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

Introduction to Data Analysis

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
F_1:3

F_2:3

F_3:3

F_4:3

D_1:3

D_2:3

D_3:3

D_4:3

Time (seconds)
1E4 8000 6000 4000 2000
F
_
1
:
3
0
-5
-10
-15
-20
-25
-30
-35
-40
-45
-50
-55
F_1:3

F_2:3

F_3:3

F_4:3

D_1:3

D_2:3

D_3:3

D_4:3

Time (seconds)
1E4 8000 6000 4000 2000
F
_
1
:
3
0
-5
-10
-15
-20
-25
-30
-35
-40
-45
-50
-55
D
_
1
:
3
1,2
1
0,8
0,6
0,4
0,2
0
Less mass
more mass
Viscous/floppy/elongated
Rigid,/compressed/flat
Qualitative Analysis, cont.
0
0.1
0.2
0.3
0.4
0.5
-20 -15 -10 -5 0

D

(
1
0
-
6
)
f (Hz)
Low affinity
High affinity
1) D-f plot
Reveals reaction fingerprints, multiple phases, time independant
Antigen covered
sensor
Binding of
antibodies
-25
-20
-15
-10
-5
0

f

(
H
z
)
Low affinity
High affinity
0 500 1000 1500 2000 2500

D
Time (s)
0.2x10
-6
Low affinity
High affinity
0
0.1
0.2
0.3
0.4
0.5
-20 -15 -10 -5 0

D

(
1
0
-
6
)
f (Hz)
Low affinity
High affinity
D-f plot - Monoclonal antibodies
Quantitative analysis the
Sauerbrey equation
F_1:3

F_2:3

F_3:3

F_4:3

D_1:3

D_2:3

D_3:3

D_4:3

Time (seconds)
1E4 8000 6000 4000 2000
F
_
1
:
3
0
-5
-10
-15
-20
-25
-30
-35
-40
-45
-50
-55
D
_
1
:
3
1,2
1
0,8
0,6
0,4
0,2
0
F_1:3

F_2:3

F_3:3

F_4:3

D_1:3

D_2:3

D_3:3

D_4:3

Time (seconds)
1E4 8000 6000 4000 2000
F
_
1
:
3
0
-5
-10
-15
-20
-25
-30
-35
-40
-45
-50
-55
D
_
1
:
3
1,2
1
0,8
0,6
0,4
0,2
0
AD>>0, Sauerbrey will underestimate the mass
AD~0, Sauerbrey will give a correct mass estimate
The Sauerbrey relation:
Am[ng*cm
-2
]=-17,7[cm
2
*ng
-1
*Hz
-1
]* f [Hz]
Sauerbrey mass

Time (seconds)
1E4 8000 6000 4000 2000
S
a
u
e
r
b
r
e
y

m
a
s
s
1000
900
800
700
600
500
400
300
200
100
0
The Sauerbrey relation
Overtones scaled by overtone number (n)
The same constant can be used for all overtones
f
n
C m =
1
overtone n
s ngcm C

=
1 2
7 , 17
Linear relationship between frequency and mass/surface area:

m
=
F3/3 (Hz)
F5/5 (Hz)
D3 (1E-6)
D5 (1E-6)
Time (min)
120 110 100 90 80 70 60 50 40 30 20 10 0
F
3
/
3

(
H
z
)
0
-2
-4
-6
-8
-10
-12
-14
-16
-18
-20
-22
-24
-26
-28
-30
D
3

(
1
E
-
6
)
4
3
2
1
0
Film thickness
Qualitative analysis the
viscoelastic model
F_1:3

F_2:3

F_3:3

F_4:3

D_1:3

D_2:3

D_3:3

D_4:3

Time (seconds)
1E4 8000 6000 4000 2000
F
_
1
:
3
0
-5
-10
-15
-20
-25
-30
-35
-40
-45
-50
-55
D
_
1
:
3
1,2
1
0,8
0,6
0,4
0,2
0
AD>>0, Sauerbrey will underestimate the mass
Viscoelastic voight model
Output:
: density, (kg/m3)
: viscosity (G/), (kg/ms)
: elasticity (G), (Pa)
: thickness, (m)
Input:
f1
f3
D1
D3
Viscosity
1. Viscosity is a measure of a fluid's
resistance to flow
Newtons definition , coefficient of viscosity, viscosity
or dynamic viscosity
Unit
Pas, (which is identical
to 1 Ns/m
2
or 1 kg/ms).
y
u

=
Time
D
e
f
o
r
m
a
t
i
o
n
F
o
r
c
e
Shear modulus (Elasticity)
1. Elasticity
2. (Physics) The ratio of shearing stress to shearing strain within the
proportional limit of a material.
Unit (Pa, or N/m
2
)

= G
Time
D
e
f
o
r
m
a
t
i
o
n
F
o
r
c
e
Viscoelasticity
A viscoelastic material is, as the name
suggests, one which shows a
combination of viscous and elastic
effects.
Elastic (spring)
Viscous (dashpot)
Voight element
Viscoelastic model
f=f
1
(n,
f
,
f
,
f
,
f
)
D=f
2
(n,
f
,
f
,
f
,
f
)
Crystal
Adlayer
(
f
,
f
,
f
)
d
f
Fluid
(
l
,
l
)
n=1
n=3
n=...
Voinova et al., Physica Scripta 59 (1999) 391
G* = G' + jG''
= + j2f
: density, (kg/m3)
: viscosity (G/), (kg/ms)
: elasticity (G), (Pa)
: thickness, (m)
Introduction to fitting
Model
converged,
results given
User output
QTools
Fitting routine SIMPLEX
Nelder, J. A., & Mead, R. 1965,Comp. J., 7, 308
Compare
meas. & fun.
Calculation of
Function value
Generate new
parameters
Initial estimate of
parameters
User input
Operating range
10
0
10
1
10
2
10
3
10
4
10
5
10
6
10
7
10
8
Lab viscometers
QCM-D
Modeled output based on a narrow frequency window
Data from lower frequency range cannot necessarily be compared with QCM-
D modeled data.
Hz
Quartz crystal
Lipid film
Lipase solution
~100 nm
A practical modeling example
Lipase (E.C. 3.1.1.3)
Molecular Weight ~30kDa
Concentration 1 g/ml
Lipoprime (lipase)
Formula: C
57
H
104
O
6
Molecular Weight: 885.43 Da
CAS Registry Number: 122-32-7
Triolein (triacylglycerol)
Snabe and Petersen, Aalborg University
Chemistry and Physics of Lipids 125(2003), 69-82
Enzymatic degradation of lipid films
Raw data indicates multiphase process
Viscoelastic modeling gives additional information
Snabe and Petersen, Aalborg University
Chemistry and Physics of Lipids 125(2003), 69-82
F1 (Hz) - 5MHz F3/3 (Hz) -15MHz F5/5 (Hz) - 25MHz
Time (min)
30 20 10 0
F
r
e
q
u
e
n
c
y
,

(
H
z
)
250
200
150
100
50
0
D1 (1E-6) D3 (1E-6) D5 (1E-6)
Time (min)
30 25 20 15 10 5 0
D
i
s
s
i
p
a
t
i
o
n

(
1
e
-
6
)
40
30
20
10
0
-10
-20
-30
0
1
2
3
4
5
6
0 1 2 Time (min)
V
i
s
c

(
k
g

m
-
1

s
-
1
)

o
r

E
l
a
s
t
i
c
i
t
y

(
1
0
5

P
a
)
0
20
40
60
80
100
120
F
i
l
m

T
h
i
c
k
n
e
s
s

(
n
m
)
Quartz Crystal
Lipid film
A
A
A) Adsorption of lipase
Quartz crystal
Lipid film
B
B
B) Cluster formation
Quartz Crystal
Lipid film
C
C
C) Mass ejection
Quartz Crystal
Lipid film
D
D
D) Lipid layer removal
Enzymatic degradation of lipids
0
1
2
3
4
5
6
0 5 10 15 20
Time (min)
V
i
s
c

(
k
g

m
-
1

s
-
1
)

o
r

E
l
a
s
t
i
c
i
t
y

(
1
0
5

P
a
)
0
20
40
60
80
100
120
F
i
l
m

T
h
i
c
k
n
e
s
s

(
n
m
)
A
D
B
C
0
1
2
3
4
5
6
0 5 10 15 20
Time (min)
V
i
s
c

(
k
g

m
-
1

s
-
1
)

o
r

E
l
a
s
t
i
c
i
t
y

(
1
0
5

P
a
)
0
20
40
60
80
100
120
F
i
l
m

T
h
i
c
k
n
e
s
s

(
n
m
)
A
D
B
C
Snabe and Petersen, Aalborg University
Chemistry and Physics of Lipids 125(2003), 69-82
Thought process
Are there high
values in my
data?
Sauerbrey
D/f plot
Raw data plot
D/f plot
Raw data plot
Are the results
within the model
assumptions
Viscoelastic model
D/f plot
Raw data plot
Homogenous adlayer
Newtonian fluid
0 > D
D
If Sauerbrey
will under estimate the
thickness
Raw data, Qsoft data
file
Yes
No
Yes
No
Comments
Evaluation methods
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) Response parameter;
- frequency
- Dissipation
- Modeled thickness
) 1 ( ) (
) ( t k C k
eq
off on
e R t R
+
=
) 1 ( ) (
) (
1
t C k
eq
e R t R

=
2) Perform adsorption at different C
3) Equation system for k
1
with C and R
eq
Extract k
1
t k
eq
off
e R t R

= ) (
4) Determine k
oII
from dissociation phase
F3/3 (Hz)
D3 (1E-6)
Testdata kinetic2wfi t: 2003-09-30 15:33:00
Time (min)
6000 5500 5000 4500 4000 3500 3000 2500 2000 1500 1000 500 0
F
3
/3
(H
z
)
0
-1
-2
-3
-4
-5
-6
-7
-8
-9
-10
-11
-12
-13
-14
D
3
(1
E
-6
)
2
1
0
B+S BS
k
on
k
off
[ ]
[ ][ ]
off
on
a
k
k
S B
BS
K = =
5) Calculate k
on
from k
oII
and k
1
6) Calculate K
D
Swelling of cellulose
Cellulose coated crystal, (100nm)
EtOH H
2
O
Swelling
Susanna Flt, Mitthgskolan, Sundsvall, Sweden
High charge, more swelling
Swelling kinetics
-2000
-1500
-1000
-500
0
500
-5 0 5 10 15 20 25 30 35 40 45 50
Time (hrs)
F
(
1
5
)

H
z
20 ueq/g
409 ueq/g
-1600
-1400
-1200
-1000
-800
-600
-400
-200
0
200
0 10 20 30 40
Time (min)
F
(
1
5
)
H
z
20 ueq/g
409 ueq/g
Swelling of Cellulose
Offset e A t F
Ae y t F
k t
b t
+ =
+ =

) 1 ( ) (
) (
) *
/
0
Determination of the decay constant
F(t)= frequency
t= time
Y0=A+Offset= F at t=very large
Offset= F at t=0
b=1/k, decay constant (swelling parameter)
C
Fit C
Time (s)
1,5E5 1E5 5E4 0
F
2

(
H
z
)

[
3

*

H
z
]
0
-50
-100
-150
-200
-250
-300
-350
-400
-450
-500
-550
-600
-650
-700
-750
-800
-850
-900
-950
-1000
-1050
b ~2000
Summary
1) Qualitative, Raw data, D-f
2) Quantitative Sauerbrey
3) Quantitative Viscoelastic
4) Curve fit
F3/3 (Hz)
D3 (1E-6)
Time (min)
9 8 7 6 5 4 3 2 1 0
F
3
/3
(H
z
)
0
-2
-4
-6
-8
-10
-12
-14
-16
-18
-20
-22
-24
-26
-28
-30
-32
D
3
(1
E
-6
)
2
1
0
D3 (1E-6)
F3/3 (Hz)
0 -5 -10 -15 -20 -25 -30
D
3
(1
E
-6
)
2
1
0
F3/3 (Hz)
F5/5 (Hz)
fir f3
fit f5
D3 (1E-6)
D5 (1E-6)
fit d3
fit d5
Time (min)
8 6 4 2 0
F
3
/3
(H
z)
0
-2
-4
-6
-8
-10
-12
-14
-16
-18
-20
-22
-24
-26
-28
-30
-32
-34
D
3
(1
E
-6
)
2
1
0
Sauerbrey mass
Time (seconds)
1E4 8000 6000 4000 2000
S
a
u
e
rb
re
y
m
a
s
s
1000
900
800
700
600
500
400
300
200
100
0
C
Fit C
Time (s)
1,5E5 1E5 5E4 0
F2 (H
z) [3 * H
z]
0
-50
-100
-150
-200
-250
-300
-350
-400
-450
-500
-550
-600
-650
-700
-750
-800
-850
-900
-950
-1000
-1050
Thank you for your attention!