Immunomagnetic isolation of Xanthomonas campestris pv. pelargonii

Immunomagnetic fishing was developed as an improved procedure for increasing the bacterial target to non-target recovery ratio in suspensions containing mixtures of target and non-target organisms. A cell suspension containing the target Xanthomonas campestris pv. pelargonii and non-target organisms, is treated with rabbit polyclonal antiserum against X.c. pv. pelargonii and incubated for 1 h. The suspension is then mixed with paramagnetic iron oxide particles coated with goat anti-rabbit antibodies (immunomagnetic particles). After incubation, the polished surface of a 14 mm diameter neodymium supermagnet is placed at the air-water interace and the magnetic particles are attracted to the magnet. After all visible magnetic particles have attached to the bottom of the magnet, the magnet is dipped in sterile buffer to remove non-target organisms. The magnet with attached magnetic particles is rubbed evenly over an agar surface to dislodge the particles and attached bacteria. Conventional immunomagnetic isolation (immunomagnetic attraction) and immunomagnetic fishing were compared, for the recovery of the target organism in geranium leaf washings spiked with X.c. pv. pelargonii. With immunomagnetic attraction and immunomagnetic fishing, bacterial non-target organisms were reduced to 11.4 and 1.5% of the initial population, respectively, whereas the target was only reduced to 63.7 and 53.8%.     

Cell clarification and size separation using continuous countercurrent magnetophoresis

Nonmagnetic microparticles (e.g., cells, polymer beads) immersed in a magnetic fluid (ferrofluid) under a nonuniform magnetic field experience a magnetophoretic force in the direction of decreasing magnetic field strength. This phenomenon was exploited in the development of a continuous magnetophoretic countercurrent separation for the removal and concentration of micron-sized particles from aqueous suspensions, and in particular as a viable approach for cell clarification of raw fermentation broth. A magnetic fluid is added to the cell suspension, the mixture is introduced to the magnetic separator, which consists of an open flow tube passing between pairs of magnets that move in a direction counter to the flow of the suspension. The cells are pushed ahead of the magnet pairs owing to the magnetophoretic forces acting on them, collected in a tube upstream of the feed injection point, and removed as a concentrated suspension for further treatment.     

Magnetic field design for selecting and aligning immunomagnetic labeled cells

BACKGROUND: Recently we introduced the CellTracks cell analysis system, in which samples are prepared based on a combination of immunomagnetic selection, separation, and alignment of cells along ferromagnetic lines. Here we describe the underlying magnetic principles and considerations made in the magnetic field design to achieve the best possible cell selection and alignment of magnetically labeled cells. Materials and Methods Computer simulations, in combination with experimental data, were used to optimize the design of the magnets and Ni lines to obtain the optimal magnetic configuration. RESULTS: A homogeneous cell distribution on the upper surface of the sample chamber was obtained with a magnet where the pole faces were tilted towards each other. The spatial distribution of magnetically aligned objects in between the Ni lines was dependent on the ratio of the diameter of the aligned object and the line spacing, which was tested with magnetically and fluorescently labeled 6 microm polystyrene beads. The best result was obtained when the line spacing was equal to or smaller than the diameter of the aligned object. CONCLUSIONS: The magnetic gradient of the designed permanent magnet extracts magnetically labeled cells from any cell suspension to a desired plane, providing a homogeneous cell distribution. In addition, it magnetizes ferro-magnetic Ni lines in this plane whose additional local gradient adds to the gradient of the permanent magnet. The resultant gradient aligns the magnetically labeled cells first brought to this plane. This combination makes it possible, in a single step, to extract and align cells on a surface from any cell suspension.     

High gradient magnetic separation of erythrocytes

The high gradient magnetic separation technique has been applied to separate paramagnetic erythrocytes from a cell suspension that also contained diamagnetic cells. Paramagnetism was induced in the red blood cells by oxidizing the iron atoms in the cell hemoglobin to the ferric state (methemoglobin). Diamagnetic cells were either untreated erythrocytes, containing oxyferrohemoglobin, or leukocytes in a suspension of mouse spleen cells. Cell suspensions were passed through a column containing 40 micron diameter stainless steel wire in a high magnetic field (33 kG). The paramagnetic cells were retained on the surface of the wire while the diamagnetic cells passed through. Elution of the paramagnetic cells was accomplished by removing the column from the magnet, in effect turning off the field.     

Sorting of chromosomes by magnetic separation

Summary Chromosomes were isolated from Chinese hamster x human hybrid cell lines containing four and nine human chromosomes. Human genomic DNA was biotinylated by nick translation and used to label the human chromosomes by in situ hybridization in suspension. Streptavidin was covalently coupled to the surface of magnetic beads and these were incubated with the hybridized chromosomes. The human chromosomes were bound to the magnetic beads through the strong biotin-streptavidin complex and then rapidly separated from nonlabeled Chinese hamster chromosomes by a simple permanent magnet. The hybridization was visualized by additional binding of avidin-FITC (fluorescein) to the unoccupied biotinylated human DNA bound to the human chromosomes. After magnetic separation, up to 98% of the individual chromosomes attached to magnetic beads were classified as human chromosomes by fluorescence microscopy.     

Decreased chemotaxis of human peripheral phagocytes exposed to a strong static magnetic field

The chemotaxis of human peripheral phagocytes, neutrophils and monocytes was examined in a strong static magnetic field (0.317+/-0.012 Tesla). The chemotaxis of the suspension of purified neutrophils and monocytes was tested in the Boyden chamber using C5a as a chemotactic signal. The chambers were placed into a temperature regulated (36.6 degrees C) equipment producing a strong static magnetic field (0.317 Tesla) for 60 minutes. The movement of cells proceeded into a nitrocellulose membrane toward the north-pole of the magnet, i.e. in the direction of the Earth's gravitational pull. The C5a induced chemotaxis of human neutrophils decreased significantly in the strong static magnetic field. Monocytes were not significantly effected. The strong static magnetic field decreased the chemotactic movement of neutrophils and this phenomenon may have implications when humans are exposed to magnetic resonance imaging for extended periods of time.     

A Criterion for the Complete Deposition of Magnetic Beads on the Walls of Microchannels

This paper analyzes numerical simulations of the trajectories of magnetic beads in a microchannel, with a nearby permanent cubical magnet, under different flow and magnetic conditions. Analytically derived local fluid velocities and local magnetic forces have been used to track the particles. A centered position and a lateral position of the magnet above the microchannel are considered. The computed fractions of deposited particles on the walls are compared successfully with a new theoretically derived criterion that imposes a relation between the sizes of the magnet and the microchannel and the particle Stokes and Alfvén numbers to obtain the complete deposition of the flowing particles on the wall. In the cases in which all the particles, initially distributed uniformly across the section of the microchannel, are deposited on the walls, the simulations predict the accumulation of the major part of particles on the wall closest to the magnet and near the first half of the streamwise length of the magnet.     

Magnetostatic Field System for Uniform Cell Cultures Exposure

The aim of the present work has been the design and the realization of a Magnetostatic Field System for Exposure of Cell cultures (MaFiSEC) for the uniform and the reproducible exposure of cell cultures to static magnetic fields (SMFs) of moderate magnetic induction. Experimental and computer-simulated physical measurements show that MaFiSEC: i) generates a SMF with magnetic induction that can be chosen in the range of 3 to 20 mT; ii) allows the uniform SMF exposure of cells growing in adhesion and in suspension; iii) is cheap and easy to use. The efficacy and reproducibility of MaFiSEC has been tested by comparing the biological effects exerted on isolated human lymphocytes by 72 h of exposure to a magnet (i.e. Neodymium Magnetic Disk, NMD) placed under the culture Petri dish. Lymphocytes morphology, viability, cell death, oxidative stress and lysosomes activity were the parameters chosen to evaluate the SMF biological effects. The continuous exposure of cells to a uniform SMF, achieved with MaFiSEC, allows highly reproducible biochemical and morphological data.     

适用于磁压榨吻合的平移钳的设计

目的:提出了磁压榨吻合操作中可用于磁体夹持固定的平移钳的结构设计方案。方法:分析本团队多种原创性磁压榨吻合技术操作中存在的缺陷和不足,认为目前磁压榨吻合操作中依靠术者徒手控制或利用现有血管钳对磁体进行夹持和固定,是造成操作中诸多不便的主要原因。从力学角度分析了常规血管钳夹持磁体易导致磁体滑脱的原因,指出设计加工磁吻合专用钳是解决目前磁吻合操作不便的有效措施。在此基础上提出了利用平移钳来固定磁体的结构设计方案,并从力学角度进行了受力分析,设计出了用于磁压榨吻合操作中磁体夹持固定的平移钳。结果:平移钳的结构设计能够稳定夹持磁体,避免磁体滑脱;同时,借助齿轮传动结构控制钳头的平行移动能够更好地控制磁体的精准对吸和分离,可进一步简化操作,避免副损伤,节省手术操作时间。结论:平移钳能满足多种形状及大小的磁体的夹持和固定,可有效控制磁体的吸合与分离,极大地方便操作。该平移钳加工简单,使用方便,有助于推动磁压榨吻合技术在临床广泛开展。     

Optimization of static magnetic field parameters improves analgesic effect in mice

The present study deals with the analgesic effect induced by static magnetic fields (SMF) in mice exposed to the field with their whole body. It discusses how the effect depends on the distribution of the magnetic field, that is, on the specification and arrangement of the applied individual permanent magnets. A critical analysis of different magnet arrangements is given. As a result the authors propose a magnet arrangement recipe that achieves an analgesic effect of over 80% in the writhing test. This is a widely accepted screening method for animal pain and predictor of human experimental results. As a non-drug, non-invasive, non-contact, non-pain, non-addictive method for analgesia with immediate and long-lasting effect based on the stimulus of the endogenous opioid network, the SMF treatment may attract the attention of medical doctors, nurses, magnet therapists, veterinarians, physiotherapists, masseurs, and fitness trainers among others.     

Quantitative Modeling and Optimization of Magnetic Tweezers

Magnetic tweezers are a powerful tool to manipulate single DNA or RNA molecules and to study nucleic acid-protein interactions in real time. Here, we have modeled the magnetic fields of permanent magnets in magnetic tweezers and computed the forces exerted on superparamagnetic beads from first principles. For simple, symmetric geometries the magnetic fields can be calculated semianalytically using the Biot-Savart law. For complicated geometries and in the presence of an iron yoke, we employ a finite-element three-dimensional PDE solver to numerically solve the magnetostatic problem. The theoretical predictions are in quantitative agreement with direct Hall-probe measurements of the magnetic field and with measurements of the force exerted on DNA-tethered beads. Using these predictive theories, we systematically explore the effects of magnet alignment, magnet spacing, magnet size, and of adding an iron yoke to the magnets on the forces that can be exerted on tethered particles. We find that the optimal configuration for maximal stretching forces is a vertically aligned pair of magnets, with a minimal gap between the magnets and minimal flow cell thickness. Following these principles, we present a configuration that allows one to apply ≥40 pN stretching forces on ≈1-μm tethered beads.     

High-intensity static magnetic fields modulate skin microcirculation and temperature in vivo

We investigated the acute effect of static magnetic fields of up to 8 T on skin blood flow and body temperature in anesthetized rats. These variables were measured prior to, during, and following exposure to a magnetic field in a superconducting magnet with a horizontal bore. The dorsal skin was transversely incised for 1 cm to make a subcutaneous pocket. Probes of a laser Doppler flowmeter and a thermistor were inserted into the pocket and positioned at mid-dorsum to measure skin blood flow and temperature. Another thermistor probe was put into the rectum to monitor rectal temperature. After baseline measurement outside the magnet, the rat was inserted into the bore for 20 min so that mid-dorsum was exactly positioned at the center, where the magnetic field was nearly homogeneous. Post-exposure changes were then recorded for 20 min outside the bore. Sham-exposed animals were submitted to exactly the same conditions, except that the superconducting magnet was not energized. Skin blood flow and temperature decreased significantly during magnetic field exposure and recovered after removal of the animal from the magnet. The rectal temperature showed a tendency to decrease while the animal was in the magnet. The microcirculatory and thermal reactions in the present study were consistent and agreed with some of the predictions based on mathematical simulations and model experiments.     

Molecular nanomagnetism in Florence: Advancements and perspectives

Magnetic materials exhibiting magnetic hysteresis in the absence of magnetic order are fascinating systems for fundamental science and possible applications but the temperature at which the magnetic memory is observed remains rather low. The intrinsic limits of the single molecule magnet and single chain magnet approaches are here briefly discussed as well as possible perspectives in molecular nanomagnetism.     

Tapping culture--an improved method for cell suspension culture.

A new culture vessel was designed for cell suspension culture. A silicone-covered magnet bar fixed by one end to the side wall of the bottle was held horizontally a short distance from the bottom. A standard type magnetic stirrer was used. In contrast to the conventional horizontal movement of "stirring" in cultures the bar moves vertically with a "tapping" motion. This improvement resulted in less cell injury, higher rate of cell proliferation and formation of fewer bubbles than in the conventional type. Nine cell types were simultaneously cultivated in tapping, stirring and stationary culture. All cell types proliferated more luxuriously in tapping cultures then in stirring cultures. Serial cultivation of cells in tapping cultures was also successful.     

A novel technique for in situ aggregation of Gluconobacter oxydans using bio‐adhesive magnetic nanoparticles

Here, we present a novel technique to immobilize magnetic particles onto whole Gluconobacter oxydans in situ via a synthetic adhesive biomimetic material inspired by the protein glues of marine mussels. Our approach involves simple coating of a cell adherent polydopamine film onto magnetic nanoparticles, followed by conjugation of the polydopamine‐coated nanoparticles to G. oxydans which resulted in cell aggregation. After optimization, 21.3 mg (wet cell weight) G. oxydans per milligram of nanoparticle was aggregated and separated with a magnet. Importantly, the G. oxydan aggregates showed high specific activity and good reusability. The facile approach offers the potential advantages of low cost, easy cell separation, low diffusion resistance, and high efficiency. Furthermore, the approach is a convenient platform technique for magnetization of cells in situ by direct mixing of nanoparticles with a cell suspension. Biotechnol. Bioeng. 2012; 109: 2970–2977. © 2012 Wiley Periodicals, Inc.     

Effects of magnetic fields on fyke net performance

The effects of magnetic fields generated by ferritic magnets and mounted in entrances to fyke nets were studied in 1999–2002. With respect to all fish species trapped (perch, pike, roach, rudd, bleak, bream, ruffe) except eel, the catch potential of magnet‐rigged fyke nets was found to be significantly higher (almost 50% on average) when compared with controls. Differences in magnet placement (N or S poles inside the traps) proved to have no significant importance on the number of fish that chose to enter the fyke nets. Specific behavioural mechanisms in adult fish towards magnetic fields as well as application of fishing gear magnet rigging in fisheries practice in inland waters are discussed.     

基于磁驱动技术的空肠营养管的设计

目的:为解决目前床旁鼻肠管快速置入成功率低这一临床难题,该文提出一种基于磁驱动技术的空肠营养管的设计。方法:分析了现有空肠营养管置管过程中的动力因素,结合磁驱动技术原理,提出了通过体外旋转磁场带动体内感应磁体螺旋式前进的设计方案,以期缩短空肠营养管的飘管时间。结果:该设计包括磁性空肠营养管和体外磁力驱动装置两部分。其中磁性空肠营养管由管体和感应磁头两部分构成,感应磁头包括磁体内核和硅胶外壳。磁力驱动装置由多极磁体和手持式微型电机组成。操作时通过体外磁力驱动装置发出大旋转磁场带动空肠营养管的感应磁头做轴向旋转运动,可加速空肠营养管在肠道内的移动,达到缩短飘管时间的目的。结论:该设计基于磁驱动技术原理,设计巧妙,符合磁力学原理,操作简单,具有临床应用潜力。     

High gradient magnetic separation of rosette-forming cells

High Gradient Magnetic Separation (HGMS) is a rapid and straightforward technique that has previously been proven effective in extracting erythrocytes from a flowing cell suspension if the red cell hemoglobin is in a paramagnetic state. In this work it was applied to the enrichment of the small population (<2%) of splenocytes from an immune mouse that bound sheep red cells to form rosettes. Samples flowed through the HGMS column in a strong magnetic field where rosettes and free sheep cells were selectively retained. These were subsequently eluted by simply removing the magnetic field. The process required 20–30 min per mouse spleen. Rosettes in the initial sample and in the fractions that passed through, or were retained by, the column were enumerated under the microscope. Under the conditions used here, the retained and eluted cells typically showed a 20–50-fold increase in the frequency of rosetted cells, and the cells that passed through the magnet showed 90–100% depletion of rosettes. The recovery of intact rosettes and the overall cell recovery were generally both in the range of 80–90%.     

Induction of primary root curvature in radish seedlings in a static magnetic field

Primary roots of radish (Raphanus sativus L.) seedlings were exposed to an inhomogeneous static magnetic field generated by a permanent magnet, during continuous rotation on a 0.06 rpm clinostat, thereby reducing the unilateral influence of gravity. The roots responded tropically to the static magnetic field with the tropism appearing to be negative. These roots responded significantly (P < 0.05) to the south pole of the magnet. The significant tropic response was found for a magnetic flux density of 13-68 mT, for a field gradient of 1.8-14.7 T/m, and for the product of magnetic field and field gradient of 0.023-1.0 T(2)/m. A small, but insignificant, response of the roots to the north pole has also been found.     

Effects of non-uniform static magnetic fields on the rate of myosin phosphorylation

The effect on myosin phosphorylation from exposure to a magnetic field generated by an array of four permanent magnets was investigated. Two lateral positions in the non-uniform field over the array were explored, each at four vertical distances over the surface of the device. The rate of myosin phosphorylation was found to depend on the position laterally over the array as well as the distance from the device surface. The square magnet array was comprised of axially magnetized, cylindrical NdFeB permanent magnets arranged with poles of alternating polarity in a plane (MagnaBloc trade mark therapeutic device). Detailed dosimetry of the magnet array was compiled: the magnetic flux density averaged over the exposure volume spanned the range 0.7-86 mT for the eight different exposure positions. The corresponding range for the absolute field gradient was 0.4-20 T/m. Comparing the dosimetry to the experimental outcome, our results imply that magnetic field amplitude alone is not sufficient to describe the influence of the field in this preparation.