Tissue Optics

Steven L. Jacques
jacquess@ohsu.edu
http://omlc.ogi.edu

Depts. of Biomedical Engineering

and Dermatology
Oregon Health & Science University,
Portland OR, USA

1. Optical properties
2. How to measure
optical properties
3. Light transport
4. Complex tissues
Consider a cup of coffee with cream.
Shine a laser onto liquid.

The reflectance is R.

Now, add some water to the coffee/cream.

Does the R increase,

decrease,
or remain the same?
 n
a
a L s'
T =e =e

Step size = 1/s

Total photon path = L n/s

n steps
before
escape out
surface
 a n
a n
L f s' a
Add water... f f s'
T =e =e =e
V + V
f =
V

Step size = 1/(s/f)

n steps
before
escape out Total photon path = L n/(s/f)
surface
 L

a L
T =e
 a L
R=e

L
 a L
R=e

a L = log e ( R)

Contributions of various absorbing molecules

 a L
R e
( a + a )L
add a drop of
absorber
R2 e
a

R2
ln
R
L L=
a
 L R R 0.3 to 0.8

a L
R=e s/a
a A
e
A A
' A 7 to 8
3 s +1
a
=e s/a
 Measurement vs properties grid

Collection
fibers 1.5

1 Collection fiber

z
r2
z [cm]
r2 = 1.0 cm
z 0.5

0 Source fiber Collection fiber

r1 = 0.3 cm

r1
x -0.5
-1 -0.5 0
x [cm]
0.5 1

Source fiber
x
 diffusion length

optical penetration
depth

transmission
collected by
fiber #1

transmission
collected by
fiber #2
black x = s < 10a blue = const s', red = const a
1
10
r1 = 0.1 cm 100
r2 = 1.0 cm s '
10

0
10 1000
T2 [1/cm ]
2

1.0
0.01
-1
a 0.1
10

1.0

-2
10 0 2 4
10 10 10
T1 [1/cm2]
black x = s < 10a blue = const s', red = const a
1
10
r1 = 0.3 cm 100
r2 = 1.0 cm s '
10

0
10 1000
T2 [1/cm ]

As r1 r2
2

1.0
T1 T2 ,
0.01
-1
a 0.1 so only one measurement,
10 and the grid collapses.
One no longer can specify
1.0 a and s,
just the ratio s/a .

-2
10 0 2 4
10 10 10
T1 [1/cm2]
black x = s < 10a blue = const s', red = const a
1
10
r1 = 0.6 cm 100
r2 = 1.0 cm s '
10

0
10 1000
T2 [1/cm ]

As r1 r2
2

1.0
T1 T2 ,
0.01
-1 a0.1 so only one measurement,
10 and the grid collapses.
One no longer can specify
1.0 a and s,
just the ratio s/a .

-2
10 0 2 4
10 10 10
T1 [1/cm2]
 blue = const s', red = const a
1
10
r1 = 0.9 cm 100
r2 = 1.0 cm '
s
10

0
10 1000
T2 [1/cm ]

As r1 r2
2

1.0
T1 T2 ,
0.01
a0.1
-1
so only one measurement,
10 and the grid collapses.
One no longer can specify
1.0 a and s,
just the ratio s/a .

-2
10 0 2 4
10 10 10
T1 [1/cm2]
a = 0.1 cm-1
a = 0.1 cm-1
a = 0.1 cm-1
a = 0.1 cm-1
a = 0.1 cm-1
black x = s < 10a blue = const s', red = const a
1
10
r1 = 0.1 cm 100
r2 = 1.0 cm s '
10

0
10 1000
T2 [1/cm ]
2

1.0
0.01
-1
a 0.1
10

1.0

-2
10 0 2 4
10 10 10
T1 [1/cm2]
 Oblique R(r)
Light is delivered at an angle by a optical fiber (or laser beam). The light is launched a distance of one mfp =
1/s into the tissue at an angle . The center of the diffusion process is offset from the point of point of photon
entry by x. Hence, two measurements (x, ) ----> a, s.

CCD
CCD camera with tilted source camera
0.0
abs(1/slope)
optical
fiber
slope
row

1000.0

0.0
col

1000.0 2000.0 3000.0 4000.0

ag_labv

1
= 2 mfp'
3 a (a + 's ) a =
3
' 1
1 s = a
mfp' = mfp'
a + 's
 Oblique R(r)
Light is delivered at an angle by a optical fiber (or laser beam). The light is launched a distance of one mfp =
1/s into the tissue at an angle . The center of the diffusion process is offset from the point of point of photon
entry by x. Hence, two measurements (x, ) ----> a, s.

CCD
CCD camera with tilted source camera
0.0
abs(1/slope)
optical
fiber

slope
row

1000.0

0.0
col

1000.0 2000.0 3000.0 4000.0

ag_labv

1
= 2 mfp'
3 a (a + 's ) a =
3
' 1
1 s = a
mfp' = mfp'
a + 's
black x = s < 10a blue = const s', red = const a
1
10

Oblique R(r)
a
0 0.1 0.01 0.001
10 1.0
1.0
mfp' [cm]

-1
10 10 s '

-2
10 100

-3
10 1000
-2 -1 0 1 2
10 10 10 10 10
delta [cm]
 R(r)
Reflectance as a function of
source-detector separation
 Camera
Beam of light
Mtotal.std

Standard reflectance:
example: Rstd = 0.97

Spectralon
Teflon
thick stack of white paper
 Mtotal
Rstd
Mtotal.std R
Camera
Beam of light
Mtotal
M(r)
M(r)

Test material:
Intralipid phantom
Biological tissue
 mua = 0

mua = 0.1
 mua = 0

mua = 0.5
 mua = 0

mua = 0.5
 -1/slope
 Mtotal
Rstd
Mtotal.std R
Mtotal M(r)
M(r)

Mtotal.std
r
black x = s < 10a blue = const s', red = const a
1
10
0.01 1.0
'
s
0.1
10
a
0
10
100
1.0
delta

1000
-1
10

-2
10
0.2 0.4 0.6 0.8 1
Rtotal

R
Added absorber method

Detector senses total R

Beam of light
Mstd
Reflectance
Standard
Rstd
Added absorber method

Detector senses total R

Beam of light
M
Added absorber method

Beam of light Detector senses total R

M
Added absorber method

Detector senses total R

Beam of light
Add an absorber
Added absorber method

Detector senses total R

Beam of light
Add an absorber
Added absorber method

Detector senses total R

Beam of light
+ Add an absorber
Added absorber method

Detector senses total R

Beam of light
Add more an absorber
Added absorber method

Detector senses total R

Beam of light
+ more added an absorber
 blue = const s', red = const a
1
1000
100
0.9 a = 0.00 cm-1
10
0.8
R total + added absorber

0.7
1.0
0.6
0.01
0.5

0.4

0.3
a 0.1
0.2

0.1
1.0
0
0 0.2 0.4 0.6 0.8 1
R total
 blue = const s', red = const a
1
-1 1000
0.9 a = 0.10 cm
100
0.8
R total + added absorber

0.7
10
0.6

0.5

0.4

0.3
1.0
0.01
0.2 a 0.1
0.1
1.0
0
0 0.2 0.4 0.6 0.8 1
R total
 blue = const s', red = const a
1

0.9 a = 1.00 cm-1

1000
0.8
R total + added absorber

0.7
100
0.6 s '
0.5

0.4

0.3 10

0.2

0.1
1.0
a 0.1 1.0
0.01
0
0 0.2 0.4 0.6 0.8 1
R total
 blue = const s', red = const a
1

0.9 a = 10.00 cm-1

0.8
R total + added absorber

0.7
1000
0.6

0.5

0.4

0.3 100

0.2
s '
0.1
10
1.0 a 0.1 1.0
0.01
0
0 0.2 0.4 0.6 0.8 1
R total
 M tissue S Ttissue Gtissue D
M= =
M std S Tstd Gstd D

 M tissue S Ttissue Gtissue D

M= =
M std S Tstd Gstd D

 M tissue S Ttissue Gtissue D

M= =
M std S Tstd Gstd D

Gtissue
= Ttissue = Ttissuecalib
Tstd Gstd

 M tissue S Ttissue Gtissue D
M= =
M std S Rstd Gstd D
light beam delivered,
detector distant from surface Gtissue
= Rtissue = Rtissuecalib
Rstd Gstd

know Rstd, but dont know
optical properties of standard
 M tissue S Ttissue Gtissue D
M= =
M std S Tstd Gstd D

delivery and collection Gtissue

fibers contact surface...
= Ttissue = Ttissuecalib
Tstd Gstd

cannot know Tstd

without knowing optical properties of standard
 Non-diffusive light transport

examples:
Confocal microscopy (CM)
Optical coherence microscopy (OCT)
Reflectance-mode Confocal
Microscopy
 e z

CM

Coherence gate of OCT provides a 45-fs

time slice that rejects ~99.9% of the noise.

45 fs
time Area under
slice curve is total
diffuse
reflectance R.
 e z

CM

 e z

CM

~1/1000th

OCT
 = local reflectivity [-]
= attenuation [cm-1]

z
R(z) e

 = local reflectivity [-]
= attenuation [cm-1]

z
R(z) e
s = scattering coefficient [cm-1]
?
g = anisotropy of scattering [-]

 a(g) s z f G
Tin = e
a

g
 a(g) s z f 2G
Tin Tout = e
a

g
 b/biso
= s Lf b(g)
b

g
 z f a(g ) s 2Gz f
R = Tin Tout = e = szb(g)e
 l = 488 nm

g
mouse tissues

s
0.1 m dia,
2.5% vol. fraction
polystyrene

Mie
theory
so why bother with this pedantic
pile of purposeless preoccupation
with optical properties?

glad you asked.

Consider a rCLSM or OCT image
Consider a rCLSM or OCT image
Consider a rCLSM or OCT image

R(z) @x=1mm
 1e 1 z

E1 2e 2 z

z1

1 dz
E 2 1e 1 z
E1 = e 0

z2
2 dz
E 2 = E1e z1

R


perturbation normal
 example characterizing perturbations in
cardiovascular vessel wall

 Osteogenesis Mouse skin
imperfecta
oim = Mouse
Osteogenesis
model imperfecta mutation

Ravi
Samatham

scanning
down into
skin

with Paul Campagnola, Univ. of Conn. Medical Center

 mutant

wildtype
 In vivo optical properties of
retinal layer and choroid
Ricky
Wang

Retina
800-nm OCT image z [mm] [-]
 Optical characterization of smooth muscle cell
remodeling of collagen gels
David
Levitz

Day 1 Day 5
Smooth muscles cells cause the scattering of collagen
gels to shift to lower anisotropy (g),
as if smaller structures develop that scatter
isotropically.
 Breast cancer lymph node

Robert
McLaughlin
Univ. of
Western
Australia
 [dimensionless]

Robert
McLaughlin
Univ. of
Western
Australia
 [cm-1]

Robert
McLaughlin
Univ. of
Western
Australia
 Tissue Optics

Steven L. Jacques
jacquess@ohsu.edu
http://omlc.ogi.edu

Depts. of Biomedical Engineering

and Dermatology
Oregon Health & Science University,
Portland OR, USA

1. Optical properties
2. How to measure optical
properties
3. Light transport
4. Complex tissues