Magnetic cell separation: characterization of magnetophoretic mobility

Magnetic cell separation has become a popular technique to enrich or deplete cells of interest from a heterogeneous cell population. One important aspect of magnetic cell separation is the degree to which a cell binds paramagnetic material. It is this paramagnetic material that imparts a positive magnetophoretic mobility to the target cell, thus allowing effective cell separation. A mathematical relationship has been developed to correlate magnetic labeling to the magnetophoretic mobility of an immunomagnetically labeled cell. Four parameters have been identified that significantly affect magnetophoretic mobility of an immunomagnetically labeled cell: the antibody binding capacity (ABC) of a cell population, the secondary antibody amplification (psi), the particle-magnetic field interaction parameter (DeltachiV(m)), and the cell diameter (D(c)). The ranges of these parameters are calculated and presented along with how the parameters affect the minimum and maximum range of magnetophoretic mobility. A detailed understanding of these parameters allows predictions of cellular magnetophoretic mobilities and provides control of cell mobility through selection of antibodies and magnetic particle conjugates.     

Control of magnetophoretic mobility by susceptibility-modified solutions as evaluated by cell tracking velocimetry and continuous magnetic sorting

With the analytical expression for the magnetophoretic mobility of an ideal, linearly polarizable sphere undergoing creeping motion in viscous medium, we have shown that both attractive and repulsive motions are possible in the magnetic field. We have validated theoretical predictions using magnetic monodisperse microspheres of 5.2-microm diameter and nonmagnetic polystyrene microspheres of 6.99-microm diameter suspended in solutions of paramagnetic ions. The microsphere magnetophoretic mobility was measured using a modified particle tracking velocimetry system, developed in-house and called a cell tracking velocimeter. The product of measured mobility and viscosity agrees well with the theoretical prediction, differing only by approximately 11%. Further, a 26% increase in resolution between magnetic and nonmagnetic particle distributions was evaluated when paramagnetic ion carrier was used instead of water. Continuous particle sorting based on differences in magnetophoretic mobility was performed with another device developed by us, the quadrupole magnetic flow sorter (QMS). In the QMS, the introduction of paramagnetic ions into the carrier was effective in suppressing nonspecific crossover (i.e., the transport of low-mobility particles into the magnetic particle fraction) in particles and in biologically relevant red blood cells and thus showed promise as a means of increasing the purity of the magnetic separation.     

Multifunctional Silver‐Embedded Magnetic Nanoparticles as SERS Nanoprobes and Their Applications

In this study, surface‐enhanced Raman spectroscopy (SERS)‐encoded magnetic nanoparticles (NPs) are prepared and utilized as a multifunctional tagging material for cancer‐cell targeting and separation. First, silver‐embedded magnetic NPs are prepared, composed of an 18‐nm magnetic core and a 16‐nm‐thick silica shell with silver NPs formed on the surface. After simple aromatic compounds are adsorbed on the silver‐embedded magnetic NPs, they are coated with silica to provide them with chemical and physical stability. The resulting silica‐encapsulated magnetic NPs (M‐SERS dots) produce strong SERS signals and have magnetic properties. In a model application as a tagging material, the M‐SERS dots are successfully utilized for targeting breast‐cancer cells (SKBR3) and floating leukemia cells (SP2/O). The targeted cancer cells can be easily separated from the untargeted cells using an external magnetic field. The separated targeted cancer cells exhibit a Raman signal originating from the M‐SERS dots. This system proves to be an efficient tool for separating targeted cells. Additionally, the magnetic‐field‐induced hot spots, which can provide a 1000‐times‐stronger SERS intensity due to aggregation of the NPs, are studied.     

Dynamics of magnetophoretic separation of a suspension of slightly magnetic microparticles in a high-gradient field of a magnetized rod

A macroscopic model of the process of magnetophoretic separation of slightly magnetic particles from a suspension under the action of a high-gradient magnetic field with account for the particle size is proposed. The mechanisms of separation of dia- and paramagnetic particles from a narrow fluid layer in an inhomogeneous field of a magnetized rod have been investigated. Translated from Inzhenerno-Fizicheskii Zhurnal, Vol. 82, No. 2, pp. 221–226, March–April, 2009.     

Magnetophoretic velocimetry of manganese(II) in a single microdroplet in a flow system under a high gradient magnetic field generated with a superconducting magnet

An experimental system for magnetophoretic velocimetry, which could determine the volume magnetic susceptibility of a single particle dispersed in a liquid phase from a magnetophoretic velocity, has been developed. A micrometer-sized high-gradient magnetic field could be generated in a capillary by a pair of iron pole pieces in a superconducting magnet (10 T). The magnetophoretic behavior of a single particle in a capillary flow system was investigated under the inhomogeneous magnetic field. From the magnetophoretic velocity of a polystyrene latex particle dispersed in a MnCl2 aqueous solution, the product of the magnetic flux density and the gradient, B(dB/dx), was determined as a function of the position along the capillary. The maximum value of B(dB/dx) was 4.7 x 10(4) T2 m(-1), which was approximately 100 times higher than that obtained by two Nd-Fe-B permanent magnets (0.4 T). Organic droplets extracting manganese(II) with 2-thenoyltrifluoroacetone and tri-n-octylphosphine oxide from MnCl2 solution were used as test samples. The difference of the volume magnetic susceptibility between the droplet and the medium could be determined from the magnetophoretic velocity. This method allowed us to continuously measure a volume magnetic susceptibility of 10-6 level for a picoliter droplet and to determine manganese(II) in the single droplet at the attomole level.     

On-chip free-flow magnetophoresis: continuous flow separation of magnetic particles and agglomerates

The separation of magnetic microparticles was achieved by on-chip free-flow magnetophoresis. In continuous flow, magnetic particles were deflected from the direction of laminar flow by a perpendicular magnetic field depending on their magnetic susceptibility and size and on the flow rate. Magnetic particles could thus be separated from each other and from nonmagnetic materials. Magnetic and nonmagnetic particles were introduced into a microfluidic separation chamber, and their deflection was studied under the microscope. The magnetic particles were 2.0 and 4.5 microm in diameter with magnetic susceptibilities of 1.12 x 10(-4) and 1.6 x 10(-4) m(3) kg(-1), respectively. The 4.5-microm particles with the larger susceptibility were deflected further from the direction of laminar flow than the 2.0-microm magnetic particles. Nonmagnetic 6-microm polystyrene beads, however, were not deflected at all. Furthermore, agglomerates of magnetic particles were found to be deflected to a larger extent than single magnetic particles. The applied flow rate and the strength and gradient of the applied magnetic field were the key parameters in controlling the deflection. This separation method has a wide applicability since magnetic particles are commonly used in bioanalysis as a solid support material for antigens, antibodies, DNA, and even cells. Free-flow magnetophoretic separations could be hyphenated with other microfluidic devices for reaction and analysis steps to form a micro total analysis system.     

Reduction of hexavalent chromium by Pannonibacter phragmitetus LSSE-09 coated with polyethylenimine-functionalized magnetic nanoparticles under alkaline conditions

A novel cell separation and immobilization method for Cr (VI)-reduction under alkaline conditions was developed by using superparamagnetic Fe(3)O(4) nanoparticles (NPs). The Fe(3)O(4) NPs were synthesized by coprecipitation followed by modification with sodium citrate and polyethyleneimine (PEI). The surface-modified NPs were monodispersed and the particle size was about 15 nm with a saturation magnetization of 62.3 emu/g and an isoelectric point (pI) of 11.5 at room temperature. PEI-modified Fe(3)O(4) NPs possess positive zeta potential at pH below 11.5, presumable because of the high density of amine groups in the long chains of PEI molecules on the surface. At initial pH 9.0, Pannonibacter phragmitetus LSSE-09 cells were immobilized by PEI-modified NPs via electrostatic attraction and then separated with an external magnetic field. Compared to free cells, the coated cells not only had the same Cr (VI)-reduction activity but could also be easily separated from reaction mixtures by magnetic force. In addition, the magnetically immobilized cells retained high specific Cr (VI)-reduction activity over six batch cycles. The results suggest that the magnetic cell separation technology has potential application for Cr (VI) detoxification in alkaline wastewater.     

Magnetophoretic Potential of a Plane‐Ordered System of Ferrocylinders. 2. Rectangular Cylinders

The magnetophoretic properties of a system of equidistant identical rectangular ferromagnetic cylinders located in one plane and exposed to a uniform magnetic field are considered. The analytical solution of the problem on distribution of the magnetic field and the magnetophoretic potential in magnetization of the system to saturation along the packing plane and across it is obtained. The influence of geometric and magnetic parameters on the magnetophoretic properties of the system is investigated.     

Magnetophoretic velocimetry of manganese(II) in a single emulsion droplet at the femtomole level.

We developed a new experimental technique named magnetophoretic velocimetry to determine a small amount of paramagnetic species in a single microdroplet. The magnetophoretic velocity of an aqueous droplet containing paramagnetic metal ion dispersed in an organic medium could response to a very small amount of the metal ion under an inhomogeneous magnetic field. The paramagnetic droplet (2 approximately 8 microm diam) used as a test sample in this study was the aqueous droplet of manganese(II) chloride dispersed in ethylbenzoate whose density was nearly equal to water. A pair of small Nd-Fe-B magnets placed with a gap of 400 microm generated an inhomogeneous magnetic field between the edges, at which the product of the magnetic flux density and the gradient, B(dB/dx), was as high as 410 T2 m(-1). When a silica capillary containing the emulsion was inserted into the gap between the magnets, the magnetophoretic migration of the droplets was observed with a video microscope. The magnetophoretic velocity divided by the squared radius of the droplet was proportional to the MnCl2 concentration in the droplet, as predicted by a theoretical calculation. The estimated detection limit in this simple method was lower than 10(-16) mol for manganese(II).     

Paramagnetic capture mode magnetophoretic microseparator for blood cells

The paper presents the characterisation of a continuous paramagnetic capture (PMC) mode magnetophoretic microseparator for separating red and white blood cells from whole blood based on their native magnetic properties. The PMC microseparator separates the blood cells using a high-gradient magnetic separation method without the use of additives such as magnetic tagging. The microseparator is fabricated using microfabrication technology, enabling the integration of micro-scale magnetic flux concentrators in an aqueous micro-environment. Experimental results show that the PMC microseparator can continuously separate out 91.1% of red blood cells from whole blood within 5 min, using an external magnetic flux of 0.2 T from a permanent magnet. Monitoring of white blood cells dyed with a fluorescent probe shows that 87.7% of white blood cells are separated out by the 0.2 T external magnetic flux applied to the PMC microseparator.     

Magnetophoretic immunoassay of allergen-specific IgE in an enhanced magnetic field gradient

We demonstrate a novel magnetophoretic immunoassay of allergen-specific immunoglobulin E (IgE) based on the magnetophoretic deflection velocity of a microbead that is proportional to the associated magnetic nanoparticles under enhanced magnetic field gradient in a microchannel. In this detection scheme, two types of house dust mites, Dermatophagoides farinae (D. farinae) and Dermatophagoides pteronyssinus (D. pteronyssinus), were used as the model allergens. Polystyrene microbeads were conjugated with each of the mite extracts followed by incubation with serum samples. The resulting mixture was then reacted with magnetic nanoparticle-conjugated anti-human IgE for detection of allergen-specific IgE by using sandwich immuno-reactions. A ferromagnetic microstructure combined with a permanent magnet was employed to increase the magnetic field gradient ( approximately 10(4) T/m) in a microfluidic device. The magnetophoretic velocities of microbeads were measured in a microchannel under applied magnetic field, and the averaged velocity was well correlated with the concentration of allergen-specific IgE in serum. From the analysis of pooled sera obtained from 44 patients, the detection limits of the allergen-specific human IgEs for D. farinae and D. pteronyssinus were determined to be 565 (0.045 IU/mL) and 268 fM (0.021 IU/mL), respectively. These values are 1 order of magnitude lower than those by a conventional CAP system. For evaluation of reproducibility and accuracy, unknown sera were subjected to a blind test by using the developed assay system, and they were compared with the CAP system. As a result, coefficient of variance was less than 10%, and the developed method enabled a fast assay with a tiny amount of serum ( approximately 10 microL).     

Setting up High Gradient Magnetic Separation for combating eutrophication of inland waters

To find new approaches to devise technologies for handling with eutrophication of inland waters is a global challenge. Separation of the P from water under conditions of continuous flow is proposed as an alternative and effective method. This work is based on using highly magnetic particles as the seeding adsorbent material and their later removal from solution by High Gradient Magnetic Separation (HGMS). Contrast to other methods based on batch conditions, large volumes of water can be easily handled by HGMS because of decreasing retention times. This study identifies the best working conditions for removing P from solution by investigating the effects of a set of four different experimental variables: sonication time, flow rate (as it determines the retention time of particles in the magnetic field), magnetic field strength and the iron (Fe) particles/P concentration ratio. Additionally, the change of P removal efficiency with time (build up effect) and the possibility of reusing magnetic particles were also studied. Our results evidenced that while flow rate does not significantly affect P removal efficiency in the range 0.08-0.36 mL s(-1), sonication time, magnetic field strength and the Fe particles/P concentration ratio are the main factors controlling magnetic separation process.     

Gold nanoparticle enhanced electrochemiluminescence of CdS thin films for ultrasensitive thrombin detection

Interactions between surface plasmons (SP) of metallic surfaces and photoluminescence (PL) of semiconductor nanocrystal (S-NC) surfaces have been extensively investigated, and SP-induced PL enhancement has been used as a sensitive analytical technique. However, this SP induced electrochemiluminescence (ECL) enhancement is rarely studied. In this work, we report greatly enhanced ECL of CdS thin films by gold nanoparticles (Au NPs) for ultrasensitive detection of thrombin. The system was composed of a CdS NC film on glassy carbon electrode (GCE) as ECL emitter attached an aptamer of thrombin. Then, ssDNA-AuNP conjugates hybridized with the aptamer to form a separation length of ca. 12 nm between CdS NCs and Au NPs. The system showed 5-fold enhancement of ECL intensity as compared to that without Au NPs, which might be attributed to the long-distance interaction between the S-NCs and SPR field of noble metal nanoparticles (MNPs).We also found that the enhanced ECL could be influenced by the involving factors such as the separation distance, spectral overlap, and magnetic field. Such enhancement in combination with smart recognition of aptamer and target protein allowed us to construct an ultrasensitive aptasensor for attomolar detection of thrombin. The presence of target protein was reflected by the ECL signal decrease caused by the target-induced removal of ssDNA-AuNP conjugates. The decrease of ECL signal was logarithmically linear with the concentration of thrombin in a wide range from 100 aM to 100 fM. The principle described in this work could be also applied to many other bioassays.     

Magnetophoretic velocity modulation mass analysis of a single microparticle in an atmosphere

A new principle of the magnetophoretic velocity modulation mass analysis of microparticles, which can determine simultaneously the mass and magnetic susceptibility of a single microparticle, has been proposed, and the measurement system was constructed by applying a magnetophoretic force on a falling microparticle through a magnetic field gradient in an atmosphere. A polystyrene microparticle as a test particle adsorbed on a glass plate was selectively knocked off by a pulsed Nd:YAG laser impact into a narrow gap of pole pieces of permanent magnets having a magnetic field gradient with a maximum intensity of 850 T2 m(-1). The falling particle was irradiated by a He-Ne laser, and the scattered light was detected through a slit array mask as a function of time. A bundle of spiked signals of scattered light intensity was analyzed to obtain velocities, which gave acceleration and deceleration of the falling particle. On the basis of the equation of motion under the magnetic field gradient, the mass and magnetic susceptibility of the test particle were reasonably determined.     

磁性石墨烯的制备初探

目的制备磁性石墨烯,实现其定向排列,为进一步制备性能良好的热界面材料提供定向导热相。方法采用聚合物包覆和层层组装技术,在石墨烯表面均匀包覆带负电荷的聚磺苯乙烯(PSS)聚电解质层,然后在静电力作用下包覆一层带正电荷的聚二烯丙基二甲基氯化铵(PDDA)聚电解质。最后,借助静电力使带负电荷的磁性Fe3O4纳米粒子在附着有PDDA层的石墨烯表面形成均匀的致密覆盖层,得到磁性石墨烯,并在外磁场作用下使磁性石墨烯进行定向排列。结果采用聚电解质层层组装技术可有效地对石墨烯进行磁性化处理;粉状石墨烯比片状石墨烯定向排列效果明显;以片状石墨烯为原材料的实验条件下,加入聚电解PSS、PDDA的量越多,磁性化效果更佳。结论通过聚电解质层层组装技术可有效地对石墨烯进行磁性化处理,并在磁场作用下实现定向排列。     

Removal and reuse of Ag nanoparticles by magnetic polyaniline/Fe<Subscript>3</Subscript>O<Subscript>4</Subscript> nanofibers

For the treatment of wastewater containing Ag nanoparticles (NPs), PANI/Fe₃O₄ nanofibers were firstly prepared by a novel self-assemble. And then, the efficiency for the removal of Ag NPs from wastewater was investigated. The magnetic performance of PANI/Fe₃O₄ nanofibers could be optimized by adjusting the pH of the self-assemblied system. Under pH of 3, the as-prepared nanofibers exhibited the highest magnetism and also displayed good efficiency (>?12 mg g^?1) for the removal of Ag NPs. Importantly, the resulted product (PANI/Fe₃O₄/Ag composite) could act as a catalysis for cleaning durable pollutant, 4-nitrophenol. After 10 cycles, only slight decrease in rate constant was found, indicating excellent reusability. Those approaches provide a new way to merge the recovery of Ag NPs as pollutants and reuse of recovered Ag NPs as recyclable material for environmental remediation.     

A biotechnological perspective on the application of iron oxide nanoparticles

In recent decades,magnetic iron nanoparticles (NPs) have attracted much attention due to properties such as superparamagnetism,high surface area,large surface-to-volume ratio,and easy separation under external magnetic fields.Therefore,magnetic iron oxides have potential for use in numerous applications,including magnetic resonance imaging contrast enhancement,tissue repair,immunoassay,detoxification of biological fluids,drug delivery,hyperthermia,and cell separation.This review provides an updated and integrated focus on the fabrication and characterization of suitable magnetic iron NPs for biotechnological applications.The possible perspective and some challenges in the further development of these NPs are also discussed.     

Autonomous Magnetic Microrobots by Navigating Gates for Multiple Biomolecules Delivery

The precise delivery of biofunctionalized matters is of great interest from the fundamental and applied viewpoints. In spite of significant progress achieved during the last decade, a parallel and automated isolation and manipulation of rare analyte, and their simultaneous on‐chip separation and trapping, still remain challenging. Here, a universal micromagnet junction for self‐navigating gates of microrobotic particles to deliver the biomolecules to specific sites using a remote magnetic field is described. In the proposed concept, the nonmagnetic gap between the lithographically defined donor and acceptor micromagnets creates a crucial energy barrier to restrict particle gating. It is shown that by carefully designing the geometry of the junctions, it becomes possible to deliver multiple protein‐functionalized carriers in high resolution, as well as MCF‐7 and THP‐1 cells from the mixture, with high fidelity and trap them in individual apartments. Integration of such junctions with magnetophoretic circuitry elements could lead to novel platforms without retrieving for the synchronous digital manipulation of particles/biomolecules in microfluidic multiplex arrays for next‐generation biochips.     

Optical thin film filters of colloidal gold and silica nanoparticles prepared by a layer-by-layer self-assembly method

A novel fabrication method for optical thin film filters based on the self-organization of alternating layers of colloidal gold and silica nanoparticles (NP) is reported. The filter is designed to work in the deep-UV to visible spectral range. The spectral absorption peaks are tuned by three parameters: the organic capping ligand of the gold NPs (citrate, chitosan, poly (diallyl-dimethylammonium)-chloride or PDDA); the capping environment (bare, chitosan, or PDDA) of the silica NPs and the thickness of the film. Precise control of the transmission color (less than 1% color distance per layer), is achieved by changing the film thickness. Exploitation of the self-assembly process should lead to the facile production of highly reliable large area thin film optical filters at considerably lower costs.     

Influence of carboxyl group formation on ammonia adsorption of NiO-templated nanoporous carbon surfaces

The scope of this work was to control the surface functional groups of nanoporous carbons (NPs) by oxidizing agents (nitric acid and hydrogen peroxide) treatments and to investigate the relation between carboxyl group and ammonia removal efficiency. The NPs were directly prepared from a cation exchange resin by the carbonization of a mixture with Ni acetate at 900 °C. N₂/-196 °C adsorption, Boehm's titrations, and X-ray photoelectron spectroscopy (XPS) analyzes were employed to confirm the physicochemical properties of NPs. The ammonia removal efficiency was confirmed by temperature programmed desorption (TPD) technique. In the result, the oxygen content of NPs increased after various treatments and the highest content of carboxyl group formation appeared at a 2:3 volume ratio of HNO₃/H₂O₂. It was also found that the oxidation treatment led to an increase in ammonia removal efficiency of NPs, mainly due to an increase of acid oxygen functional groups (such as carboxyl) on NPs surfaces.