications of and electric field of 4.5 v/cm to a system containing 10 percent v/v bottom phase cleared the system more than two orders of magnitude faster than in the absence of the field. At higher bottom phase concentrations a secondary phenomenon intervened in the field driven separations which resulted in an increase in turbidity after clearing had commenced. The increase was associated with a dilution of the phase system in the chamber. The effect depended on the presence of the electric field. It may be due to electroosmotic flow of buffer through the Amicon membranes into the sample chamber and flow of phase system out into the rinse stream. Strategies to eliminate this problem are proposed

SPAR project: Counter current distribution of biologicals

Preliminary ground based experiments strongly support the validity of a proposed space experiment in the separation of biological cells by counter current distribution (CCD). It was shown that a low level applied selective field can accelerate the separation of the phases by a very large factor. An electric field therefore should be very effective in space in lieu of gravity for the phase separation step. This appears to solve a central problem in implementing CCD in space and opens the technique to many types of large cell mixtures for which it has not been applicable before due to rapid settling effects. It was determined that electrophoretic migration of the cells, as a possibly disturbing factor, will be of negligible effect. Attenuation of a light beam by scattering generated a signal indicating the course of the phase separation

Study of ephemeris accuracy of the minor planets

The current state of minor planet ephemerides was assessed, and the means for providing and updating these emphemerides for use by both the mission planner and the astronomer were developed. A system of obtaining data for all the numbered minor planets was planned, and computer programs for its initial mechanization were developed. The computer based system furnishes the osculating elements for all of the numbered minor planets at an adopted date of October 10, 1972, and at every 400 day interval over the years of interest. It also furnishes the perturbations in the rectangular coordinates relative to the osculating elements at every 4 day interval. Another computer program was designed and developed to integrate the perturbed motion of a group of 50 minor planets simultaneously. Sampled data resulting from the operation of the computer based systems are presented

Basic hydrogeologic and remote sensing data for selection of sanitary landfill sites

Solid waste disposal were studied in Volusia County to protect the water supply in the area. Highlands in this County are of limited areal extent and, most significantly, the sand hills and ridges are in areas where recharge of the Floridan aquifer occurs. This study proves that well drained soils meeting the current State requirements are of limited areal extent. These areas should not be utilized as sanitary landfill sites! Rather, it is recommended that the Tomoka Farm Road site into the adjacent wetlands be extended. The County site on Rima Ridge recommended by Greenleaf-Telesca as the primary waste burial site in the County should be re-evaluated because of potential danger to the Daytona Beach water supply

Method for detecting coliform organisms

A method and apparatus are disclosed for determining the concentration of coliform bacteria in a sample. The sample containing the coliform bacteria is cultured in a liquid growth medium. The cultured bacteria produce hydrogen and the hydrogen is vented to a second cell containing a buffer solution in which the hydrogen dissolves. By measuring the potential change in the buffer solution caused by the hydrogen, as a function of time, the initial concentration of bacteria in the sample is determined. Alternatively, the potential change in the buffer solution can be compared with the potential change in the liquid growth medium to verify that the potential change in the liquid growth medium is produced primarily by the hydrogen gas produced by the coliform bacteria

Countercurrent distribution of biological cells

Detailed physiochemical studies of dextran/poly(ethylene glycol) (PEG) two phase systems were carried out to characterize and provide understanding of the properties of the systems which determine cell partition and the electrophoretic behavior of phase drops responsible for electric field driven phase separation. A detailed study of the electrostatic and electrokinetic potentials developed in these systems was carried out. The salt partition was examined both in phase systems and with pure polymer solutions via equilibrium dialysis and mechanism of sulfate, chloride and phosphate partition shown to be exclusion by PEG rather than binding by dextran. Salt partition was shown to have a strong effect on the polymer compositions of the phases as well, an effect which produces large changes in the interfacial tension between them. These effects were characterized and the interfacial tension shown to obey a power law with respect to its dependence on the length of the tie line describing the system composition on a phase diagram. The electrostatic potential differences measured via salt bridges were shown to obey thermodynamic predictions. The electrophoretic mobilities measured were utilized to provide a partial test of Levine's incomplete theory of phase drop electrophoresis. The data were consistent with Levine's expression over a limited range of the variables tested