Tracer Molecule Delivery in the Cisterna Magna: A Method to Deliver Fluorescent Tracer Molecules via Direct Cannulation of the Cisterna Magna in Pig

Abstract

Source: Bèchet, N. B., et al. Direct Cannula Implantation in the Cisterna Magna of Pigs. J. Vis. Exp. (2021).

This video describes the protocol for tracer molecule delivery via direct cannulation of the cisterna magna in a pig brain. This method helps visualize the distribution of cerebrospinal fluid in the glymphatic system.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Cannulation and injection

NOTE: This step requires at least two people and is carried out with the animal's head elevated and neck flexed.

1. Begin with an anesthetized pig, pre-operated with surgically accessible cisterna magna.
2. Ensure that one person elevates and flexes the head and neck of the animal whilst the other palpates for the cisterna magna making a note of its anatomical location.
3. Slowly and carefully introduce....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
18G needle	Mediq
1ml Syringe	FischerSci	15849152
20G cannula	Mediq	NA
22G cannula	Mediq	NA
Bovine serum albumin Alexa-Fluor 647 Conjugate	ThermoFischer	A34785	2 vials (10mg)
CaCl2	Sigmaaldrich	C1016
Dental cement	Agnthos	7508
Glucose	Sigmaaldrich	G8270
Insta-Set CA Accelerator	BSI-Inc	BSI-151
IV line TAP, 3-WAYS with 10cm extension	Bbraun	NA
KCl	Sigmaaldrich	P9333
Marker pen	NA	NA
MgCl2	Sigmaaldrich	M8266
MilliQ water	NA	NA
NaCL	Sigmaaldrich	S7653
NaH2PO4	Sigmaaldrich	S8282
NaHCO3	Sigmaaldrich	S5761
Saline	Mediq	NA
Self-retaining retractors	NA	NA
Superglue	NA	NA
Surgical curved scissors	NA	NA
Surgical towel clamps	NA	NA