Electric field induced instabilities at bilayer membranes and fluid-fluid interfaces Rochish Thaokar Department of Chemical Engineering IIT Bombay, Mumbai (Bombay), India 25th May 2012, KITPC, Beijing, China 1 Outline Rayleigh Plateau Instability in Fluid Jets Brief Introduction to Pearling in Cylindrical vesicles Brief Introduction to Electrostatics and Electrohydrodynamics Pearling under Uniform electric fields Conclusions Fluid-Fluid Electrohydrodynamics: Planar interfaces and drops 2 Apology The talk is a little more elaborate version of my short talk The fluid-fluid part is new, would like to have your inputs on making connection with bio-physics 3 Rayleigh-Plateau Instability: Basic Physics Total Energy=Surface tension*area Long wave perturbations reduce a jet’s area Instability happens when the wavelength of the perturbation is larger than the circumference of the cylinder r=1+D ei(kz+mθ)+st) Pin =γ(1-δ D(1-m2-k2)e i(kz+mθ)+st) Normal mode analysis yields a simple kinematic explanation for the instability. Stabilizing longintudinal curvature and destabilizing azimuthal curvature 4 Which wavenumber is unstable? Seen in experiments Pin =γ(1-δ D(1-m2-k2)e i(kz+mθ)+st) S=Ak(1-k^2) km Long wave instability (k<1 or λ>2 π R) are unstable Rayleigh’s analysis Medium Air: Viscous (km=0) Inviscid (km=0.7) Tomotika (Both fluids viscous) (km=0.56) 5 Rayleigh Plateau in cylindrical vesicles (Pearling) In most simple cylindrical vesicular systems, the tension is identically zero. (Tension due to thermal fluctuations too weak to induce instability). Cylindrical vesicles do not pearl on their own. The tension required for pearling is c 3 B 2a 2 Bar Ziv and Moses, 1994, PRL, first showed Laser Induced pearling Transfer of energy of Laser results in a tension in the membrane that causes pearling (Dielectrophoretic effect) 6 What could be issues in RP in vesicles Fluid jets can decrease their area (area is not conserved) In cylindrical vesicles, the membrane area has to be conserved RP instability leads to reduction of area. A tense vesicle would have to displace the reduced area This is unlike planar membrane analysis where area is pulled in The process of deflection of area can also lead to front velocity 7 A brief about cylindrical vesicles HB B 2 ( 2 H ) dA dA ( P Bending Area 2 e For a sphere of radius a Minimise HB: Pi ) dV volume conservation Pi Pe 2 Hbend=8 πκB a Pressure independent of bending modulus, Energy independent of radius For a cylinder, minimise wrt a and L Pi Pe 2 B a 3 Pi Pe 3 B 2a 2 Pi Pe Pe Pi a 2 a B 2a 3 B a 3 Pe Pi Negative pressure contribution by the bending term When length does not matter (very long cylinders), Pi= Pe and at equilibrium a=√κ/2σ, external tension 8 Stretched cylindrical vesicles HB B 2 ( 2 H ) dA dA ( P Bending Area 2 e Pi ) dV -f L volume conservation f f For a cylinder, minimise wrt a and L, without the volume constraint (?), infinite reservoir of fluid outside a=√κ/2σ f=2 π √2 κσ =2 π κ/a In synthesis though, the radius is decided by the preparation conditions When stretched, there might be dynamics associated reduction to equilibrium radius a = √κ/2σ (viscosity controlled) 9 Salient observations in Bar-Ziv et. al’s work (PRL 1994) No intrinsic curvature, no initial tension Far from equilibrium system (Slow dynamics) Laser 50mW with 0.3 microns radius, generates tension of 1.8 10-4 mN/m. Lipid molecules sucked into the laser (akin negative dielectrophoresis) A wavenumber k=2πR/λ=0.8-1.0 of the instability is observed Significantly different from the fluid-jet analysis 10 Salient observations in Bar-Ziv et. al’s work The reduced area during RP instability is absorbed in the laser trap Leads to a propogating instability from the laser trap A front seen to propogate at around 30 microns/s This velocity increases with laser power, tension 11 Late stage pearling The reduced volume in a cylinder in the large L limit is v=3/21/3 (R/L)1/2 • Large L leads to small v can have variety of equilibrium shapes!! • Late stage pearling!! Volume conservation leads to, Rp=1.806 Ro Rneck= √κ/2σ=470 nm 12 Different techniques for inducing Pearling Instability in cylindrical vesicles Optical Tweezers (Bar-Ziv et al., 1994) Polymer anchoring (Tsafrir, 2001) Magnetic field (Menager et al., 2002) Elongational flow (Kanstler, 2008) Nanoparticle encapsulation (Yan Yu and Steve ,2009) Application of tweezers on membrane creates surface tension by drawing lipid molecules into the tweezed area Spontaneous curvature because of the amphiphilic polymer backbone induces tension in the outer leaflet of bilayer membrane tube Deformation of magnetoliposome takes place under applied magnetic field leading to tension in the cylinder Stretching of a tubular vesicles with initial length to dia. ratio L/D0 > 4.2 by an elongational induces shape transformation from dumbbell to a transient pearling state Encapsulation of excess of nanoparticles within GUVs induces 13 shape transformation Synthesis of cylindrical vesicles Spin coating (1kRPM, 10sec) of microscopic glass slide with DMPC lipid SS-electrodes (Thickness 0.45mm) at a spacing of 3mm DMPC Lipid Conc. 1,2Dimyristoylsn-glycero-3phosphocholin e 10 mg/ml (CHCl3:CH3 OH = 2:1) Spin coating 1000 RPM for 10 sec Sucrose solution conc. 0.1M Electrode thickness 0.45mm (Stainless steel) Electrode spacing 3mm Conductivity (0.1M Sucrose solution) 5.7µS/cm Sucrose solution injection rate 3ml/min Fixing upper glass slide to the bottom one Sealing from four sides to form a closed chamber Dry lipid layer hydration by sucrose solution injection (3ml/min) All the experiments conducted at 26 oC above Tg(23 oC) 14 Images of Cylindrical vesicles Variety of sizes of cylindrical vesicles observed Vesicles appear as single cylinders or a bunch They are free at both ends or connected to lipid reservoirs Myelin and multi-lamellar cylindrical vesicles also observed 15 Electric field Experimental setup Computer CCD Camera Inverted microscope Experimental cell Figure: Electric field setup Function generator High frequency amplifier 16 2.5 mm spacing DC experiments without amplifier (Voltages around 1.5 V) AC experiments: 500kHz to 2 MHz (Voltages around 60 V) Oscilloscope Some important experimental observations Pearling Late Pearls Some important experimental observations Flutter Budding 18 Some important experimental observations Pearling seen on increasing the field Seems to start at one end of the cylinder Late pearls in some cases show bimodal distribution Simultaneous stretching is observed but is remarkably reversible Flutter at strong fields. A fluttered vesicle often pearled on removal of field: Tension is dissipated much slowly μea/σ 19 Effect of electric field In most systems, the tension is almost zero: Cylindrical vesicles do not pearl on their own!! The tension required for pearling is c 3 B 2a 2 How does electric field induce tension in a cylindrical vesicle? Problem complicated by end-caps. What is the field distribution around end caps? Axial part End Caps No Simple base state on which stability analysis can be conducted Normal mode analysis difficult if ends are considered 20 Maxwell’s stress (Origin) E + + + + - Net Maxwells force due to difference + + in Dielectric constants 1 2 + =ρ E-1/2 εo E.E grad ε =del.T Τ=F/Area=ε εo (EE-1/2 I E2) E -- ++ + + F/Vol=ρ E+P.grad E-1/2 grad εo(ε -1) E.E I + Net Maxwells force due to difference in conductivities + Air - - + E cos(ωt) + 21 What are the axial and end-cap forces? Consider the vesicle to be a dielectric in a conductor medium Helfrich (1983) in DC fields. Eo Axial part n . e e o ( EE n E 2 2 n ) i o ( EE n E 2 2 n ) e o Eo 2 n 2 Compressive axial electric stress on the walls What about the caps? Solve for electrostatics on a spherical vesicle, and consider one half of the same 22 End Caps: Electrostatics for a spherical vesicle m ,e 0 2 •Assume spherical vesicle as a dielectric drop in a conducting liquid (Helfrich 1983) 2 a=6 microns m •Continuity of potential at interface of membrane inner and outer medium •Normal field zero at the conductor-dielectric interface n.E e 0 ( r a ) e m (r a ) •Compressive stresses 9 E o a e o 2 F E DC 2 16 23 e Can axial and end-cap compressive forces generate tension? How is tension generated by compressive electric stresses? Eo Axial part n . e e o ( EE n E 2 2 n ) i o ( EE n E 2 2 n ) e o Eo 2 n 2 Compressive axial electric stress F c 9 E o a e o 2 E DC 2 16 3 B 2a 2 24 Effect of membrane thickness on electric field Electric field calculations assuming the vesicle to be a dielectric drop incorrect, although one can still predict generation of tension The membrane is just a thin layer of dielectric. The inner core is a conductor and although the field inside is zero, the charges at the inner core would be substantial A detailed model to describe the electrostatics should be suggested. The net electric traction is f E n . e ( r a d ) n . m ( r a d ) n . m ( r a ) n . i ( r a ) d=5nm 2 a=6 microns i e m 25 Dielectrics, Leaky dielectrics and Conductors Perfect Dielectrics + 1 2 -- ++ + Layered Dielectrics (PD-PD): Net bound charge at the interface E - - + + + +- +- + + Leaky Dielectrics + + + -- - + + ++ + - ++ + E Layered Dielectrics (LD-LD): Accumulation of charges at the interface. The charge relaxation time is given by tc=ε/σ 1 E1 2 E 2 Conductors Steady state assumption in most cases Charges accumulate at the interface Assumption that charge relaxation time Equi-potential assumption tR=ε/σ(t is faster than other time-scales (Low frequency) E E 1 1 2 2 Current continuity condition Is realised when the conductivity is very large High frequency: Dielectrophoretic behaviour 26 1 E1 2 E 2 Electrostatics equations i ,m ,e 0 2 •No free charge, conductors, perfect or leaky dielectrics 2 a=6 microns •The boundary conditions are important i m •Continuity of potential at interface of membrane inner and outer medium o ( e E e m E m ) q e ( r a d ) m Em eEe o ( m E m i E i ) q i ( r a ) i Ei m Em qe t qi t (r a d ) (σi=σe=5 10-5 S/m σm=0 εe=εi =80 εm =2) (r a ) Typically, we assume σm=0 tMW=εe εo /σ e e o (qe m E m ) ( q e m E m ) t MW qe t qe t Conductor Behavior ω>t-1MW Dielectric behavior ω<t-1MW Water (5 10-5 S/m) , t-1MW =70 kHz 27 e Model 2 f E n . e ( r a d ) n . m ( r a d ) n . m ( r a ) n . i ( r a ) DC Case ω<<tmw-1 High frequency ω>tmw-1 Model 1 9 a E o m o 3 F E DC 2 8d Tensile Axial stress 2 2 a F E Ec obtained by requiring F E AC /2 0 adE 2 o ( e m ) o 2 2 m d f E a sin c 2 3 B 2a 2 28 Critical Electric field for pearling Vesicles turn in the direction of field The frequency dependent tension, when exceeds the critical tension, onset of Pearling is observed Both AC and DC experiments are reported Low DC voltage and fields to prevent electroporation (<1kV/cm, DC) and electrolysis 29 Governing equations and Boundary conditions Variables Scalings X a T μea3/κB For Hydrodynamics, membrane acts as an interface V κB /μea2 Electrostatics solved for internal and external fluids and the membrane phase P,τ κB / a3 Φ,E √κB /aεo, √κB /a3εo ω κB/μea3 Linear stability analysis is conducted Stokes equations for Hydrodynamics 30 Stability Analysis Put normal mode perturbations for all the variables Get dispersion relation and determine the value of s m=0 is the symmetric mode m=1,2 are the non-axisymmetric modes Low wave number instability is often seen Floquet analysis is conducted for time-periodic voltages 31 Rayleigh Plateau instability in liquid-liquid jets For εi > εe, the Maxwell’s stress is out of phase with the displacement D by Π/2, stabilizing action of the electric field. At B the field is obstructed so +ve free charges -ve perturbation charges at position A. Axial perturbation electric field e is in phase with the interface displacement D. E-Field stabilizes RP instability in liquid-liquid jets Base state stress at interface is -(εe -εi)/2 E2 and is compressive The normal perturbation stress is -(εe -εi)/2 e E and is directed inwards at the crests and outwards at the trough, leading to stabilization. e.g electrospinning 32 Rayleigh Plateau instability in cylindrical vesicles Governing equations and Boundary conditions Normal stress BC has a tension term Intrinsic tension due to electric field (Maxwell’s stresses, in the base state) Perturbed stress, incompressibility condition leads to a tension (a Lagrange parameter) Compare with fluid-fluid model (No tension, tangential stress continuity) or immobile interface (zero tangential velocity) 33 Effect of electric field on wavelength Dual Role of Electric field: It generates tension needed to induce the instability (σ> σc). But also suppresses RP instability in jets Balance of electric field induced tension and stabilizing effect of E yeilds an E independent plateau km This results in increase in km with E and plateaus to a value less than 0.56 (fluids) DC fields: Two possible cases (Ee =Eo , Em =Ei=0) and (Ee =Em =Ei=Eo) The plateau value of km decreases with an increase in the frequency 34 Effect of electric field on wavelength: Comparisons with Experiments DC Experiments AC Experiments Laser Tweezing Data (BarZiv and Moses, PRL 1994) Issues: Issues: Significant scatter in the than km theory much smaller experimental Experiments data The fields DC Either MSCrequired effect orfor some instability much smaller than AC Physics missing? Weak dependence of electric field is seen unlike fluid jets 35 Conclusions Late stage Pearl size (When unimodal distribution) Rp=1.806 Ro Completely reversible Pearling instability observed and explained Dual behavior of Electric field: Induces tension as well as stabilizes the instability Experimental km values significantly higher than the theory Analytical theory does not predict flutter if membrane is non-conducting 36 Research in my group Electrohydrodynamics in fluid-fluid systems Recently started working in the area of “Effect of fields on bilayers and vesicles (Spherical and cylindrical)” Apologies for the fluid-fluid systems. Would be keen to know if there are similar problems of biological importance 37 Electrohydrodynamic instabilities at fluid-fluid interfaces in low conductivity, low frequency limit Possible Mechanism of Electroformation System GOAL Shimanouchi, Langmuir, 25(9),2009 Y=βho external fluid (e) internal fluid (i) Bilayer (m) Late stage swelling under osmosis and maxwell stresses Y=0 Y=- ho Fluid 1 Fluid 2 ho λ WE ARE HERE 39 Introduction Soft Pattern Lithography replication is an important tool in many industries. More challenging for lower length scales. Methods Photolithography Advantages Sizes accessible: 50nm Electronbeam lithography Disadvantages: Complex chemical treatments, needs a Direct positive replicaclean of the mask. mask, needs room. Sizes accessible: <10nm Soft Lithography Inexpensive Complex and expensive Fig:Disadvantages: a) PET preform mould b) Pressure driven microfluidic instrumentation bioreactor 40 No sophisticated devices/ expertise required Schaffer al, 2000, Morariu et al, 2002, Deshpande et al, 2004 Website: et blowmolding-machine.en.made-in-china.com and loac-hsg-imt.de Soft Lithography: The story so far 1999 Chou et al 2001 Schaffer et al, Lin et al 2002-04 Morariu et al, Leach et al 2006-08 Dickey et al, Voicu et al 2009-10 41 Heier et al, Sharma et al • LISA (no electric fields), Pattern depends on mol weight of the polymer • Electric fields , hexagonal pattern, rosette structure, linear mask replicated, bilayers • Patterned electrodes, polymerpolymer-air trilayers • Novel 3d cages, high aspect ratio pillars, time evolution of instability • Spatially Modulated fields • Viscous to elastic behaviour Experiments (Protocol) Scotch tape is stuck on the lower slide as spacer A drop of Poly dimethyl siloxane 30000cSTk fluid is placed The fluid is spin coated at 3k rpm for 3 mins to get a ~37 micron film The second slide is placed against the first and it is connected to AC supply Undulations form and grow at the fluid interface to form columns which touch the top slide. The mean spacing is then measured a Pattern formation: Simple theory 2 e o (V / H ) s H / a b1 / 2 1 / 2 a c E H 3/ 2 V 2 1 b c500 Hz 1 kV/cm d c 500 Hz 0.5 kV/cm DC 0.5 kV/cm d DC 0.5 1 kV/cm Ω 43 Mechanism of the instability 44 1 2 ε1 E + - - + + + + + + + + + + + + ε2 1 2 ε1 High pressure in fluid 1 E + - - + + + + + + + + + + + + ε2 Net negative bound charge Base State 1 2 m1 ε1 y E ε2 m 2 y 1 2 ε1 ~ ~ 2 E 1 Point of equal potential ~ 1 ~ 2 ε2 1 2 + ε1 + - - - - + + + + + E Net negative bound perturbation charge ε2 - + + + + Net positive bound perturbation charges 1 2 ε1 E ε2 Leads to attraction Mechanism of the Instability (leaky dielectrics) 1 2 + 12 + - - - + + - - + + + - + - - ε1 + - + - ++ - + + + - + -+ ε2 1 2 m1 12 σ1 y Different conductivities of the two fluids results in this base state For the case of equal ε m 2 y σ2 1 2 σ1 + 12 - - + + -q + - - + +- + σ2 Leads to a base state charge given by q=ε1E1-ε2E2 + + + - + -+ 1 2 σ1 12 ~ ~ 2 E 1 Point of equal potential σ2 1 2 σ1 12 - + +- - - - +- - + E Net negative perturbation charge σ2 1 2 12 ε1 E ε2 Leads to attraction Model Y=βho Fluid 1 ho Y=0 Y=- ho Fluid 2 λ 57 Model Boundary Conditions Governing Equations Continuity of velocities u 1 u 2 Equation of continuity Balance of normal and tangential stresses .v 0 n.( 1 2 ). n n.( Momentum Balance v t e 2 ). n gh ( x , t ) e 1 n .( 1 2 ). t n .( ~0 Re( v . v ) p v 2 Gauss’s law for electrostatic potential 0 2 No free charges and electric body force in the bulk v1 v 2 Kinematic condition e 1 h t e 2 ). t 0 V . s h v 2 Continuity of potential in normal and tangential direction 1 2 0 1 1 y 0 2 2 y q Charge conservation equation q t u . s q qn .( n . ) u 1 1 y 2 58 2 y Electric stress Scaling Parameter General model Thin Film approx. Length h0 h0 0 0 Velocity 2 2 h0 0 0 2 Pressure h0 Time 2 h0 0 0 Interfacial Charge Conductivity 2 2 or for AC 2 h0 2 0 0 2 h0 h0 2 h0 2 2 1 or 2 0 0 2 2 h0 2 h0 2 0 0 for AC 1 2 A small parameter 0 0 2 h0 s 3 2 2 lateral length scale 0 0 2 0 0 0 0 2 2 h0 e Stress due to surface tension 0 0 2 2 h0 0 0 2 h0 2 1 h0 0 0 h 0 2 h0 0 0 h 0 2 1 2 Tools used LSA for AC and DC systems Nonlinear simulations using thin film approximation Comparison with experiments 60 Effect of conductivity in DC experiments Linear Stability Analysis Perturbations are expressed as ikx st ~ m m0 me k- wavenumber of the instability s- growth rate of the instability Instability is characterized by a fastest growing mode (kmax) kmax grows = k1 + + k2 k3 The inverse of kmax gives the wavelength of the pattern obtained experimentally. 62 Background Two time scales of interest Time for growth of instability (τs=1/smax) Time for charge migration/relaxation (τc =ε/σ) If τc >> τs ---- PD-PD If τc << τs ---- PD-Conductor If τc ~ τs ---- PD-LD Linear Stability analysis assumes a well defined base state (A conductor) What do you compare experiments? SIMULATIONS Background Case : Both Charge Instability happen relaxes grows simulatneously faster beforethan charge the instability migrates grows A leaky-leaky interface c cc s s ? In this case, interfacial charge doesn’t reach its steady state value and the A perfect dielectric leaky dielectric behaviour assumption of the linear theory becomes invalid. Non-linear simulations are requiredbehaviour 64 Results for DC fields simulation The actual wavelength seen in experiments decreases considerably with decrease in the conductivity as predicted by the non-linear 65 analysis. AC Conditions Important questions? Effect of frequency of AC fields on wavelength of patterns: Theory and Experiments? (Not reported yet) Frequency as a tool (identical to rheometry) to probe different time scales Can simulations reveal more about instabilities under AC fields? τc << τs and τω< τs (Fully charged surface) τω<< τc and τω >> τc For τc << τs and τω> τs For τc =τs and τω> τs and τω< τs 67 Experiments c s and s c s and s ND conductivities are 20, 1 and 0.05 c s and s Experiments (Scale frequency with conductivity) 69 Observations No fitting parameters Reasons for disagreement : Errors in conductivity measurement High polydispersity in the pattern, especially at high frequencies (low growth rates) Beta, the ratio of heights of air and liquid is not the same in all the experiments. 70 Conclusions The non-dimensional conductivity does matter in electrohydrodynamic instabilities for both AC and DC fields Simulations might be necessary to make accurate predictions The experimental observations support the above two 71 Acknowledgements DST Priya Gambhire, PhD student 72 Drop deformation and translation in non-uniform fields Why nonuniform elecric field Drops deform and break or coalesce under electric fields What is the best electrode configuration? Dielectrophoresis is movement of a particle in non-uniform fields Dielectrophoresis has several applications biophysics, bioengineering, multiphase separation To investigate drop deformation, breakup and motion in the simplest non-uniform electric field 74 Why nonuniform elecric field Drop Transport by Dielectrophoresis Breakup of water drop in castor oil Nature, 426(2003) 515 Separation of Tobacco Mosaic Virus(+ve dielectrophoresis) and Herpes Simplex Virus(-ve dielectrophoresis) (Kua C. H. et al 2004) Deformation, dielectrophoresis and oscillation of water drop in castor oil 75 Drop breakup under P1 Electric field Large CaE LD systems showing breakup with prolate deformation Q=10, R=0.1,CaE=0.342 Ca E R Large CaE LD systems showing breakup with oblate deformation (Q=10, R=10,CaE=0.342) 0 E 2 a Q Summary of Leaky dielectric results R Q<1 No deformation for quadrupole field No deformation for uniform field No electrohydrodynamic flow λ = 0.01 R Q>1 λ =1 λ = 100 ··· — --- Salient features: Boundaries for prolate and oblate changed Possibly the drop shapes are also more complex 77 Acknowledgements Shivraj Deshmukh Sameer Mhatre Thank You 78 Planar Membranes HB B 2 (2 H ) 2 dA dA ( P e h Pi ) dV Shape equation • For the planar case, the height-height correlation of a fluctuating membra assumes no area conservation!! Membrane can be drawn from the sides. • One can still enforce local lipid conservation. • In 1-D it means the tangential velocity is zero • In linearised theory, the tension is O(ε) • • The tension γ is always externally imposed ( hq hq * ) kT A ( q q 4 ) 2 It is not the tension (Lagrange Multiplier) to conserve area 79 Planar Membranes: Questions • In an ideal planar bilayer, there is no concept h of reduced volume, so area can be drawn from edges, unless pinned which would generate tension by area conservation What is the energy required for pulling the excess energy from the edges Can one conduct studies with pinned flat bilayer as a reference state, defining an excess area and doing a systematic analysis. The excess area will popup or down the base state of the membrane. Ve=L/Lpin Would you get or and when 80