Red blood cell rheology using single controlled laser-induced cavitation bubbles.

The deformability of red blood cells (RBCs) is an important property that allows the cells to squeeze through small capillary vessels and can be used as an indicator for disease. We present a microfluidic based technique to quantify the deformability of RBCs by stretching a collection of RBCs on a timescale of tens of microseconds in a microfluidic chamber. This confinement constrains the motion of the cell to the imaging plane of the microscope during a transient cavitation bubble event generated with a focused and pulsed laser. We record and analyze the shape recovery of the cells with a high-speed camera and obtain a power law in time, consistent with other dynamic rheological results of RBCs. The extracted exponents are used to characterize the elastic properties of the cells. We obtain statistically significant differences of the exponents between populations of untreated RBCs and RBCs treated with two different reagents: neuraminidase reduces the cell rigidity, while wheat germ agglutinin stiffens the cell confirming previous experiments. This cavitation based technique is a candidate for high-throughput screening of elastic cell properties because many cells can be probed simultaneously in situ, thus with no pre-treatment.

[1]  E. Sackmann,et al.  Viscoelastic properties of erythrocyte membranes in high-frequency electric fields , 1984, Nature.

[2]  Jerry L. Prince,et al.  Snakes, shapes, and gradient vector flow , 1998, IEEE Trans. Image Process..

[3]  G. Seaman,et al.  Sialic Acids and the Electrokinetic Charge of the Human Erythrocyte , 1961, Nature.

[4]  Subra Suresh,et al.  Viscoelasticity of the human red blood cell , 2006, American journal of physiology. Cell physiology.

[5]  Claus-Dieter Ohl,et al.  Sonoporation of suspension cells with a single cavitation bubble in a microfluidic confinement. , 2007, Lab on a chip.

[6]  Shamik Sen,et al.  Indentation and adhesive probing of a cell membrane with AFM: theoretical model and experiments. , 2005, Biophysical journal.

[7]  N. Mohandas Molecular basis for red cell membrane viscoelastic properties. , 1992, Biochemical Society transactions.

[8]  D. Aminoff The role of sialoglycoconjugates in the aging and sequestration of red cells from circulation. , 1988, Blood cells.

[9]  H. Kataoka,et al.  Dynamic deformation and recovery response of red blood cells to a cyclically reversing shear flow: Effects of frequency of cyclically reversing shear flow and shear stress level. , 2006, Biophysical journal.

[10]  Nico de Jong,et al.  Sonoporation from jetting cavitation bubbles. , 2006, Biophysical journal.

[11]  E. Evans,et al.  New membrane concept applied to the analysis of fluid shear- and micropipette-deformed red blood cells. , 1973, Biophysical journal.

[12]  D. Discher,et al.  Power-law rheology of isolated nuclei with deformation mapping of nuclear substructures. , 2005, Biophysical journal.

[13]  W. Huestis,et al.  Wheat germ agglutinin stabilization of erythrocyte shape: role of bilayer balance and the membrane skeleton. , 1995, Biochimica et biophysica acta.

[14]  D. Branton,et al.  Identification of the Protein 4.1 Binding Interface on Glycophorin C and p55, a Homologue of the Drosophila discs-large Tumor Suppressor Protein (*) , 1995, The Journal of Biological Chemistry.

[15]  Subra Suresh,et al.  Large deformation of living cells using laser traps , 2004 .

[16]  R M Hochmuth,et al.  Membrane viscoelasticity. , 1976, Biophysical journal.

[17]  D. Branton,et al.  In vitro binding studies suggest a membrane-associated complex between erythroid p55, protein 4.1, and glycophorin C. , 1994, The Journal of biological chemistry.

[18]  E. Evans,et al.  Adhesivity and rigidity of erythrocyte membrane in relation to wheat germ agglutinin binding , 1984, The Journal of cell biology.

[19]  R. Hochmuth,et al.  Red cell extensional recovery and the determination of membrane viscosity. , 1979, Biophysical journal.

[20]  G. J. Brakenhoff,et al.  A NEW METHOD TO STUDY SHAPE RECOVERY OF RED BLOOD CELLS USING MULTIPLE OPTICAL TRAPPING , 1995 .

[21]  J. Dormandy,et al.  Abnormalities in the mechanical properties of red blood cells caused by Plasmodium falciparum , 1989 .

[22]  Claus-Dieter Ohl,et al.  Interaction between two laser-induced cavitation bubbles in a quasi-two-dimensional geometry , 2009, Journal of Fluid Mechanics.

[23]  Claus-Dieter Ohl,et al.  Controlled cavitation in microfluidic systems. , 2007, Physical review letters.

[24]  R. Simmons,et al.  Elasticity of the red cell membrane and its relation to hemolytic disorders: an optical tweezers study. , 1999, Biophysical journal.

[25]  J. Conboy,et al.  Glycophorin C content of human erythrocyte membrane is regulated by protein 4.1. , 1990 .

[26]  N. Mohandas,et al.  Red blood cell deformability, membrane material properties and shape: regulation by transmembrane, skeletal and cytosolic proteins and lipids. , 1993, Seminars in hematology.

[27]  D. W. Knowles,et al.  Cooperative action between band 3 and glycophorin A in human erythrocytes: immobilization of band 3 induced by antibodies to glycophorin A. , 1994, Biophysical journal.

[28]  Shu Chien,et al.  Role of Surface Electric Charge in Red Blood Cell Interactions , 1973, The Journal of general physiology.

[29]  M W Berns,et al.  Laser-micropipet combination for single-cell analysis. , 1998, Analytical chemistry.

[30]  J. Käs,et al.  The optical stretcher: a novel laser tool to micromanipulate cells. , 2001, Biophysical journal.

[31]  J. Conboy,et al.  Regulation of Protein 4.1R, p55, and Glycophorin C Ternary Complex in Human Erythrocyte Membrane* , 2000, The Journal of Biological Chemistry.

[32]  R Vanderby,et al.  Interrelation of creep and relaxation: a modeling approach for ligaments. , 1999, Journal of biomechanical engineering.

[33]  Vasan Venugopalan,et al.  Laser-induced mixing in microfluidic channels. , 2007, Analytical chemistry.

[34]  W. Yao,et al.  Influence of neuraminidase on the characteristics of microrheology of red blood cells. , 2000, Clinical hemorheology and microcirculation.

[35]  M. Arpin,et al.  Membrane-actin microfilament connections: an increasing diversity of players related to band 4.1. , 1994, Current opinion in cell biology.

[36]  N. Mohandas,et al.  Modulation of Band 3-Ankyrin Interaction by Protein 4.1 , 1996, The Journal of Biological Chemistry.

[37]  A Leung,et al.  Static and dynamic rigidities of normal and sickle erythrocytes. Major influence of cell hemoglobin concentration. , 1984, The Journal of clinical investigation.

[38]  Maggs,et al.  Subdiffusion and Anomalous Local Viscoelasticity in Actin Networks. , 1996, Physical review letters.

[39]  Vasan Venugopalan,et al.  Pulsed laser microbeam-induced cell lysis: time-resolved imaging and analysis of hydrodynamic effects. , 2006, Biophysical journal.

[40]  Vasan Venugopalan,et al.  Examination of laser microbeam cell lysis in a PDMS microfluidic channel using time-resolved imaging. , 2008, Lab on a chip.

[41]  S. Suresh,et al.  Cell and molecular mechanics of biological materials , 2003, Nature materials.