|
Neurofilament-specific antibody stain
of spinal cord neuron in culture.
(Lynn Gargan, March, 1996)
Experimental System: |
Murine monolayer networks in culture growing
on thin film electrode arrays. |
Array Plates: |
5 x 5 cm x 1.2 mm soda lime barrier
glass (1,200Å layer of SiO2) 64 ITO thin film microelectrodes terminating
in a 0.8 x 0.8 mm recording area. |
Insulation: |
Polysiloxane (DC 648)
(hydrophobic) |
Deinsulation: |
Single laser shots, pulsed nitrogen laser
(337 nm, 2 µJ/µm2) |
Tissue Source: |
Mouse Embryo CNS tissue (14 days gestation) dissociated
and seeded at 500k on 600 mm2 surface areas. |
Networks : |
Randomized cell body adhesion;
Spontaneously Active;
Minimum Size: 1 mm2 (50 neurons)
Maximum: 600 mm2 (30k neurons) |
Surface Preparation: |
Oxidation of CH3 groups to OH- groups via
flaming (1 s exposure to propane or butane flame). Addition of
poly-D-lysine and addition of laminin. |
Live neuron near row of electrode craters. Indium-tin
oxide conductors are 10 µm wide and 1200 Å thick. Craters
in 2 µm thick polysiloxane insulation are generated with
single laser pulses. |
SUMMARY OF SURVIVAL TIMES ATTAINED
(as of 12/96)
NOTE: No specific
longevity efforts were made. Data represent normal maintenance
of cultures that were not selected for experiments at earlier
times. All 'old timers' showed spontaneous activity and were
pharmacologically responsive.
Tissue |
No. of Experiments |
Maximum Survival Time
in vitro |
Size Range of Somata |
spinal cord |
792 |
312 d.i.v. |
10 - 45 µm |
cortex (frontal) |
58 |
81 d.i.v. |
---- |
cortex (auditory) |
56 |
148 d.i.v |
5 - 17 µm |
olfactory bulb |
67 |
172 d.i.v. |
---- |
cerebellum |
6 |
---- |
---- |
hippocampus |
4 |
---- |
---- |
3. Multichannel
Recording Work Station
|
3. Multi-Microelectrode
Plates
(MMEPs)
|
 |
MMEP version 3B
BasePlate: |
glass
5 x 5 cm
x 1.1 mm |
Conductors: |
indium tin oxide
(ITO)
1200 Å
thick, 10 µm wide |
Pattern: |
0.8 mm x 0.8
mm recording matrix
4 rows, 16
columns
40 µm
(center to center) between columns
200 µm
between rows |
Contacts: |
32 left, 32
right
5 mm long
0.75 mm wide
spacing:
0.75 mm |
|
 |
MMEP version 4
BasePlate: |
glass
5 x 5 cm
x 1.1 mm |
Conductors: |
indium tin oxide
(ITO)
1200 Å
thick, 8.0 µm wide
24
µm x 24 µm terminal pads |
Pattern: |
equally spaced
electrodes
150 µm
center to center
1.2 mm x
1.2 mm recording matrix |
Contacts: |
32 left, 32
right
5 mm long
0.75 mm wide
spacing:
0.75 mm |
|
|
|
|
|