Biomedical Engineering Reference
In-Depth Information
3
Methods
3.1 ICV Injection
into the Mouse
Brain at Early
Postnatal Days
ICV delivery does not require any extensive surgical devices and can
be easily performed by a single person. However, the small size of
pups at P2-4 and the subsequent small target areas can make the
injection challenging and may require a great deal of practice. Our
procedure requires a sterilized micropipette glass needle that is
attached to a 5 or 10 ml plastic syringe through a long tubing sys-
tem. A fi ber-optic light will be helpful for the visualization of ana-
tomic regions of the brain. A visual demonstration of the ICV
injection technique is published previously by our group [
96
]. Here,
we describe a very detailed protocol of the ICV injection with pho-
tos demonstrating every step of the process (Figs.
6
,
7
,
8
, and
9
).
The needles are pulled from the thin-walled Kimble disposable
borosilicate glass micro capillary pipettes with microfi lament.
Micropipette pulling is performed using P-97 Flaming/Brown
micropipette puller according to the detailed instructions found
in the Sutter Instrument Co. website:
http://www.sutter.com/
3.1.1 Preparation
of the Needles for ICV
Injection
Fig. 6
Proper needles and the proper length of the needle's tip for ICV injection are illustrated. (
a
) Number 19
glass micropipette needles. (
b
) Length of the needle's tip (~5 mm). (
c
) Breaking the needle's tip. (
d
) Length of
the tip, after breaking, for ~2 mm penetration into the mouse cerebral ventricle
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