3.1.3 Wet Etch and
Release
1. Remove all of the electroplating mold with PRS 3000 stripper
and rinse off again.
2. Remove the gold seed layer and adhesion promotion layer with
the wet etcher, after that the parallel electrodes are patterned.
The interdigital electrodes are 100 μm wide and 500 μm spaced
(see Note 16).
3. Immerse the wafer into HF solution to etch the oxide sacrifice
layer to release Parylene from the substrate.
4. Rinse off and take out the whole Parylene film carefully, and
put on the non-dust cloth to dry.
5. Cut the electrode to suitable shape that contains a cathode and
an anode on a patch with a surgical scissor (see Note 17).
6. Solder wire on the gold pad of microelectrode patch using
silver epoxy adhesive to make tight connection.
7. Store fabricated chip in sealed container.
3.2 Animal
Depilation
The mice are anesthetized by intraperitoneal injection with chloral
hydrate (4% w/v) at a dose of 10 ml/g, and carefully depilated on
the right leg with depilatory cream one day before electroporation
to get a better electroporation results and facilitate observation (see
Note 18).
3.3 Electrical Setup
and Preparation
1. Cover the bench where electroporation will take place with a
piece of foam or other soft materials.
2. Disinfect the 0.5 mm microneedle roller (see Note 19) and
with medicine alcohol, and store in a bio-clean container to dry
for use.
3. Program parameters for BTX ECM830 pulse generator as
follows (see Note 20):
Mode: Low Voltage (LV);
Voltage: 50 V;
Pulse Length: 10 ms;
Number of Pulses: 10;
Interval: 1 s.
4. Connect the flexible microelectrode to pulse generator.
5. Preload tubes with electroporation buffer and medical alcohol.
6. Unfreeze the nucleic acid drugs stored in the centrifuge
microtube.
3.4 Animal Skin
Electroporation
1. Inject the animal intraperitoneally with 4% (w/v) chloral
hydrate (10 ml/g injection volume) to induce anesthesia
15 min before electroporation.
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Dong Huang et al.
Release
1. Remove all of the electroplating mold with PRS 3000 stripper
and rinse off again.
2. Remove the gold seed layer and adhesion promotion layer with
the wet etcher, after that the parallel electrodes are patterned.
The interdigital electrodes are 100 μm wide and 500 μm spaced
(see Note 16).
3. Immerse the wafer into HF solution to etch the oxide sacrifice
layer to release Parylene from the substrate.
4. Rinse off and take out the whole Parylene film carefully, and
put on the non-dust cloth to dry.
5. Cut the electrode to suitable shape that contains a cathode and
an anode on a patch with a surgical scissor (see Note 17).
6. Solder wire on the gold pad of microelectrode patch using
silver epoxy adhesive to make tight connection.
7. Store fabricated chip in sealed container.
3.2 Animal
Depilation
The mice are anesthetized by intraperitoneal injection with chloral
hydrate (4% w/v) at a dose of 10 ml/g, and carefully depilated on
the right leg with depilatory cream one day before electroporation
to get a better electroporation results and facilitate observation (see
Note 18).
3.3 Electrical Setup
and Preparation
1. Cover the bench where electroporation will take place with a
piece of foam or other soft materials.
2. Disinfect the 0.5 mm microneedle roller (see Note 19) and
with medicine alcohol, and store in a bio-clean container to dry
for use.
3. Program parameters for BTX ECM830 pulse generator as
follows (see Note 20):
Mode: Low Voltage (LV);
Voltage: 50 V;
Pulse Length: 10 ms;
Number of Pulses: 10;
Interval: 1 s.
4. Connect the flexible microelectrode to pulse generator.
5. Preload tubes with electroporation buffer and medical alcohol.
6. Unfreeze the nucleic acid drugs stored in the centrifuge
microtube.
3.4 Animal Skin
Electroporation
1. Inject the animal intraperitoneally with 4% (w/v) chloral
hydrate (10 ml/g injection volume) to induce anesthesia
15 min before electroporation.
108
Dong Huang et al.