F
LEXCELL
®
I
NTERNATIONAL
C
ORPORATION
21
end. Be careful not to stimulate or crush any
cells on the slide.
13.
Place the Culture Slip
®
into one of the slots
in the Streamer
®
device.
Be sure that the
side with cells attached is facing the flow
area (the shorter slot parallel and adjacent
to the slide slot).
Be careful not to chip the
glass against the stainless steel surface.
14.
Repeat this for the other Culture Slips
®
,
making sure the surface with the cells all
face the proper direction in the flow device.
NOTE:
All six slots must be filled to ensure proper
flow rate readings. If you do not wish to use all six
Culture Slips
®
with cells, use blank Culture Slips
®
for the remaining slots.
15.
Once all Culture Slips
®
are in the Streamer
®
unit, close the lid and tighten the screws
using the hex head tool provided with the
system.
NOTE:
As you are moving the Streamer
®
from this
point on, always position the device vertically such
that the inlet connector is at the bottom and the
outlet connector is at the top.
If you wish to run the Streamer
®
on its side, first fill
the remainder of the system with fluid so that all air
is completely out of the Streamer
®
chamber.
Be aware that any air that accidentally enters the
system may eventually form a dry area at the
topmost slide so that these cells will no longer see
fluid media and shear stress.
Be sure that your system does not regularly see
additional air bubbles (after initial filling and air
bubble elimination) before using the Streamer
®
on
its side.
16.
Move the tray with the system components
back to the incubator. Put the Phar-Med
®
tubing back into the MasterFlex
®
pump
head and clamp the head down.
17.
Unscrew the clamp on the Phar-Med
®
tubing to open the flow path to full capacity.
18.
If you are running the experiment manually,
set the pump to the desired flow rate and
press the start button. If running the
experiment under software control, go to
the
System
tab, click on the
Configure
button, assign the
User
,
Regime
and
Apparatus
. Click
Update
, then
Start
.
19.
Once the experiment is over, move the
Streamer
®
back to the tissue culture hood
and remove the slides.
20.
Once the slides are removed, place the
Streamer
®
back into the incubator and run
deionized water through the system to
remove all remaining media. Refresh the
deionized water and run a second or third
time if necessary.
Be sure to never leave the
Streamer
®
with culture media inside as
this will corrode the stainless steel finish
over time.
F
ILLING THE
S
YSTEM TO
E
LIMINATE
A
IR
B
UBBLES
Before using the Streamer
®
system with cells,
all of the tubing must be filled with media and
all air bubbles removed. To fill the system,
create a regime with two steps, the first in FWD
mode and the second in REV mode. Each step
should be 2 minutes at a shear stress level ½ that
of which your device is capable. This will give
sufficient time for fluid to fill all of the tubing.
As you notice air bubbles in the silicone tubing
at different locations, shake the tubing to release
the air bubbles.
P
OST
-E
XPERIMENT
A
NALYSIS
Upon removal of the slides from the flow
device, many post-flow evaluations can be
done.
Cells on the Culture Slips
®
can be fixed
with formalin then permeabilized and
stained with rhodamine Phalloidin and
DAPI to visualize cell alignment.
