Stiffness of normal and pathological erythrocytes studied by means of atomic force microscopy.

During recent years, atomic force microscopy has become a powerful technique for studying the mechanical properties (such as stiffness, viscoelasticity, hardness and adhesion) of various biological materials. The unique combination of high-resolution imaging and operation in physiological environment made it useful in investigations of cell properties. In this work, the microscope was applied to measure the stiffness of human red blood cells (erythrocytes). Erythrocytes were attached to the poly-L-lysine-coated glass surface by fixation using 0.5% glutaraldehyde for 1 min. Different erythrocyte samples were studied: erythrocytes from patients with hemolytic anemias such as hereditary spherocytosis and glucose-6-phosphate-dehydrogenase deficiency patients with thalassemia, and patients with anisocytosis of various causes. The determined Young's modulus was compared with that obtained from measurements of erythrocytes from healthy subjects. The results showed that the Young's modulus of pathological erythrocytes was higher than in normal cells. Observed differences indicate possible changes in the organization of cell cytoskeleton associated with various diseases.

[1]  G. Semenza,et al.  Measuring elasticity of biological materials by atomic force microscopy , 1998, FEBS letters.

[2]  P. Wilairat,et al.  Severity differences in β‐thalassaemia/haemoglobin E syndromes: implication of genetic factors , 1993, British journal of haematology.

[3]  A. Ikai,et al.  MEASUREMENTS OF MECHANICAL PARAMETERS OF BIOLOGICAL STRUCTURES WITH ATOMIC FORCE MICROSCOPE , 1998 .

[4]  J. F. Greenleaf,et al.  Magnetic resonance elastography: Non-invasive mapping of tissue elasticity , 2001, Medical Image Anal..

[5]  J. Bereiter-Hahn,et al.  Cell cycle-related changes in F-actin distribution are correlated with glycolytic activity. , 1995, Experimental cell research.

[6]  P. Bolton-Maggs.  The diagnosis and management of hereditary spherocytosis. , 2000, Bailliere's best practice & research. Clinical haematology.

[7]  G. Watson,et al.  Imaging and force-distance analysis of human fibroblasts in vitro by atomic force microscopy. , 1999, Cytometry.

[8]  E. Evans,et al.  Mechanical properties of the red cell membrane in relation to molecular structure and genetic defects. , 1994, Annual review of biophysics and biomolecular structure.

[9]  Z Stachura,et al.  The effect of chitosan on stiffness and glycolytic activity of human bladder cells. , 2001, Biochimica et biophysica acta.

[10]  M. Langner,et al.  The kinetics of haemolysis of spherocytic erythrocytes. , 2003, Cellular & molecular biology letters.

[11]  V. Moy,et al.  Mechanical properties of L929 cells measured by atomic force microscopy: effects of anticytoskeletal drugs and membrane crosslinking. , 2006, Scanning.

[12]  M. Boogaerts,et al.  ADVERSE ROLE OF THE SPLEEN IN HEREDITARY SPHEROCYTOSIS: EVIDENCE BY THE USE OF THE ATOMIC FORCE MICROSCOPE , 1992, British journal of haematology.

[13]  I. N. Sneddon.  The relation between load and penetration in the axisymmetric boussinesq problem for a punch of arbitrary profile , 1965 .

[14]  H. Haga,et al.  Elasticity mapping of living fibroblasts by AFM and immunofluorescence observation of the cytoskeleton. , 2000, Ultramicroscopy.

[15]  M. Bibb,et al.  Engineering of Primary Carbon Metabolism for Improved Antibiotic Production in Streptomyces lividans , 2002, Applied and Environmental Microbiology.

[16]  Z. Stachura,et al.  Elasticity of normal and cancerous human bladder cells studied by scanning force microscopy , 1999, European Biophysics Journal.

[17]  M. Schaller.  Biochemical signals and biological responses elicited by the focal adhesion kinase. , 2001, Biochimica et biophysica acta.

[18]  Y. Ravindranath,et al.  Oxidant Damage to Erythrocyte Membrane in Glucose-6-Phosphate Dehydrogenase Deficiency: Correlation With In Vivo Reduced Glutathione Concentration and Membrane Protein Oxidation , 1994 .

[19]  N. Wijayanti,et al.  Biology of heme in health and disease. , 2004, Current medicinal chemistry.

[20]  W. Kalow.  GENETIC FACTORS IN RELATION TO DRUGS. , 1965, Annual review of pharmacology.

[21]  Joan M. Lau,et al.  Atomic force microscopy imaging of living cells: a preliminary study of the disruptive effect of the cantilever tip on cell morphology. , 2000, Ultramicroscopy.

[22]  P. Luckham,et al.  Imaging erythrocytes under physiological conditions by atomic force microscopy. , 2001, Biochimica et biophysica acta.