13
23. Remove
Sample Ring
from the
Clearing Cup
and wash samples with PBSN (PBS with
0.02% sodium azide) in the
Incubation Jar.
Carefully rinse the
Clearing Cup
with distilled
water and store it in its storage solution. It is important to keep the membrane hydrated
at all times.
24. When done clearing samples, or every 10 days (whichever comes first) wash the system
and replace the buffer in the device. We recommend refreshing the Delipidation Buffer
when adding new samples, or every ~3 days, whichever comes first.
25. Wash the device before turning it off to prevent detergent buildup.
Labeling Mode
1. Wash the device and turn off the
Auxiliary Power
.
2. 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.
3. Pour a whole bottle of Sma Primary Device Buffer into the
Chamber.
4. Remove the
Batch Staining Cup
from its storage container, and wash carefully with a
gentle stream of water. (If you are running a single sample experiment use the
Single
Sample Staining Cup
instead).
5. Place the
Batch Staining Cup
into a beaker of distilled water to finish washing it.
6. Remove the
Batch Staining Cup
from the liquid and dump out any water. Carefully use a
kimwipe to soak up any remaining water inside the cup.
7. Rinse out the cup twice with
Primary Sample Buffer.
8. Fill the cup with 40mL of
Primary Sample Buffer
and add the appropriate molecular
probes.
9. Insert the
Sample Ring
into the
Batch Staining Cup
with the samples on their
Mesh
Bags.
Ensure samples are properly pre-incubated.
10. 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.
11. Turn on the
Auxiliary Power.
The buffer should begin to flow and the stirrer should
begin spinning. The cup will initially rotate relatively quickly but will slow down prior to
the experimental run.
12. Close and seal the
Chamber Lid
and the
Case Lid
.
13. Press
“Preset”
until it indicates
Labeling 1 or Labeling 2 Mode
for primary and
secondary antibodies accordingly
.
Change any settings or apply the timer if desired. The
preset settings are the recommended settings.
14. From here, please follow the instructions in the Labeling section of the Full Pipeline
Protocol to finish the experiment.
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