Introduction to Data Analysis

Q-Sense Basic Training, April 4-5, 2006

Outline
Different types of data evaluation
Functions in QTools
Introduction to viscoelastic modeling

Analysis Methods
1) Qualitative analysis (raw data plot, D-f plot)
2) Quantitative analysis (low D=Sauerbrey)
3) Quantitative analysis (high D=viscoelastic
modeling)
4) Curve fit functions

Qualitative Analysis
1) Raw data plot, relative comparison of responses
Viscous/floppy/elongated

F_1:3

more mass

D_1:3

F_1:3

Less mass

Time (seconds)
F_1:3

F_2:3

F_3:3

F_4:3

D_1:3

D_2:3

D_3:3

Time (seconds)

D_4:3

F_1:3

F_2:3

F_3:3

F_4:3

D_1:3

D_2:3

D_3:3

D_4:3

Rigid,/compressed/flat

Qualitative Analysis, cont.

1) D-f plot
0.5
Low affinity

0.3

-6

D (10 )

0.4

0.2

High affinity

0.1
0
0

-5

-10
f (Hz)

-15

-20

Reveals reaction fingerprints, multiple phases, time independant

D-f plot - Monoclonal antibodies

0.5
0.4

High affinity

-15

0.3
0.2

High affinity

0.1
0

-20
0

-25

-5

High affinity

Low affinity

Antigen covered
sensor

f (Hz)

-10

Low affinity

-6

Low affinity

-5

D (10 )

0.2x10-6

500

1000 1500 2000 2500

Time (s)

Binding of
antibodies

-10
f (Hz)

-15

-20

Quantitative analysis the

Sauerbrey equation
D>>0, Sauerbrey will underestimate the mass

The Sauerbrey relation:

m[ng*cm-2]=-17,7[cm2*ng-1*Hz-1]* f [Hz]

Sauerbrey mass

F_1:3

D_1:3

Time (seconds)
F_2:3

F_3:3

F_4:3

D_1:3

D_2:3

D_3:3

D_4:3

D~0, Sauerbrey will give a correct mass estimate

Time (seconds)
F_1:3

F_2:3

F_3:3

F_4:3

D_1:3

D_2:3

D_3:3

D_4:3

D_1:3

F_1:3

F_1:3

Time (seconds)
Sauerbrey mass

The Sauerbrey relation

Linear relationship between frequency and mass/surface area:

1
m = C f
n

C = 17,7ngcm 2 s 1
n overtone

Film thickness

F3/3 (Hz)

D3 (1E-6)

F3/3 (Hz)
F5/5 (Hz)
D3 (1E-6)
D5 (1E-6)

Time (min)

Overtones scaled by overtone number (n)

The same constant can be used for all overtones

Qualitative analysis the

viscoelastic model
D>>0, Sauerbrey will underestimate the mass

F_1:3

D_1:3

Time (seconds)
F_1:3

F_2:3

F_3:3

F_4:3

D_1:3

D_2:3

D_3:3

Input:

f1
f3
D1
D3

Viscoelastic voight model

D_4:3

Output:
: density, (kg/m3)
: viscosity (G/ ), (kg/ms)
: elasticity (G), (Pa)
: thickness, (m)

Viscosity

Force

Deformation

1. Viscosity is a measure of a fluid's

resistance to flow

Newtons definition

, coefficient of viscosity, viscosity

or dynamic viscosity
=

Time

u
y

Unit
Pas, (which is identical
to 1 Ns/m2 or 1 kg/ms).

Shear modulus (Elasticity)

Elasticity
(Physics) The ratio of shearing stress
proportional limit of a material.

to shearing strain

within the

Force

Deformation

1.
2.

G =

Time

Unit (Pa, or N/m2)

Viscoelasticity
A viscoelastic material is, as the name
suggests, one which shows a
combination of viscous and elastic
effects.
Voight element
Viscous (dashpot)

Elastic (spring)

Viscoelastic model
f=f1(n, f, f, f, f)
D=f2(n, f, f, f, f)

Fluid

( l, l)

: density, (kg/m3)
: viscosity (G/ ), (kg/ms)
: elasticity (G), (Pa)

n=...
n=1
n=3

Adlayer

( f, f, f)

: thickness, (m)

Crystal

G* = G'+ jG'
'
= + j2 f
Voinova et al., Physica Scripta 59 (1999) 391

df

Introduction to fitting
Initial estimate of
parameters

Calculation of
Function value

Compare
meas. & fun.

Model
converged,
results given

User input

QTools

Generate new
parameters

User output

Fitting routine SIMPLEX

Nelder, J. A., & Mead, R. 1965,Comp. J., 7, 308

Operating range
Lab viscometers

QCM-D

Hz
100

101 102 103 104 105 106 107

108

Modeled output based on a narrow frequency window

Data from lower frequency range cannot necessarily be compared with QCMD modeled data.

A practical modeling example

Lipoprime (lipase)
Lipase solution

Lipase (E.C. 3.1.1.3)

Molecular Weight ~30kDa
Concentration 1 g/ml

Lipid film

Triolein (triacylglycerol)

~100 nm

Quartz crystal

Formula: C H O
Molecular Weight: 885.43 Da
CAS Registry Number: 122-32-7

Snabe and Petersen, Aalborg University

Chemistry and Physics of Lipids 125(2003), 69-82

Frequency, (Hz)

Dissipation (1e-6)

Enzymatic degradation of lipid films

Time (min)

Time (min)
F1 (Hz) - 5MHz

F3/3 (Hz) -15MHz

F5/5 (Hz) - 25MHz

D1 (1E-6)

D3 (1E-6)

D5 (1E-6)

Raw data indicates multiphase process

Viscoelastic modeling gives additional information

Snabe and Petersen, Aalborg University

Chemistry and Physics of Lipids 125(2003), 69-82

Enzymatic degradation of lipids

5
4

120

100

80

60

40

100
80

20

120

0
0

Time (min) 1

60

40

20

A) Adsorption of lipase
B) Cluster formation
0
C) Mass ejection

D) Lipid layer removal

Film Thickness (nm )

Visc (kg m-1 s-1) or Elasticity (105

Pa)

6
Visc (kg m-1 s-1 ) or Elasticity (105
Pa)

A B

Film Thickness (nm)

C
B
A

0
5

Lipid10
film

A
C
B
D

15

20

Time
(min)
Quartz
crystal
Crystal
Snabe and Petersen, Aalborg University
Chemistry and Physics of Lipids 125(2003), 69-82

Thought process
Comments
If

D > 0

Sauerbrey
will under estimate the
thickness

Raw data, Qsoft data

file

Are there highD

values in my
data?

Evaluation methods
No

Sauerbrey
D/f plot
Raw data plot

No

D/f plot
Raw data plot

Yes

Homogenous adlayer
Newtonian fluid

Are the results

within the model
assumptions

Yes

Viscoelastic model
D/f plot
Raw data plot

Curve fitting functions

Fitting of of f and D data to
1) Predefined adsorption models
2) User defined equations

Method: Determination of kinetic

constants with QCM-D
1)

2)

Response parameter;
- frequency
- Dissipation
- Modeled thickness

4)

Determine k

from dissociation phase

R(t ) = Req e

k off t

Perform adsorption at different C

F3/3 (Hz)
D3 (1E-6)

F3/3 (Hz)

Ka =

[BS ] = kon
[B][S ] koff

D3 (1E-6)

R (t ) = Req (1 e

( k on C + k off ) t

B+S

R (t ) = Req (1 e
3)

( k1C ) t

Time (min)
Testdata kinetic2wfit: 2003-09-30 15:33:00

5)

Calculate k from k

6)

Calculate K

Equation system for k with C and R

kon
koff

BS

Swelling of cellulose
Cellulose coated crystal, (100nm)
500
200

High charge, more swelling

0
-200
-400

F (15) Hz

-600

20 ueq/g
409 ueq/g

-800
-1000

Swelling kinetics

-1200
-1400

-500

-1600

F(15) Hz

10

20

30

40

Time (min)

20 ueq/g
409 ueq/g

-1000

-1500

-2000
-5

10

15

20

25

30

35

40

45

50

Time (hrs)

EtOH

Swelling

H2O

Susanna Flt, Mitthgskolan, Sundsvall, Sweden

Swelling of Cellulose
Determination of the decay constant
C
Fit C

F (t ) = A(1 e t*k ) ) + Offset

F(t)= frequency

F2 (Hz) [3 * Hz]

F (t ) = y0 + Ae t / b

t= time
Y0=A+Offset= F at t=very large
Offset= F at t=0
b=1/k, decay constant (swelling parameter)

Time (s)

b ~2000

Summary
D3 (1E-6)

F3/3 (Hz)

D3 (1E-6)

1) Qualitative, Raw data, D-f

D3 (1E-6)

F3/3 (Hz)
D3 (1E-6)

F3/3 (Hz)

2) Quantitative Sauerbrey

Sauerbrey mass

Time (min)

Time (seconds)
Sauerbrey mas s

F3/3 (Hz)

D3 (1E-6)

3) Quantitative Viscoelastic

F3/3 (Hz)
F5/5 (Hz)
fir f3
fit f5
D3 (1E-6)
D5 (1E-6)
fit d3
fit d5

Time (min)

4) Curve fit

F2 (Hz) [3 * Hz]

C
Fit C

Time (s)

Thank you for your attention!