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VISIT IN TUBINGEN
The retinal preparation process1. Removing the eye cup of rat
2. Cut the eye cap along the ora serrata
3. Dissect the retina for three segments that will be recorded
4. Loosen the neural retina from the sclera and transfer it to the recording field of the MEA
The retinal preparation process
The perforated MEA
Improve retina - electrodes contact.
Measure the ganglion cell activity.
The perforated MEA setup
The improved contact achieved by applying a negative pressure fro underneath the MEA
MCS system
Amplifier and data acquisition.
MEAs.Light sources to
stimulate the retina.Perfusion setup.
STIMULUS OF UNIFORMLIGHT
Light intensity of 3.6klux
Cycle time
•Total time of 30 sec
•TP=time of pulse
•f= frequency.
The f and TP values
5ms50ms100ms500ms
0.25Hz
0.5Hz
1Hz
5Hz
10Hz
20Hz
The signal processing block diagram
FFT TRANSFORMATION
ROW ELECTRODE FILTER
200Hz-3kHzreconstruction
Frequency
space
Frequency
space
(peak detector)
Level filter
50mv
Time space
Process the date )frequency, number of AP, width of the
respond …(
Compare between 50,100,500 ms pulse at 0.5Hz from one electrode number 47
0 1 2 3 4 5 6 7 8 9 10-500
0
500
time [sec]
volta
getime space
0 1 2 3 4 5 6 7 8 9 10-500
0
500
time [sec]
volta
ge
time space
0 1 2 3 4 5 6 7 8 9 10-200
0
200
time [sec]
volta
ge
Frequency space
-5000 -4000 -3000 -2000 -1000 0 1000 2000 3000 4000 50000
5
10x 10
6m
ag
-5000 -4000 -3000 -2000 -1000 0 1000 2000 3000 4000 50000
2
4x 10
6
mag
-5000 -4000 -3000 -2000 -1000 0 1000 2000 3000 4000 50000
1
2x 10
6
mag
freq [Hz]
-100 0 100 200 300 400 500 600 700 800 9000
5
10x 10
6m
ag
-100 0 100 200 300 400 500 600 700 800 9000
2
4x 10
6
mag
-100 0 100 200 300 400 500 600 700 800 9000
1
2x 10
6
mag
freq [Hz]
Frequency space zoom in
Frequency space filtered
-3000 -2000 -1000 0 1000 2000 30000
5
10x 10
4
-3000 -2000 -1000 0 1000 2000 30000
5
10x 10
4
-3000 -2000 -1000 0 1000 2000 30000
5
10x 10
4
freq [Hz]
Reconstructed signal 50,100,500 mswith peak level at 50mV
0 1 2 3 4 5 6 7 8 9 10-200
0
200
time [sec]
volta
ge
0 1 2 3 4 5 6 7 8 9 10-200
0
200
time [sec]
volta
ge
0 1 2 3 4 5 6 7 8 9 10-100
0
100
time [sec]
volta
ge
50ms action potential counter
0 1 2 3 4 5 6 7 8 9 10-150
-100
-50
0
50
100
150
200
250
time [sec]
volta
ge
100ms action potential counter
0 1 2 3 4 5 6 7 8 9 10-150
-100
-50
0
50
100
150
200
250
time [sec]
volta
ge
500ms action potential counter
0 1 2 3 4 5 6 7 8 9 10-100
-50
0
50
100
150
200
250
time [sec]
volta
ge
50ms Action potential frequency
0 1 2 3 4 5 6 7 8 9 10-100
-50
0
50
100
150
time [sec]
freq
uenc
y[A
P/s
ec]
100ms Action potential frequency
0 1 2 3 4 5 6 7 8 9 10-100
-50
0
50
100
150
time [sec]
freq
uenc
y[A
P/s
ec]
500ms Action potential frequency
0 1 2 3 4 5 6 7 8 9 10-100
-50
0
50
100
150
time [sec]
freq
uenc
y[A
P/s
ec]
0 1 2 3 4 5 6 7 8 9 10-500
0
500
time [sec]
volta
getime space
0 1 2 3 4 5 6 7 8 9 10-500
0
500
time [sec]
volta
ge
time space
0 1 2 3 4 5 6 7 8 9 10-200
0
200
time [sec]
volta
ge
Compare between 50,100,500 ms pulse at 0.25Hz from one electrode number 47
-5000 -4000 -3000 -2000 -1000 0 1000 2000 3000 4000 50000
2
4x 10
6m
ag
-5000 -4000 -3000 -2000 -1000 0 1000 2000 3000 4000 50000
1
2x 10
6
mag
-5000 -4000 -3000 -2000 -1000 0 1000 2000 3000 4000 50000
1
2x 10
6
mag
freq [Hz]
Frequency space
-100 0 100 200 300 400 500 600 700 800 9000
2
4x 10
6m
ag
-100 0 100 200 300 400 500 600 700 800 9000
1
2x 10
6
mag
-100 0 100 200 300 400 500 600 700 800 9000
1
2x 10
6
mag
freq [Hz]
Frequency space zoom in
-3000 -2000 -1000 0 1000 2000 30000
5
10x 10
4
-3000 -2000 -1000 0 1000 2000 30000
5
10x 10
4
-3000 -2000 -1000 0 1000 2000 30000
5
10x 10
4
freq [Hz]
Frequency space filtered
0 1 2 3 4 5 6 7 8 9 10-200
0
200
time [sec]
volta
ge
0 1 2 3 4 5 6 7 8 9 10-200
0
200
time [sec]
volta
ge
0 1 2 3 4 5 6 7 8 9 10-200
0
200
time [sec]
volta
ge
Reconstructed signal 50,100,500 mswith peak level at 50mV
0 1 2 3 4 5 6 7 8 9 10-150
-100
-50
0
50
100
150
200
250
time [sec]
volta
ge50ms action potential counter
0 1 2 3 4 5 6 7 8 9 10-150
-100
-50
0
50
100
150
200
250
time [sec]
volta
ge100ms action potential counter
0 1 2 3 4 5 6 7 8 9 10-200
-150
-100
-50
0
50
100
150
200
250
time [sec]
volta
ge500ms action potential counter
0 1 2 3 4 5 6 7 8 9 10-200
-150
-100
-50
0
50
100
150
time [sec]
freq
uenc
y[A
P/s
ec]
50ms Action potential frequency
0 1 2 3 4 5 6 7 8 9 10-200
-150
-100
-50
0
50
100
150
200
time [sec]
freq
uenc
y[A
P/s
ec]
100ms Action potential frequency
0 1 2 3 4 5 6 7 8 9 10-200
-150
-100
-50
0
50
100
150
200
time [sec]
freq
uenc
y[A
P/s
ec]
500ms Action potential frequency
The total result of all the electrodestimulus of 0.5Hz 50ms
0 5 10 15 20 25 30 350
1000
2000
3000
4000
5000
6000
7000Total number of action potential
time[sec]
Tot
al n
umbe
r of
act
ion
pote
ntia
l
Number of action potential at 10ms stimulus of 0.5Hz 50ms
0 5 10 15 20 25 30 350
2
4
6
8
10
12
14
16
18
20number of AP)time(
time[sec]
num
ber
of a
ctio
n po
tent
ial a
t 10
ms
Action potential frequency )calculated at
100ms( stimulus of 0.5Hz 50ms
0 5 10 15 20 25 30 350
200
400
600
800
1000
1200action potential frequency
time [sec]
freq
uenc
y[A
P/s
ec]
The total result of all the electrodestimulus of 0. 5Hz 100ms
0 5 10 15 20 25 30 350
1000
2000
3000
4000
5000
6000Total number of action potential
time[sec]
Tot
al n
umbe
r of
act
ion
pote
ntia
l
Number of action potential at 10ms stimulus of 0.5Hz 100ms
0 5 10 15 20 25 30 350
2
4
6
8
10
12
14
16
18
20number of AP/10ms)time(
time[sec]
num
ber
of a
ctio
n po
tent
ial a
t 10
ms
Action potential frequency )calculated at
100ms( stimulus of 0.5Hz 100ms
0 5 10 15 20 25 30 350
100
200
300
400
500
600
700
800
900
1000action potential frequency
time [sec]
freq
uenc
y[A
P/s
ec]
The total result of all the electrodestimulus of 0.5Hz 500ms
0 5 10 15 20 25 30 350
500
1000
1500
2000
2500
3000
3500
4000
4500Total number of action potential
time[sec]
Tot
al n
umbe
r of
act
ion
pote
ntia
l
Number of action potential at 10ms stimulus of 0.5Hz 500ms
0 5 10 15 20 25 30 350
2
4
6
8
10
12number of AP/10ms)time(
time[sec]
num
ber
of a
ctio
n po
tent
ial a
t 10
ms
Action potential frequency )calculated at 100ms( stimulus of 0.5Hz 500ms
0 5 10 15 20 25 30 350
100
200
300
400
500
600
700
800action potential frequency
time [sec]
freq
uenc
y[A
P/s
ec]
Zoom in at the stimulus of 0.5Hz 500 ms pulse counter.
0 1 2 3 4 5 6 70
500
1000
1500
2000
2500Total number of action potential
time[sec]
Tot
al n
umbe
r of
act
ion
pote
ntia
l
Zoom in at the stimulus of 0.5Hz 500 ms )number of pulse at 10ms (
0 1 2 3 4 5 6 70
2
4
6
8
10
12number of AP/10ms)time(
time[sec]
num
ber
of a
ctio
n po
tent
ial a
t 10
ms
0 1 2 3 4 5 6 70
100
200
300
400
500
600
700
800action potential frequency
time [sec]
freq
uenc
y[A
P/s
ec]
Zoom in at the stimulus of 0.5Hz 500 ms )frequency (
Experiment in chicken retina
Focusing image from computer display on the retina.
the space and time frequency can be controlled.
the stimulus is from the bottom
The optic setup
45 d mirror. Lance to focus the
image. Mechanical setup. display that radiate
the image.
Movie of the experiment
The buffer components ratchicken
mMg/molg/1litermMg/molg/1liter
NaCl12058.447.01281005.84
KCl574.560.372860.4476
NaHCO33084.012.5203302.52
GLUKOSE15180.162.7024509.01
L-GLUTAMAT0.2169.10.03382
MgCl2 6H2O1203.30.203320.493
PHENOLROT1ml
ph7.57.5
OSMOLARIT333326-328
NaH2PO4)1H2O(10.138
CaCl2)2H20(10.147
The retina preparation
