Oral 23rd International Society of Magnetic Resonance Conference 2023

Cell morphing sensed with multiple-quantum filtered 23Na NMR (#22)

Thomas R Eykyn 1 , Stuart J Elliott 2 , Philip W Kuchel 3
  1. School of Biomedical Engineering and Imaging Sciences, King's College London, St Thomas' Hospital, London SE1 7EH, United Kingdom
  2. Molecular Sciences Research Hub, Imperial College London, London W12 0BZ, United Kingdom
  3. School of Life and Environmental Sciences, The University of Sydney, Sydney, New South Wales, Australia

Red blood cells (RBCs) undergo an autonomous/spontaneous shape change when depleted of glucose or are poisoned with various reagents, including 20 mM NaF. They change from their normal biconcave disc shape (discocyte [1]), to a spiky spherical form (echinocyte), and then on to a sphere (spherocyte), over periods of 10’s of minutes. This shape evolution is called the DEST morphological transformation. Because discocytes align with their flat(ish) faces parallel to B0 in an NMR magnet [2], the average anisotropy of the electric field gradients (EFGs) associated with the cell membrane leads to an enhanced double-quantum filtered (DQF) 23Na+ NMR signal from the ions, especially outside the cell. When the cell shape changes the EFGs becomes progressively spherically averaged, so the DQF declines. A time course of the DQF spectral intensity correlates with an increase in echinocyte and spherocyte numbers in images recorded with differential interference contrast (DIC) light microscopy. Various experimental developments led to control of the rate of the DEST. These will be explained, along with the theoretical basis of the observations that rely on quantum mechanical theory of irreducible spherical tensors [3,4]. Cross-correlation between quadrupolar- and paramagnetic-relaxation mechanisms/pathways means that conversion of haemoglobin to the highly paramagnetic FeIII -containing form (methaemoglobin) leads to emergence of a DQF-signal from 23Na+ even in isotropic haemolysates (RBCs disrupted by freezing and thawing). In other words, a DQF 23Na+ NMR signal from RBCs that have been biochemically manipulated allows the population size of such altered cells to be determined from mixtures with normal RBCs. The rationale for wanting to do this in studies of mechanosensation in RBCs will be explained.

  1. Kuchel, P. W., Cox, C. D., Daners, D., Shishmarev, D., and Galvosas, P. (2021) Surface model of the human red blood cell simulating changes in membrane curvature under strain. Sci. Rep. 11, 13712, 18.
  2. Kuchel, P. W., Coy, A., and Stilbs, P. (1997) NMR “Diffusion-diffraction” of water revealing alignment of erythrocytes in a magnetic field and their dimensions and membrane transport characteristics. Magn. Reson. Med. 37, 637.
  3. Shinar, H., Knubovets, T., Eliav, U., and Navon, G. (1993) Sodium interaction with ordered structures in mammalian red blood cells detected by Na-23 double quantum NMR. Biophys. J. 64, 1273.
  4. Knubovets, T., Shinar, H., Eliav, U., and Navon, G. (1996) A 23Na multiple quantum-filtered NMR study of the effect of the cytoskeleton conformation on the anisotropic motion of sodium ions in red blood cells. J. Magn. Reson. B 110, 16-25.
  5. Ling, W., and Jerschow, A. (2007) Relaxation-allowed nuclear magnetic resonance transitions by interference between the quadrupolar coupling and the paramagnetic interaction. J. Chem. Phys. 126, 064502, 1-6.