Magnetophoresis and magnetic susceptibility of HeLa tumor cells

A method for measuring the magnetic susceptibility of separate cells has been theoretically developed and implemented in experiments. It is based on fitting the videocomputer and predicted data for the integral motion curve of cells that settle in a fluid along a thin ferromagnetic bar by the action of a uniform magnetic field. The magnetic susceptibility of HeLa tumor cells and culture medium 199 has been measured.     

Cell volume dependence of 1H spin-echo NMR signals in human erythrocyte suspensions. The influence of in situ field gradients

The 1H spin-echo NMR signal amplitudes and intensities of low molecular weight solutes in the cytoplasm and extracellular fluid of suspensions of human erythrocytes were shown to depend on the osmotic pressure of the media. At low osmotic pressure (220 mosM/kg) freeze-thaw lysis of the cells resulted in signal enhancement which was greatest for extracellular molecules, but both intra- and extracellular species were almost equally enhanced at 580 mosM/kg. This effect is due to field gradients formed at cell boundaries as a result of differences in magnetic susceptibility between the medium and the cytoplasm. T2 values measured using the Carr-Purcell-Meiboom-Gill pulse sequence, with tau = 0.0003 s, depended little on cell volume and absolute changes in volume magnetic susceptibility were also small. The mean field gradients, calculated from data obtained on cell suspensions at different osmotic pressures, were in the range 0.25-1.98 G/cm and 0.89-2.09 G/cm for intra- and extracellular compartments, respectively. The maintenance of isotonicity of the extracellular fluid during metabolic studies of cell suspensions is important in order to avoid artefacts in the determination of metabolite concentrations when using the spin-echo technique. Conversely it may be possible to perform transport measurements using spin-echo NMR to monitor the cell volume changes which occur during the transmembrane migration of molecules.     

Quantitative characterization of the regulation of iron metabolism in glioblastoma stem-like cells using magnetophoresis

This study focuses on different iron regulation mechanisms of glioblastoma (GBM) cancer stem-like cells (CSCs) and non-stem tumor cells (NSTCs) using multiple approaches: cell viability, density, and magnetophoresis. GBM CSCs and NSTCs were exposed to elevated iron concentration, and their magnetic susceptibility was measured using single cell magnetophoresis (SCM), which tracks the magnetic and settling velocities of thousands of individual cells passing through the magnetic field with a constant energy gradient. Our results consistently demonstrate that GBM NSTCs have higher magnetic susceptibility distribution at increased iron concentration compared with CSCs, and we speculate that it is because CSCs have the ability to store a high amount of iron in ferritin, whereas the free iron ions inside the NSTCs lead to higher magnetic susceptibility and reduced cell viability and growth. Further, their difference in magnetic susceptibility has led us to pursue a separate experiment using a quadrupole magnetic separator (QMS), a novel microfluidic device that uses a concentric channel and permanent magnets in a special configuration to separate samples based on their magnetic susceptibilities. GBM CSCs and NSTCs were exposed to elevated iron concentration, stained with two different trackers, mixed and introduced into QMS; subsequently, the separated fractions were analyzed by fluorescent microscopy. The separation results portray a successful label-less magnetic separation of the two populations.     

Effect of temperature and pH on magnetic properties of human erythrocytes

Erythrocytes magnetic susceptibility depending on temperature and pH value of buffer were studied by analysing individual erythrocytes moving in a medium in the nonhomogeneous magnetic field near a transversely magnetized thin wire.     

Magnetic susceptibility of microorganisms.

Total magnetic susceptibility of 13 species and varieties of bacteria was investigated using the relative method of Guy. It has been established that the index of magnetic susceptibility is a constant characteristic of bacteria. Total magnetic susceptibility ranged from --0.3295.10(-6) in Escherichia P678 to --0.4965.10(-6) in Proteus. It has also been established that magnetic susceptibility changes during long-term passages of bacteria in fluctuating +/- 0.1 Oe) magnetic field. This is suggestive of a low threshold of their magnetic susceptibility and permits a rough assessment of the importance of fluctuations of the geomagnetic field for the viability of microbes.     

Application of spin-echo nuclear magnetic resonance to whole-cell systems. Membrane transport.

A new method for studying membrane transport is presented. High resolution n.m.r. is used to measure the distribution of small molecules between the intracellular and extracellular compartments. The method uses spin-echo techniques and relies on a difference in the magnetic susceptibility of the media inside and outside of cells. It also provides simultaneous information on the metabolic status of the cell. The method is illustrated by a study of alanine and lactate transport in the human erythrocyte.     

Cell volume dependence of 1H spin-echo NMR signals in human erythrocyte suspensions: The influence of in situ field gradients

The ¹H spin-echo NMR signal amplitudes and intensities of low molecular weight solutes in the cytoplasm and extracellular fluid of suspensions of human erythrocytes were shown to depend on the osmotic pressure of the media. At low osmotic pressure (220 mosM/kg) freeze-thaw lysis of the cells resulted in signal enhancement which was greatest for extracellular molecules, but both intra- and extracellular species were almost equally enhanced at 580 mosM/kg. This effect is due to field gradients formed at cell boundaries as a result of differences in magnetic susceptibility between the medium and the cytoplasm. T₂ values measured using the Carr-Purcell-Meiboom-Gill pulse sequence, with τ = 0.0003 s, depended little on cell volume and absolute changes in volume magnetic susceptibility were also small. The mean field gradients, calculated from data obtained on cell suspensions at different osmotic pressures, were in the range 0.25–1.98 G/cm and 0.89–2.09 G/cm for intra- and extracellular compartments, respectively. The maintenance of isotonicity of the extracellular fluid during metabolic studies of cell suspensions is important in order to avoid artefacts in the determination of metabolite concentrations when using the spin-echo technique. Conversely it may be possible to perform transport measurements using spin-echo NMR to monitor the cell volume changes which occur during the transmembrane migration of molecules.     

Effect of physiological parameters of blood on magnetophoretic mobility of erythrocytes

Magnetophoresis is the process of the particle motion under the influence of a magnetic field. The magnetic particle and medium are considered responsive to the imposed magnetic field, and the material property that describes the response to the external magnetic field is relative magnetic permeability, and the magnetic susceptibility. The present work aims to evaluate the effect of internal and external physiological parameters on the erythrocytes' magnetophoretic mobility (MM) using Cell Tracking Velocimetry (CTV). The results of this study showed that there are a strong correlation between MM and several physiological blood parameters such as mean corpuscle hemoglobin (MCH), red blood cells distribution width (RDW), mean corpuscle hemoglobin concentration (MCHC), and fibrinogen.     

Effect of physiological parameters of blood on magnetophoretic mobility of erythrocytes

Magnetophoresis is the process of the particle motion under the influence of a magnetic field. The magnetic particle and medium are considered responsive to the imposed magnetic field, and the material property that describes the response to the external magnetic field is relative magnetic permeability, and the magnetic susceptibility. The present work aims to evaluate the effect of internal and external physiological parameters on the erythrocytes' magnetophoretic mobility (MM) using Cell Tracking Velocimetry (CTV). The results of this study showed that there are a strong correlation between MM and several physiological blood parameters such as mean corpuscle hemoglobin (MCH), red blood cells distribution width (RDW), mean corpuscle hemoglobin concentration (MCHC), and fibrinogen.     

Magnetic Susceptibility Difference-Induced Nucleus Positioning in Gradient Ultrahigh Magnetic Field

Despite the importance of magnetic properties of biological samples for biomagnetism and related fields, the exact magnetic susceptibilities of most biological samples in their physiological conditions are still unknown. Here we used superconducting quantum interferometer device to detect the magnetic properties of nonfixed, nondehydrated live cell and cellular fractions at a physiological temperature of 37°C (310 K). It is obvious that there are paramagnetic components within human nasopharyngeal carcinoma CNE-2Z cells. More importantly, the magnetic properties of the cytoplasm and nucleus are different. Although within a single cell, the magnetic susceptibility difference between cellular fractions (nucleus and cytoplasm) could only cause ∼41–130 pN forces to the nucleus by gradient ultrahigh magnetic fields of 13.1–23.5 T (92–160 T/m), these forces are enough to cause a relative position shift of the nucleus within the cell. This not only demonstrates the importance of magnetic susceptibility in the biological effects of magnetic field but also illustrates the potential application of high magnetic fields in biomedicine.     

Role of carbon substrates of culture media on Mycobacterium smegmatis susceptibility to antitubercular agents

Influence of medium composition on Mycobacterium smegmatis growth and susceptibility to antituberculosis drugs (ATD)--isoniazid and rifabutin--was studied. It was shown that addition of phospholipids (PL) in form of liposomes to meat peptone broth resulted in activation of M. smegmatis growth and decrease of its susceptibility to ATD. Growth characteristics of M. smegmatis and its susceptibility to ATD were studied using variants of modified on source of carbon synthetic medium Sauton as growth substrate. It was revealed that presence of acetate or PL in growth medium results in significant decrease of M. smegmatis susceptibility to isoniazid and rifabutin. It was suggested that this phenomenon is determined by activation of glyoxylate cycle by PL and fatty acids, which, in its turn, can stimulate expression of a number of proteins, including cell membrane pumps excreting antibiotics out of microbial cell.     

Magnetic susceptibility of iron in malaria-infected red blood cells

During intra-erythrocytic maturation, malaria parasites catabolize up to 80% of cellular haemoglobin. Haem is liberated inside the parasite and converted to haemozoin, preventing haem iron from participating in cell-damaging reactions. Several experimental techniques exploit the relatively large paramagnetic susceptibility of malaria-infected cells as a means of sorting cells or investigating haemoglobin degradation, but the source of the dramatic increase in cellular magnetic susceptibility during parasite growth has not been unequivocally determined. Plasmodium falciparum cultures were enriched using high-gradient magnetic fractionation columns and the magnetic susceptibility of cell contents was directly measured. The forms of haem iron in the erythrocytes were quantified spectroscopically. In the 3D7 laboratory strain, the parasites converted approximately 60% of host cell haemoglobin to haemozoin and this product was the primary source of the increase in cell magnetic susceptibility. Haemozoin iron was found to have a magnetic susceptibility of (11.0 ± 0.9) × 10^? 3 mL mol^? 1. The calculated volumetric magnetic susceptibility (SI units) of the magnetically enriched cells was (1.88 ± 0.60) × 10^? 6 relative to water while that of uninfected cells was not significantly different from water. Magnetic enrichment of parasitised cells can therefore be considered dependent primarily on the magnetic susceptibility of the parasitised cells.     

Red blood cell magnetophoresis

The existence of unpaired electrons in the four heme groups of deoxy and methemoglobin (metHb) gives these species paramagnetic properties as contrasted to the diamagnetic character of oxyhemoglobin. Based on the measured magnetic moments of hemoglobin and its compounds, and on the relatively high hemoglobin concentration of human erythrocytes, we hypothesized that differential migration of these cells was possible if exposed to a high magnetic field. With the development of a new technology, cell tracking velocimetry, we were able to measure the migration velocity of deoxygenated and metHb-containing erythrocytes, exposed to a mean magnetic field of 1.40 T and a mean gradient of 0.131 T/mm, in a process we call cell magnetophoresis. Our results show a similar magnetophoretic mobility of 3.86 x 10(-6) mm(3) s/kg for erythrocytes with 100% deoxygenated hemoglobin and 3.66 x 10(-6) mm(3) s/kg for erythrocytes containing 100% metHb. Oxygenated erythrocytes had a magnetophoretic mobility of from -0.2 x 10(-6) mm(3) s/kg to +0.30 x 10(-6) mm(3) s/kg, indicating a significant diamagnetic component relative to the suspension medium, in agreement with previous studies on the hemoglobin magnetic susceptibility. Magnetophoresis may open up an approach to characterize and separate cells for biochemical analysis based on intrinsic and extrinsic magnetic properties of biological macromolecules.     

Measurement of the susceptibility of paramagnetically labeled cells with paramagnetic solutions

A method of measuring the volumetric magnetic susceptibility, in which magnetically labeled cells or other particles are suspended in a paramagnetic solution of known susceptibility over the poles of a magnet, is presented. If the cells are more magnetic than the solution, they are attracted toward the poles; if they are less magnetic, they are repelled. If they have the same susceptibility as the solution, they do not move. Under this condition, the cells are said to be "isomagnetic" with the surrounding solution. Since the volumetric susceptibility of this solution is known, the susceptibility of the cells is obtained. Using the "isomagnetic" method, the volumetric susceptibilities of test metal powders were determined within +/- 8 X 10(-6) SI units. Yeast, colonic carcinoma, and liver cells, rendered magnetic with erbium chloride, had susceptibilities ranging from 13 to 20 X 10(-6). Particles of articular cartilage treated with erbium chloride were heterogeneous, with susceptibilities ranging between 50 and 125 X 10(-6), while particles of bone had a susceptibility of 560 to 580 X 10(-6). Eukaryotic cells labeled with ferritin attained susceptibilities of less than 1 X 10(-6).     

Effects of static magnetic fields at the cellular level

There have been few studies on the effects of static magnetic fields at the cellular level, compared to those of extremely low frequency magnetic fields. Past studies have shown that a static magnetic field alone does not have a lethal effect on the basic properties of cell growth and survival under normal culture conditions, regardless of the magnetic density. Most but not all studies have also suggested that a static magnetic field has no effect on changes in cell growth rate. It has also been shown that cell cycle distribution is not influenced by extremely strong static magnetic fields (up to a maximum of 10 T). A further area of interest is whether static magnetic fields cause DNA damage, which can be evaluated by determination of the frequency of micronucleus formation. The presence or absence of such micronuclei can confirm whether a particular treatment damages cellular DNA. This method has been used to confirm that a static magnetic field alone has no such effect. However, the frequency of micronucleus formation increases significantly when certain treatments (e.g., X-irradiation) are given prior to exposure to a 10 T static magnetic field. It has also been reported that treatment with trace amounts of ferrous ions in the cell culture medium and exposure to a static magnetic field increases DNA damage, which is detected using the comet assay. In addition, many studies have found a strong magnetic field that can induce orientation phenomena in cell culture.     

Quantification of cellular properties from external fields and resulting induced velocity: magnetic susceptibility.

An experimental technique is discussed in which the magnetic susceptibility of immunomagnetically labeled cells can be determined on a cell-by-cell basis. This technique is based on determining the magnetically induced velocity that an immunomagnetically labeled cell has in a well-defined magnetic energy gradient. This velocity is determined through the use of video recordings of microscopic images of cells moving in the magnetic energy gradient. These video images are then computer digitized and processed using a computer algorithm, cell tracking velocimetry, which allows larger numbers (>10(3)) of cells to be analyzed.     

Optimization of HIV-1 infectivity assays

HIV-1 reporter cell lines are the backbone of diagnostic assays, vaccine and drug development efforts. Performing HIV-1 infection experiments in a T cell background is desirable for many reasons. However, a low susceptibility to infection with primary patient isolates in available reporter T cell lines has limited such efforts. We here demonstrate that optimization of HIV-1 receptor expression and the utilization of serum free medium compositions can increase susceptibility of reporter T cell lines to HIV-1 infection by up to two orders of magnitude.     

Decrease in magnetic anisotropy of external segments of the retinal rods after a total photolysis.

The magnetic anisotropy of a polymembrane cell, such as the external segment of the frog retinal rod, is defined as the difference between the axial magnetic susceptibility (chiar), chia - chir = deltachi of the segment. After the total photolysis of the rhodopsin in situ, deltachi decreases a significantly by 20%. This decrease in magnetic anisotrophy should involve a subtle molecular disorder, mostly due to an alteration of the rhodopsin molecule.     

Biomolecular reactions studied using changes in Brownian rotation dynamics of magnetic particles

We have used magnetic particles to study specific binding of prostate specific antigen (PSA) to the surfaces of the bioparticles. The used particles have a mean diameter of about 130 nm and are placed in phosphate buffer saline (PBS). Each particle is composed of clusters of magnetic single domains of magnetite, which are covered with dextran. Changes in surface chemistry of the particles give rise to a change in the hydrodynamic volume of the particles. The later is mirrored by the changed frequency response of the complex magnetic susceptibility of a fluid containing these particles. Using ordinary induction coils and the lock-in amplifier technique it is possible to measure the complex magnetic susceptibility of the particle solution in a frequency range from about 10 Hz up to 10 kHz. From the measurement of the complex susceptibility versus the excitation frequency (both at the excitation frequency as well as at higher harmonics) we have shown that it is possible to quantitatively study the binding of PSA to the surfaces of the magnetic particles and thus to determine the PSA concentration in solution containing known concentration of nanoparticles functionalised with a monoclonal PSA antibody. Our method allows to perform an immunoassay in a single step and is much faster and cheaper compared to conventional ELISA procedures.