40 years of Microdialysis

Urban Ungerstedt, Prof. emeritus

Karolinska institutet, Stockholm

Berlin, Dec 11, 2014

 Why microdialysis …

Ungerstedt U, Pycock C.
Functional correlates of dopamine neurotransmission.
Bull Schweiz Akad Med Wiss. 1974;30:44-55.

Tissue dialysis

Stereotaxic mapping of the monoamine pathways in the rat brain

Acta Physiol Scand (Supplement)1971
Brain capillary 0,01 mm ”Hollow fibre” 0,5 mm
Urban Ungerstedt 12/14/20 2
 15 513 publications

2014

1974

Microdialysis
Ginza pyramid
publications
(Egypt)
(PubMed)
Urban Ungerstedt 12/14/20 3
Microdialysis: The simple, non-
invasive, universal biosensor. TRUE?

Dialysing
membrane

Double lumen
shaft
Welcome to the Microdialysis boot-camp!

Blood Cells
capillary

Interstitial
fluid
In VITRO recovery:
1
First step toward a new study

10% recovery

10
In VIVO recovery:
<1
A very different story

Lower recovery due to

limitations in supply
and diffusion
10
Maximizing recovery

Longer membrane

Slower flow

Higher cut off

membrane
But we can do very little
about diffusion
Relative recovery – chosing flow,
chosing membrane length

100% recovery Longer membrane

Shorter membrane
mol/L

0 0,3 1 2 Flow ml/min

Absolute recovery -
chosing a ”volume-insensitive” analysis
Max diffusion in tissue

mol
Note! You may disturb
tissue physiology

0 1 2 Flow ml/min
Chosing outflow tubing

Adsorbtion to tubing saturated

mol or
mol/L

0 Time
Determining the extracellular concentration:
Extrapolation to zero flow method

Note! You may over-

estimate the concentration
Conc.

1 2 Flow ml/min
Determining the extracellular concentration:
Near equilibrium method

5 mM 5 mM 5 mM

0 mM 0.5 mM 2.5 mM 5 mM 5 mM

Low flow
Long membrane
 Near equilibrium dialysis in muscle

5,0
4,5
4,0
3,5
Glucose mM

3,0
2,5
2,0
1,5 Glucose
1,0
0,5 1.33 µl/min 0.66 µl/min 0.33 µl/min 0.16 µl/min
0,0
0 1 2 3 4 5 6 7 8
Hours
 Near equilibrium dialysis in muscle

5,0
4,5
4,0 Lowering flow
3,5
Glucose mM

3,0
2,5
2,0
1,5 Glucose
1,0
0,5 1.33 µl/min 0.66 µl/min 0.33 µl/min 0.16 µl/min
0,0
0 1 2 3 4 5 6 7 8
Hours
 Near equilibrium dialysis in muscle

5,0
Lowering flow
4,5
4,0
3,5
Glucose mM

3,0
2,5
2,0
1,5 Glucose
1,0
0,5 1.33 µl/min 0.66 µl/min 0.33 µl/min 0.16 µl/min
0,0
0 1 2 3 4 5 6 7 8
Hours
Determining the extracellular concentration:
No-net flux method

5 mM 5 mM 5 mM

1 mM 2 mM 3 mM 4 mM 4 mM

5 mM 5 mM 5 mM

9 mM 8 mM 7 mM 6 mM 6 mM
Determining the extracellular concentration:
No net flux calculation

slope = recovery
C out - C in

C¥ extrac ellular

Cin
Determining the extracellular concentration:
Internal standard method

1 10% recovery
10 9

Assumption: Recovery is the

same in both directions
 Osmosis

Water transfer Micro-

dialysis
catheter

Large
molecules
 Osmosis Rlative recovery increases!

Water transfer Micro-

dialysis
Muscle 8ml/hr catheter
sc Fat 2ml/hr

Large
molecules
 Balancing osmostic forces

Water transfer Micro-

dialysis
catheter

Large
molecules
 Recovery vs Time

Break in time Stable recovery

100%

< 100%
recovery

Time
Break in time explained:
Air trapping
Inner dialysing skin
Outer non-dialysing skin
Pores filled with air

Air must leave

Water must enter
Dipping, Flushing decreases the
Break in time
Dipping
water
Flushing
water
Wet implantation

Dry implantation
 Maximizing recovery:
Animal compared to man

Animal Man

Flow 2ml/min 0.3 ml/min 7x

Membrane 2mm 10 to 30mm 5 to 15x
Recovery (man/animal) 35 to 105x
 Microdialysis reveals very complex physiological
events where TRENDS are as important as LEVELS

This demands new methods of statistical analysis

which we are still very uncertain about
We must be especially careful when we document,
analyze and present methods as well as data

We need to examine individual data much more

carefully than we are used to in an age of statistical
do it yourself software

Some of these concerns are reflected in the 2014

Consensus statement, authored by 37 experienced
microdialysis users
Urban Ungerstedt 12/14/20 27
 Consensus statement from the 2014
International Microdialysis Forum
Publication of microdialysis data should include the
following information (Core Data reporting):
• catheter type
• catheter location
• flow rate
• membrane length
• perfusion fluid composition
• time from implantation to monitoring
Urban Ungerstedt 12/14/20 28
PLEASE use LabPilot when examining your
data - An indespensable free (!) tool designed
for the analysis of microdialysis data

Urban Ungerstedt 12/14/20 29

And now - back to the Swedish forest

Urban Ungerstedt 12/14/20 30