12
Operation
Clearing Mode
For more detailed information regarding sample preparation, please refer to the Full Active
Pipeline Protocol. This will show the basic information on running the system to delipidate
samples.
1. Insert SHIELD fixed samples in the
Mesh Bags
, taking note of which sample is in which
bag. When inserting mouse brains into the bags put the cerebellum facing down to avoid
damaging olfactory bulbs.
2. Place the
Sample Ring
on the
Ring Stand
.
3. Hang the
Mesh Bags
on the
Sample Ring
and secure them with the
Lock Ring.
4. Place
Ring Stand
in
Incubation Jar
and incubate samples in Delipidation Buffer overnight
at RT.
5.
and turn off the
Auxiliary Power.
6. Drain out any liquid from the device and ensure the
Drainage Valve
is closed. Soak up
any remaining liquid from the
Chamber
using a paper towel.
7. Pour a whole bottle of Sma Conduction Buffer into the
Chamber
.
8. Remove the
Clearing Cup
from its storage container, and wash carefully with a gentle
stream of water.
9. Place the
Clearing Cup
into a beaker of distilled water for several minutes to finish
washing the cup.
10. Remove the
Clearing Cup
from the liquid and dump out any water. Carefully use a
kimwipe to soak up any remaining water inside the cup.
11. Fill the cup with 32mL of Delipidation Buffer.
12.
13. Place the
Sample Ring
on top of the
Clearing Cup
with the samples inside. Add more
Delipidation Buffer if the samples are not fully submerged.
14. Insert the cup in the
Chamber
, lining up the pegs on the bottom with those in the
Chamber Hex Piece.
The cup will fit in snugly.
15. Turn on the
Auxiliary Power.
16. Close the
Chamber Lid
and secure it with the thumbscrew.
17. Close the
Case Lid
.
18. Press
“Preset”
until it indicates
Clearing Mode.
Change any settings or apply the timer if
desired. The preset settings are the recommended settings for mouse brains.
19. Turn on the
Electrophoresis Power
.
20. Wait for your samples to clear.
21. Once fully delipidated, turn off
Electrophoresis Power
and
Auxiliary Power
.
22. Open the
Case Lid
and
Chamber Lid
and remove the
Clearing Cup
from the
Chamber.
www.lifecanvastech.com
