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ME/CHE/BME 6782

Cellular Engineering

Notes for 11/2 Lecture

Measuring Shear Modulus


of RBC Membrane
Micropipette Aspiration
and Shear Flow
Scanning Electron Micrograph
of a Human Red Blood Cell
The Red Blood Cell Membrane
Approaches for Measuring
Mechanical Properties of Cells
Apply known force, measure deformation
Indentation experiment
Micropipette aspiration
Flow chamber
Magnetic twisting cytometry

Take advantage of random forces and Brownian motions


Red blood cell membrane flickering
Thermal fluctuations of actin filaments and microtubules
Approaches for Measuring
Mechanical Properties of Cells
Apply known force, measure deformation
Indentation experiment
Micropipette aspiration
Flow chamber
Magnetic twisting cytometry

Take advantage of random forces and Brownian motions


Red blood cell membrane flickering
Thermal fluctuations of actin filaments and microtubules
Approaches for Measuring
Mechanical Properties of Cells
Apply known force, measure deformation
Indentation experiment
Micropipette aspiration
Flow chamber
Magnetic twisting cytometry

Take advantage of random forces and Brownian motions


Red blood cell membrane flickering
Thermal fluctuations of actin filaments and microtubules
Measuring Shear Modulus
of RBC Membrane
Difficulties
Small size and complex geometry
Non-uniform and time dependent stress/strain
Reversed Problem Approach
Assume a stress-strain relationship with unknown coefficients
Solve the boundary value problem
Fit the solution to the data to test the validity of the model
Start over with a new model if the test result is negative
Evaluate the parameter values if the test result is positive
Problem
Parameter values may be model- and technique-dependent
Solution
Conduct separate experiments using distinct techniques
Mechanical Properties of
Human Red Blood Cells
Mechanical model for the human red blood cell
A liquid bag enclosed by a membrane
The liquid can be modeled as a Newtonian viscous fluid
The membrane can be modeled as an elastic sheet

Mechanical properties of human red blood cells


Membrane shear modulus: = 6-9 pN/m = 6-9 10-6 N/m
Membrane area modulus: K = 4.5 105 pN/m = 0.45 N/m
Membrane bending modulus: B = 44 5 kBT
Hemoglobin solution viscosity : = 6 10-3 N-s/m2
Force balance (Law of LaPlace)
Micropipette p0 p = 2Tm/Rp
Aspiration
Neglecting friction at the pipette wall
Experiment
T = Tm

Free body diagram


T RBC membrane
Model as an
Neglect infinite sheet
constraint of elastic
membrane
of the p0
Tm r
2Rp p
RBC
Closed
Polar coordinates
shape
T
Stress Analysis of an Elementary Membrane Segment
Tr + dTr dr Re-arranging terms:
d/2 d/2 - dTr = (Tr - T)dr/r
Axisymmetric Integrating:
Problem T + dT rd T - Tr|r + Tr|r = Rp
Tr
= Rp (Tr - T)dr/r

Tr and T are 0
functions of r r LaPlace law:
only Tr|r = Rp = pRp/2
d
Re-arranging terms:
p = (2/Rp) Rp (Tr - T)dr/r

Force balance in the radial direction:


(Tr + dTr)(r + dr)d - Trrd sin(d/2)Tdr - sin(d/2)(T + dT)dr = 0
rTr/r + Tr - T = 0
Force balance in the circumferential direction:
cos(d/2)(T + dT)dr - cos(d/2)Tdr = 0
T/ = 0
Analysis of Deformation
r0
Area prior to Hemi-spherical cap:
deformation r
1 = 2Rp2
0 = r02 Cylindrical segment:
mapping 2 = 2Rp(L Rp)
Area remains Circular ring:
r0
constant 3 = (r2 Rp2)
r
during
deformation Total area:
0 = = 1 + 2 + 3

Area modulus K = 4.5 105 pN/m >> shear modulus = 6-9 pN/m
Constant area: r02 = 2Rp2 + 2Rp(L Rp) + (r2 Rp2)
Radial stretch ratio: r2 r02/r2 = 1 + (Rp2/r2)(2L/Rp - 1)
Differentiation: dr2/(1 - r2) = 2dr/r
Boundary conditions: r2|r = 1 r2|r = Rp = 2L/Rp
Stress-Strain Relation
r, r

Normal (r )/2 (r2 2)/2


Stresses or
, ,
Strains or
( r)/2 (2 r2)/2
Stretch ratios

r, r Shear stresses or strains

Area strains: r 1 Constant area: r = 1


Assume linear elasticity:
Tr - T = (r2 2) = (r2 r-2)
Re-arranging terms:
Tr = (r2 2)/2 T = (2 r2)/2
Solution

p = (2/Rp) Rp (Tr - T)dr/r

= (2/Rp) Rp (r2 r-2)dr/r
1 p
= (/Rp) 2l/Rp (r2 r-2)dr2/(1 - r2)
Slope = 2/Rp
1
= - (/Rp) 2l/Rp (1 + r-2)dr2
= (/Rp)[2L/Rp - 1 + ln(2L/Rp)] L/Rp

Is the above solution applicable for L < Rp?


Why or why not?
Measuring the BRC Shear Modulus
by the Flow Chamber
dx
Equilibrium:
2d(Txy) = dA
v 2yTx + Integration:
2y 2yTx
d(2yTx) T = A/y
x 0

0dA

dA0 dA

y0 y
0
x0 dx0 x dx
mapping
(x0,y0) (x,y)
Analysis of Deformation
x0 = R0 (1 - cos0) Asector = R020
R0
y0 = R0sin0 Atriangle = R0sin0 R0cos0
0
A0 A0 = R02(0 - sin0 cos0)
cos0 = 1 - x0/R0
0 = cos-1(1 - x0/R0) Constant area:
R0cos0
sin0 = [1 (1 - x0/R0)2]1/2 A = A0 = R02{cos-1(1 - x0/R0)
(1 - x0/R0)[1 (1 - x0/R0)2]1/2}
Stretch ratios: x = dx/dx0 y = y/y0
Constant area: xy = 1 y = x-1
Expressing y as a function of x0:
y = y0y = R0sin0/x = R0sin[cos-1(1 - x0/R0)](dx0/dx)
Stress-strain relation: Tx = (/2)(x2 x-2)
Re-arranging: (/2)[(dx/dx0)2 (dx/dx0)-2] = 0A/y
= 0R02{cos-1(1 - x0/R0) (1 - x0/R0)[1 (1 - x0/R0)2]1/2}/{R0sin[cos-1(1 - x0/R0)]}(dx/dx0)
(dx/dx0)4 (0R0/)(dx/dx0)3{cos-1(1 - x0/R0)[1 (1 - x0/R0)2]-1/2 - (1 - x0/R0)} = 1
Comparison to Experiment
Calculated shapes resemble those observed

10 m 10 m
Comparison to Experiment
The predicted 0R0/ vs. L/L0 curve is nearly linear

0R0
Slope = Stheor
Slope = Sexp

L/L0

= Cexp/Ctheor
Summary of Steps

Equilibrium force balance, stress analysis


Deformation mapping from the undeformed to the
deformed shape, analysis of strains or stretch ratios
Stress-strain relation constant area, linear elasticity
Solving the math problem
Comparison to experiment
Red Blood Cell Membrane Undulations