Biology Reference
In-Depth Information
2.2.2 Fiber Implants
Implantable fi beroptic connector with metal or ceramic ferrule—
fi ber width: 200
m diameter; fi ber length: according to injec-
tion site DV coordinate (Fig.
1
-bI)
Cannula holder for stereotactic frame
C&B Metabond kit
Acrylic dental cement
μ
Open fi eld chamber for mice: square arena 50 cm × 50 cm made of
gray plastic with 40 cm high walls.
Mouse behavioral tracking system (Viewer3, BIOBSERVE GmbH,
St. Augustin, Germany)
Master-8 pattern generator (A.M.P.I, Jerusalem, Israel)
447 nm, 100 mW diode laser (O.E.M Laser Systems Inc., Salt
Lake City, UT, USA)
Fiberoptic patch-cord (200
2.2.3 Behavioral Testing
μ
m core, Thorlabs Inc., cat # M72L05)
Fiberoptic patch-cord (200
m core, FC/PC to bare ferrule con-
nector, Doric Lenses, Quebec, Canada)
Fiberoptic rotary joint (Doric Lenses; Cat. # FRJ 1X1 FC/FC)
Zirconia sleeve for ferrule-ferrule connection (Thorlabs Inc., cat #
ADAF1)
μ
1. Viral vectors
2.3 Setup
Storage: viruses should be stored in aliquots of 5-10
μ
l at
●
−80 °C.
●
Thawing: on day of surgery, virus aliquots should be
thawed quickly in a gloved hand or a warm water bath and
then kept at 4 °C until use. AAV can be stored after thaw-
ing for several days at 4 °C with minimal loss of activity.
LV is less stable and should be used on the same day.
2. Preparation of surgical tools
●
Set heat bead sterilizer to 250 °C.
●
Sterilize surgical tools by placing them, for at least 45 s, in
sterilizer so that the tips are fully covered by the beads.
●
Arrange the sterilized surgical tools on a sterile surgical
pad next to the stereotaxic instrument and allow them to
cool before beginning the procedure.
3. Preparation of the viral injection syringe
The syringe should be washed thoroughly with DDW and
then sterile saline prior to virus uptake. This is a good
opportunity to make sure that the needle is not clogged by
visually confi rming that liquid is withdrawn and expelled
by slowly moving the syringe plunger up and down.
●
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