Difference between revisions of "ICV Cannulation"

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(added dental acrylic)
(Update Bosworth link to Keystone.)
 
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[[File:ICV cannulation.png|thumb|Cross-section schematic of cannula attached to skull with dental acrylic and screws.]]
 +
For other stereotaxic coordinates, see also [[Amygdala]], [[JA Lateral Hypothalamus Lesions]].
  
−
See also [[Amygdala]] for other cannulation coordinates.
+
==Lateral Ventricular Cannula==
−
 
 
−
==Lateral Ventricular Cannula:==
 
  
 
1.2 mm caudal to bregma
 
1.2 mm caudal to bregma
Line 11: Line 11:
 
4 mm below the skull surface
 
4 mm below the skull surface
  
−
===Method===
 
  
−
Each rat was stereotaxically implanted with a 22 gauge, stainless steel, guide
+
Under isoflurane anesthesia, each rat was stereotaxically implanted with a 22-gauge, stainless steel, guide
−
cannula (Plastics One, Roanoke, VA) aimed towards the
+
cannula (model C313G, Plastics One, Roanoke, VA) aimed towards the lateral cerebral ventricle (1.2 mm caudal to bregma, 1.5 mm
−
lateral cerebral ventricle (1.2 mm caudal to bregma, 1.5 mm
+
lateral to the midline, and 4 mm below the skull surface; Houpt 1998 PMID 9437750). Guide cannulas were held in place with dental acrylic bonded to
−
lateral to the midline, and 4 mm below the skull surface).
+
stainless steel screws anchored to the skull. A 28-gauge obdurator was inserted into each guide cannula and remained in place
−
Guide cannulas were held in place with dental acrylic
+
except during injections when it was removed and replaced with an 28-gauge injector that extended 1.0 mm beyond the tip of the guide cannula.
−
bonded to stainless steel screws anchored to the skull. An
+
 
−
obdurator was inserted into each guide cannula and remained
+
To test the patency and placement of the cannula in the lateral ventricle,  water-replete rats were injected with
−
in place except during injections when it was removed
+
100 ng human angiotensin II (Ang II; Sigma Chemical Co, St Louis, MO) dissolved in 5 µl volume of saline (0.15 M).
−
and replaced with an injector that extended 1.0 mm
+
The volume of all icv injections was 5 µl, delivered over 30s with a handheld 50 µl syringe (Hamilton Co, Reno, NV).
−
beyond the tip of the guide cannula.
+
Immediately after the ICV injection, rats were returned to their home cage with access to a water bottle.
 +
The latency to drink was recorded; rats that failed to drink within 2 minutes of the angiotensin II injection were
 +
dropped from the study. Cannula placements were also verified postmortem by sectioning through the brain.
 +
ICV injections of 5 µl  volume were given by hand to lightly restrained rats using a 50 µl Hamilton microsyringe.
 +
 
 +
==Fourth Ventricular Cannula==
 +
 
 +
3.2 mm caudal to lambda
 +
 
 +
0 mm lateral to the midline
 +
 
 +
7.2 mm below the skull surface
 +
 
 +
Under isoflurane anesthesia, each rat was stereotaxically implanted with a single 22-gauge stainless steel guide cannula
 +
(model C315G; Plastics One, Inc., Roanoke, VA) aimed towards the fourth ventricle (4V).  The tip of the guide cannula
 +
was positioned on the midline 3.2 mm caudal to lambda and 7.2 mm below the skull surface (stereotaxic coordinates
 +
taken from (Baird et al. 2009 PMID 19008313)). Guide cannulas were held in place with dental acrylic bonded to stainless
 +
steel screws anchored to the skull. A 28-gauge obdurator extending 1.0 mm beyond the guide
 +
cannula was inserted into each guide cannula and remained in place except during injections when it was removed and
 +
replaced with a 28-gauge injector that extended 1.0 mm beyond the tip of the guide cannula.
 +
 
 +
To test the patency and placement of the cannula in the lateral ventricle,  water-replete rats were injected with
 +
XXX ng bombesin (California Peptides) dissolved in 5 µl volume of saline (0.15 M). The volume of all icv injections
 +
was 5 µl, delivered by hand over 30s to lightly restrained rats using a  50 µl Hamilton microsyringe (Hamilton Co, Reno, NV). 
 +
Immediately after the ICV injection, rats were returned to their home cage with access to a water bottle.
 +
The latency to initiate and duration of grooming was recorded; rats that failed to groom within 2 minutes of the
 +
bombesin injection or groomed for less than XXX minutes were dropped from the study. Cannula placements were
 +
also verified postmortem by sectioning through the brain.
  
 
==Supplies==
 
==Supplies==
  
−
New Truliner Denture Lining  Standard Kit, Pink [http://www.bosworth.com/product_detail.asp?PRD_ID=18 Bosworth #0921970] $45.00
+
===Plastics One Cannulas and Screws===
 +
 
 +
:[https://www.p1tec.com P1 Technologies (née Plastics One)]
 +
:6591 Merriman Rd
 +
:Roanoke VA 24018
 +
:Tel: 540-772-7950
 +
:Fax: 540-777-5900
 +
 
 +
''Check prices on annual price list.''
 +
 
 +
====22 gauge guide cannulas for ventricular injections====
 +
 
 +
Use C313 components, which provide 28 gauge dummy and injectors.
 +
e.g., for guide cannula(G) projecting 5 mm below skull surface,
 +
with large-cap dummy cannulas (DC) and injectors (I) projecting 1 mm beyond tip of the guide cannula:
 +
 
 +
:C313G/SPC 5 mm
 +
:C313DC/SPC(LG) for 5 mm C313G with 1 mm projection
 +
:C313I/SPC for 5 mm C313G with 1 mm projection
 +
 
 +
====26 gauge guide cannulas for site-specific injections====
 +
 
 +
Use C315 components, which provide 33 gauge dummy and injectors.
 +
e.g., for guide cannula(G) projecting 8 mm below skull surface,
 +
with small-cap dummy cannulas (DC) and injectors (I) projecting 0.5 mm beyond tip of the guide cannula:
 +
 
 +
:C315G/SPC 8 mm
 +
:C315DC/SPC(SM) for 8 mm C315G with 0.5 mm projection
 +
:C315I/SPC for 8 mm C315G with 0.5 mm projection
 +
 
 +
====Screws for skull====
 +
 
 +
We use screws of 0-80 size with shaft length 3/32 inches (2.4 mm).
 +
These are a little long, but they are stronger and easier to work with than  1/16 inch (1.6 mm) long screws.
 +
Also need the small drill bit.
 +
 
 +
:0-80 X 3/32 (packages of 100)
 +
:D#56 1.18 mm Drill Bit for 0-80 screws
 +
 
 +
 
 +
----
 +
 
 +
===Other supplies needed - more detail to come===
 +
 
 +
====Pin Vise for hand drilling the screws====
 +
 
 +
Most pin vises are nickel plated, and so cannot be autoclaved but must be sterilized e.g. with 70% EtOH.
 +
:VWR 63041-530 Pin Vise - Double-Ended, stainless steel
 +
 
 +
Little tubing for washer around top of guide cannula?
 +
 
 +
 
 +
Superglue
 +
 
 +
Size of screwdriver? source of SS screw drivers?
 +
 
 +
Trephin drill bits
 +
 
 +
Hamilton microsyringe and needle
 +
 
 +
PE-50 tubing to connect with injector
 +
 
 +
Syringe Pump
 +
 
 +
 
 +
====New Truliner Denture Lining  Standard Kit, Pink====
 +
 
 +
From either Henry Schein Dental, #2502527EZ, $105.29, or,
 +
[https://dental.keystoneindustries.com/product/new-truliner/ Bosworth #0921970] $45.00
  
 
:Harry J. Bosworth Company
 
:Harry J. Bosworth Company
Line 37: Line 131:
  
  
−
[[Category:Taste_Aversion]]
+
[[Category:Taste_Aversion]][[Category:Protocols]]

Latest revision as of 08:05, 20 September 2022

Cross-section schematic of cannula attached to skull with dental acrylic and screws.

For other stereotaxic coordinates, see also Amygdala, JA Lateral Hypothalamus Lesions.

Lateral Ventricular Cannula

1.2 mm caudal to bregma

1.5 mm lateral to the midline

4 mm below the skull surface


Under isoflurane anesthesia, each rat was stereotaxically implanted with a 22-gauge, stainless steel, guide cannula (model C313G, Plastics One, Roanoke, VA) aimed towards the lateral cerebral ventricle (1.2 mm caudal to bregma, 1.5 mm lateral to the midline, and 4 mm below the skull surface; Houpt 1998 PMID 9437750). Guide cannulas were held in place with dental acrylic bonded to stainless steel screws anchored to the skull. A 28-gauge obdurator was inserted into each guide cannula and remained in place except during injections when it was removed and replaced with an 28-gauge injector that extended 1.0 mm beyond the tip of the guide cannula.

To test the patency and placement of the cannula in the lateral ventricle, water-replete rats were injected with 100 ng human angiotensin II (Ang II; Sigma Chemical Co, St Louis, MO) dissolved in 5 µl volume of saline (0.15 M). The volume of all icv injections was 5 µl, delivered over 30s with a handheld 50 µl syringe (Hamilton Co, Reno, NV). Immediately after the ICV injection, rats were returned to their home cage with access to a water bottle. The latency to drink was recorded; rats that failed to drink within 2 minutes of the angiotensin II injection were dropped from the study. Cannula placements were also verified postmortem by sectioning through the brain. ICV injections of 5 µl volume were given by hand to lightly restrained rats using a 50 µl Hamilton microsyringe.

Fourth Ventricular Cannula

3.2 mm caudal to lambda

0 mm lateral to the midline

7.2 mm below the skull surface

Under isoflurane anesthesia, each rat was stereotaxically implanted with a single 22-gauge stainless steel guide cannula (model C315G; Plastics One, Inc., Roanoke, VA) aimed towards the fourth ventricle (4V). The tip of the guide cannula was positioned on the midline 3.2 mm caudal to lambda and 7.2 mm below the skull surface (stereotaxic coordinates taken from (Baird et al. 2009 PMID 19008313)). Guide cannulas were held in place with dental acrylic bonded to stainless steel screws anchored to the skull. A 28-gauge obdurator extending 1.0 mm beyond the guide cannula was inserted into each guide cannula and remained in place except during injections when it was removed and replaced with a 28-gauge injector that extended 1.0 mm beyond the tip of the guide cannula.

To test the patency and placement of the cannula in the lateral ventricle, water-replete rats were injected with XXX ng bombesin (California Peptides) dissolved in 5 µl volume of saline (0.15 M). The volume of all icv injections was 5 µl, delivered by hand over 30s to lightly restrained rats using a 50 µl Hamilton microsyringe (Hamilton Co, Reno, NV). Immediately after the ICV injection, rats were returned to their home cage with access to a water bottle. The latency to initiate and duration of grooming was recorded; rats that failed to groom within 2 minutes of the bombesin injection or groomed for less than XXX minutes were dropped from the study. Cannula placements were also verified postmortem by sectioning through the brain.

Supplies

Plastics One Cannulas and Screws

P1 Technologies (née Plastics One)
6591 Merriman Rd
Roanoke VA 24018
Tel: 540-772-7950
Fax: 540-777-5900

Check prices on annual price list.

22 gauge guide cannulas for ventricular injections

Use C313 components, which provide 28 gauge dummy and injectors. e.g., for guide cannula(G) projecting 5 mm below skull surface, with large-cap dummy cannulas (DC) and injectors (I) projecting 1 mm beyond tip of the guide cannula:

C313G/SPC 5 mm
C313DC/SPC(LG) for 5 mm C313G with 1 mm projection
C313I/SPC for 5 mm C313G with 1 mm projection

26 gauge guide cannulas for site-specific injections

Use C315 components, which provide 33 gauge dummy and injectors. e.g., for guide cannula(G) projecting 8 mm below skull surface, with small-cap dummy cannulas (DC) and injectors (I) projecting 0.5 mm beyond tip of the guide cannula:

C315G/SPC 8 mm
C315DC/SPC(SM) for 8 mm C315G with 0.5 mm projection
C315I/SPC for 8 mm C315G with 0.5 mm projection

Screws for skull

We use screws of 0-80 size with shaft length 3/32 inches (2.4 mm). These are a little long, but they are stronger and easier to work with than 1/16 inch (1.6 mm) long screws. Also need the small drill bit.

0-80 X 3/32 (packages of 100)
D#56 1.18 mm Drill Bit for 0-80 screws


Other supplies needed - more detail to come

Pin Vise for hand drilling the screws

Most pin vises are nickel plated, and so cannot be autoclaved but must be sterilized e.g. with 70% EtOH.

VWR 63041-530 Pin Vise - Double-Ended, stainless steel

Little tubing for washer around top of guide cannula?


Superglue

Size of screwdriver? source of SS screw drivers?

Trephin drill bits

Hamilton microsyringe and needle

PE-50 tubing to connect with injector

Syringe Pump


New Truliner Denture Lining Standard Kit, Pink

From either Henry Schein Dental, #2502527EZ, $105.29, or, Bosworth #0921970 $45.00

Harry J. Bosworth Company
7227 North Hamlin Avenue
Skokie, Illinois 60076 USA
Toll-Free: 800-323-4352
Phone: 847-679-3400
Fax: 847-679-2080
E-Mail: HJBinfo@bosworth.com
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