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PubMed2005

Combination of gravitational SPLITT fractionation and field-flow fractionation for size-sorting and characterization of sea sediment.

A combination of gravitational split-flow thin (SPLITT) fractionation and sedimentation/steric field-flow fractionation (Sd/StFFF) has been used for continuous size-sorting of a sediment sample and for size analysis of the collected fractions. An IAEA (International Atomic Energy Agency) sediment material was separated into four size fractions (with theoretical size ranges <1.0, 1.0-3.0, 3.0-5.0, and >5.0 microm in diameter) by means of a three-step gravitational SPLITT fractionation (GSF) for which the same GSF channel was used throughout. The GSF fractions were collected and examined by optical microscopy (OM) and by Sd/St FFF. The mean diameters of the GSF fractions measured by OM were within the size interval predicted by GSF theory, despite the theory assuming that all particles are spherical, which is not true for the sediment particles. The Sd/St FFF results showed that retention shifted toward shorter elution time (or larger size) than expected, probably because of the shape effect. The results from GSF, OM, and Sd/StFFF are discussed in detail.

Calibration↗
PubMed1999

Field- and flow-dependent trapping of red blood cells on polycarbonate accumulation wall in sedimentation field-flow fractionation.

Sedimentation field-flow fractionation (SdFFF) instrumentation is now mature. Methodological procedure and particle separation development rules are well established even in the case of biological species. However, in some biological applications, retention properties of samples not predicted by any field-flow fractionation (FFF) elution models are observed. It is demonstrated that the trapping of cellular material in the separation system is not related to geometrical instrumentation features but to channel wall characteristics. The physicochemical particle-wall attractive interactions are different depending on the flow-rate and field intensity applied. Separation power in SdFFF for biological species is therefore limited by the intensity of these interactions. In terms of separation, a balance is to be found between external field and flow intensity to limit particle-wall interactions.

Cell Separation↗
PubMed2006

Electric circuit model for electrical field flow fractionation.

In electrical field flow fractionation (EFFF or ElFFF), an electric potential is applied across a narrow gap filled with a weak electrolyte fluid. Charge buildup at the two poles (electrodes) and the formation of an electric double layer shields the channel, making the effective field in the bulk fluid very weak. Recent computational research suggests that pulsed field protocols, however, should improve retention and may enhance separation in EFFF through systematic disruptions of the double layer resulting in a stronger effective field in the bulk fluid. Improved retention has already been demonstrated experimentally. Accurate modeling and subsequent device optimization and design, however, depends, in part, on formulating a suitable model for the capacitative response of the channel and double layer at the electrode surfaces. Early models do not correctly describe experimentally observed current-time response and are not physically meaningful even when accurate mathematical fits of the data are realized. A new model and conceptual framework based on electrical resistance and capacitance variations of the double layer is suggested here. Physical interpretations of the electrical response have been developed and compared to published experimental data sets.

Electrochemistry↗
PubMed2005

Characterization of surface-modified nanoparticles for in vivo biointeraction. A sedimentation field flow fractionation study.

Sedimentation field flow fractionation (SdFFF) is an emerging high-performance analytical tool for separation and determination of size and adsorption characteristics of colloidal particles. This study demonstrates how SdFFF can be used to characterize nanoparticles prepared for in vivo applications including (1) the quantification of polymer uptake on nanoparticles where surface coverage is crucial and (2) the coupling of cell adhesive peptides containing the Arg-Gly-Asp motif (RGD). Quantitative information about polymer adhesion in order to prepare a bioinert surface and an accurate determination of ligand uptake are both of obvious importance for the understanding of, for example, relations between the number of attached molecules for biointeraction and an observed therapeutic effect. In addition, the present work highlights the necessity to perform careful characterization of commercially available particulate starting materials, in terms of size and polydispersity, prior to biological experimentation.

Fractionation, Field Flow↗
PubMed2001

Size- and shape-dependent separation of TiO2 colloidal sub-populations with gravitational field flow fractionation.

The simplest field flow fractionation technique, which uses the earth's gravity as the external field is applied to isolate two populations, which differ in both shape and size, from a polydisperse sub-micron TiO2 powder of homogenous density. The fraction eluted first is spherical with an average diameter of 0.31 microm while the second fraction is ellipsoidal and can be associated with a 0.45 microm hydrodynamic diameter. Elution conditions appeared to be very sensitive to electrolyte and surfactant characteristics in the carrier phase as well as on the sample concentration. Using 25 microl (1%, w/w) sample suspension, separations of spherical from ovoid particles was performed in almost 2 h with a mobile phase of 0.001 M KNO3-0.01% (v/v) Fl-70 in water in a 0.025-cm thick channel made of polystyrene walls.

Chemical Fractionation↗
PubMed1990

Two-dimensional field-flow fractionation.

Multidimensional field-flow fractionation (FFF) is described in two major forms: one in which different separative stages are coupled together and one in which two independent displacements, at least one of them FFF, are carried out in a generally planar channel structure. The latter, the subject of this paper, is relatively promising for implementation in FFF systems because in most cases the geometry of the FFF channel is already planar; the channel structure needs mainly to be broadened along the second dimension and modified with different inlets and outlets for this two-dimensional use. The large number of potential two-dimensional FFF systems is discussed. These systems are described at greater length in four categories: (1) FFF displacement used in both dimensions, (2) FFF along one axis and chromatography along another, (3) FFF along one axis and a field-induced displacement along the other, and (4) FFF separation combined with bulk or flow displacement at right angles. Finally, theoretical equations are obtained for the deflection of the trajectories away from the main flow axis z. The sensitivity of deflection to component properties is described in terms of the deflection selectivity. Several examples are discussed in which the deflection selectivity is remarkably high.

Chemical Fractionation↗
PubMed1999

Cell separation on microfabricated electrodes using dielectrophoretic/gravitational field-flow fractionation.

Dielectrophoretic/gravitational field-flow fractionation (DEP/G-FFF) was used to separate cultured human breast cancer MDA-435 cells from normal blood cells mixed together in a sucrose/dextrose medium. An array of microfabricated, interdigitated electrodes of 50 microns widths and spacings, and lining the bottom surface of a thin chamber (0.42 mm H x 25 mm W x 300 mm L), was used to generate DEP forces that levitated the cells. A 10-microL cell mixture sample containing approximately 50,000 cells was introduced into the chamber, and cancerous and normal blood cells were levitated to different heights according to the balance of DEP and gravitational forces. The cells at different heights were transported at different velocities under the influence of a parabolic flow profile that was established in the chamber and were thereby separated. Separation performance depended on the frequency and voltage of the applied DEP field and the fluid-flow rate. It took as little as 5 min to achieve cell separation. An analysis of the DEP/G-FFF results revealed that the separation exploited the difference in dielectric and density properties between cell populations. The DEP/G-FFF technique is potentially applicable to many biological and biomedical problems, especially those related to microfluidic systems.

Algorithms↗
PubMed2000

Development of electrical field-flow fractionation

Electrical field-flow fractionation (ElFFF) results for a series of polystyrene latex beads are presented. To first approximation, retention behavior can be related to conventional FFF theory, modified to account for a particle-wall repulsion effect. Size selectivity and column efficiency were exceptionally high, again approaching the upper limit predicted by theory. For the channel described in the present study, application of small voltages (typically less than 2 V) across the thin (131 microm) separation space defined by a Teflon spacer generates nominal field strengths of 10(4) V m(-1). However, electrode polarization reduces the effective field across the bulk of the channel to approximately 3% of the nominal value in the system studied. The magnitude of the applied field was calibrated by using standard latex beads of known size and mobility. Perturbations to retention behavior, such as overloading, were investigated. It was found that ideal separations occur at very dilute concentrations of the sample plug and that working in systems of very low ionic strength, the double-layer thickness adds significantly to the effective size of a particle. Steric inversion was observed at a particle size of approximately 0.4 microm under the conditions employed.

Journal Article↗
PubMed1981

Characterization of albumin microspheres by sedimentation field-flow fractionation.

Sedimentation field-flow fractionation (FFF) is a new technique that separates and characterizes submicron particles. In the present work, two independent sedimentation FFF methods are presented to characterize bovine serum albumin microspheres in terms of particle size, polydispersity, and diffusion coefficient. Particle diameters and polydispersities determined by the two sedimentation FFF methods were in excellent agreement with each other and in good agreement with values calculated from transmission electron microscopy (TEM) measurements. The diameters calculated from the two FFF methods and TEM were 0.349, 0.346, and 0.354 microgram, respectively.

Chemical Fractionation↗
PubMed2006

Observation of salt-induced beta-lactoglobulin aggregation using sedimentation field-flow fractionation.

Sedimentation field-flow fractionation (SdFFF) was applied in order to characterize particle sizes of beta-lactoglobulin aggregates induced by Ca2+ or Zn2+. Aggregation induced by Zn2+ was faster than that induced by Ca2+. Effects of Zn2+ and beta-lactoglobulin concentrations, as well as contact time, on the aggregation of beta-lactoglobulin were examined. All factors exhibited a combined effect on the size of aggregates, whereby larger aggregates were obtained at increased concentrations of Zn2+ and beta-lactoglobulin. At fixed concentrations of 2% (w/v) beta-lactoglobulin and 10 mM Zn2+, the particle size of the aggregates increased from 0.19 microm (at 15 min) to 0.38 microm (at 2880 min). Further, a hyphenated technique of SdFFF and inductively coupled plasma-optical emission spectrometry (ICP-OES) was used to examine whether intermolecular ionic bridges take part in salt-induced beta-lactoglobulin aggregation. With SdFFF-ICP-OES, protein-cation-protein cross-linkages were observed for beta-lactoglobulin aggregation induced by Zn2+, but not for that induced by Ca2+.

Calcium↗
PubMed1995

Selective elution and purification of living Trichomonas vaginalis using gravitational field-flow fractionation.

Gravitational field-flow fractionation is one of the simplest separation methods for biological materials. Its potential in parasitology is demonstrated for Trichomonas vaginalis, a parasite responsible for one of the most widespread sexually transmitted diseases. It was observed that this unicellular parasite can be purified in a culture medium with a recovery of 85% for the living trophozoites. The parasite retention characteristics were different when motile living and non-motile dead cells were eluted, motile cells being less retained than the non-motile cells.

Adult↗
PubMed1996

Micropreparation of hemopoietic stem cells from the mouse bone marrow suspension by gravitational field-flow fractionation.

Gravitational field-flow fractionation is a relatively simple experimental technique. This method was used for the characterization of stem cells from mouse bone marrow. Because these cells are bigger than the other cells in bone marrow, it is possible to separate them from the mixture. The fractions collected after passing through the separation channel were characterized using a Coulter Counter and used for transplantation into irradiated mice.

Animals↗
PubMed2000

Cell separation by dielectrophoretic field-flow-fractionation.

Dielectrophoretic field-flow-fractionation (DEP-FFF) was applied to several clinically relevant cell separation problems, including the purging of human breast cancer cells from normal T-lymphocytes and from CD34+ hematopoietic stem cells, the separation of the major leukocyte subpopulations, and the enrichment of leukocytes from blood. Cell separations were achieved in a thin chamber equipped with a microfabricated, interdigitated electrode array on its bottom wall that was energized with AC electric signals. Cells were levitated by the balance between DEP and sedimentation forces to different equilibrium heights and were transported at differing velocities and thereby separated when a velocity profile was established in the chamber. This bulk-separation technique adds cell intrinsic dielectric properties to the catalog of physical characteristics that can be applied to cell discrimination. The separation process and performance can be controlled through electronic means. Cell labeling is unnecessary, and separated cells may be cultured and further analyzed. It can be scaled up for routine laboratory cell separation or implemented on a miniaturized scale.

Animals↗
PubMed1999

The removal of human breast cancer cells from hematopoietic CD34+ stem cells by dielectrophoretic field-flow-fractionation.

Dielectrophoretic field-flow-fractionation (DEP-FFF) was used to purge human breast cancer MDA-435 cells from hematopoietic CD34+ stem cells. An array of interdigitated microelectrodes lining the bottom surface of a thin chamber was used to generate dielectrophoretic forces that levitated the cell mixture in a fluid flow profile. CD34+ stem cells were levitated higher, were carried faster by the fluid flow, and exited the separation chamber earlier than the cancer cells. Using on-line flow cytometry, efficient separation of the cell mixture was observed in less than 12 min, and CD34+ stem cell fractions with a purity >99.2% were obtained. The method of DEP-FFF is potentially applicable to many biomedical cell separation problems, including microfluidic-scale diagnosis and preparative-scale purification of cell subpopulations.

Bone Marrow Purging↗
PubMed2003

Size-based analysis of incinerator fly ash using gravitational SPLITT fractionation, sedimentation field-flow fractionation, and inductively coupled plasma-atomic emission spectroscopy.

Fly ash has been regarded as hazardous because of its high adsorption of toxic organic and/or inorganic pollutants. Fly ash is also known to have broad distributions of different chemical and physical properties, such as size and density. In this study, fly ash emitted from a solid waste incinerator was pre-fractionated into six sub-populations by use of gravitational SPLITT fractionation (GSF). The GSF fractions were then analyzed by sedimentation field-flow fractionation (SdFFF) and ICP-AES. SdFFF analysis showed the fly ash has a broad size distribution ranging from a few nanometers up to about 50 microm. SdFFF results were confirmed by electron microscopy. Inductively coupled plasma-atomic emission spectroscopy (ICP-AES) analysis of the GSF fractions showed the fly-ash particles contain a variety of inorganic elements including Ca, Si, Mg, Fe, and Pb. The most abundant in fly ash was Ca, followed by Si then Mg. No correlations were found between trace element concentration and particle size.

Carbon↗
PubMed1976

Flow-field-flow fractionation: a versatile new separation method.

Flow-field-fractionation is a new separation technique that seems likely to have broad applicability. Its theoretical scope includes any solute for which one can find a solvent and a semipermeable membrane. The principles on which the technique is based are presented. Some experiments with polystyrene beads, viruses, and proteins verify that retention depends solely on diffusion coefficients.

Chemical Fractionation↗
PubMed2005

Comparison of centrifugation and filtration techniques for the size fractionation of colloidal material in soil suspensions using sedimentation field-flow fractionation.

Sedimentation field-flow fractionation (SdFFF) with UV detection is used to systematically investigate the effect of traditional membrane filtration and centrifugation procedures on the isolation of specific size fractions from soil suspensions. Both procedures were used to isolate the nominal <0.45 and <0.2 microm fractions from a clay soil suspension. Results showed that the membrane filtration approach seriously underestimated the total mass of particulate matter present as compared to the centrifugation approach. This has serious implications forthe interpretation of results for "colloidal" and "soluble" fractions from soil suspensions and other environmental matrices obtained using the standard membrane approach. The results also show that sedimentation FFF has great potential as a robust and relatively mild technology for studying size distributions in the "colloidal" range for soil suspensions and other aquatic matrices.

Centrifugation↗