Cell volume dependence of 1H spin-echo NMR signals in human erythrocyte suspensions: The influence of in situ field gradients |
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Authors: | Zoltan H. Endre Philip W. Kuchel Bogdan E. Chapman |
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Affiliation: | Department of Biochemistry, University of Sydney, Sydney, New South Wales, 2006 Australia |
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Abstract: | 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. |
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Keywords: | Spin echo Cell volume Field gradient (Human erythrocyte) A signal amplitude volume magnetic susceptibility translational diffusion coefficient Δδ chemical shift difference fraction of osmotically inactive cell water f.i.d. free induction delay mean field gradient GSH glutathione NMR nuclear magnetic resonance π osmotic pressure isotonic osmotic pressure τ delay time spin-spin relaxation time TMS tetramethylsilane erythrocyte volume erythrocyte volume at isotonicity |
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