IP Icilin Behavior Protocolx - storylab

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14 Δεκ 2013 (πριν από 3 χρόνια και 10 μήνες)

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Icilin Behavior Protocol

(Revised 8/2/2010)


Get

the

mice and bring them to behavior room.


Weigh the mice and record their weights. To hel
p
you

identify the mice

later on, use a sharpie
and mark their tails. For example, mouse #2 has two marks on its tail.


Leave the mice in the behavior room and let them acclimate for 2 hours with white noise in the
background.


While
the
mice are acclimating, suspend icilin in
1% Tween80 in PBS.



(Skip this step if you already have 1% Tween)
To make 1% Tween80

in PBS
, make a
large volume in a 50mL conical

and vortex until Tween is completely dissolved
.

For
example, to make 40mL of the solution, add 39.6mL of PBS and 400
μ
L Tween8
0 into
the conical

and vortex well
.



Based on the total weight

of the mice, determine how much icilin and 1% Tween80 you
will need.
The following formulas will make a







icilin solution
(multiplying
by 1.2 adjusts for

error)
:



Icilin

















(

)








1% Tween
















(

)








Add the

amount of icilin and 1% Tween
that you just calculated
into an eppendorf tube
and vortex. Icilin doe
s NOT

need to be
comple
tely
dissolved.



Use parafilm
and

wrap the tube.



Sonicate the
icilin
mixture for 2 hours.

The sonicator is set only to 60 minutes
, so you
will need to start it again after an hour
. Before you
start the second hour of sonication
,
take out the icilin
mixture and vortex it lightly
.


After 2 hours of acclimating the mice, take the mice out of their cage
s

and acclimate them in the
clear
behavior
box
es

for 20 minutes.


When ready to continue, inject the appropriate amount of icilin solution into each mouse
. To
calculate how much icilin to use, follow this formula:











(

)








Each unit on the syringe is 10
μ
L. For example, if you need to inject 90
μ
L, you will need
9 units.

When doing an intraperitoneal injection, make

sure the hole in the syringe is facing up. Go in at
about a 30
o

angle and aspirate (suck up) to make sure there are no colored fluids. If it is clean,
inject the icilin and place the mouse back in
to

the behavior box.

Make sure to put a piece of
paper i
n between the
behavior
boxes so the mice cannot see each other.



In between each experiment, clean up any mouse droppings
.



Stack two boxes on top of each other to prevent the mice from jumping out.


Observe the mice for 20 minutes

with sunglasses on.

Record

the number of jumps and wet
-
dog
shakes every 5 minutes.


After the experiments,
clean the boxes
and lids
thoroughly with the citrus spray and rinse them
off with the water spray. Clean the experiment surface in the same manner. Clean the desk with

ethanol.



At the end of every week, clean the boxes and lids with soap and water. Let them air dry
over the weekend.