In Vivo Imaging of Cerebrospinal Fluid Transport in a Mouse Brain Using Fluorescence Macroscopy

Abstract

Source: Sweeney, A. M., et al. In Vivo Imaging of Cerebrospinal Fluid Transport through the Intact Mouse Skull using Fluorescence Macroscopy. J. Vis. Exp. (2019)

This video demonstrates the process of visualizing cerebrospinal fluid (CSF) flow in a mouse brain using fluorescence imaging. A fluorescent tracer is injected into the cisterna magna (CM), allowing its movement with CSF to be tracked through the subarachnoid and perivascular spaces. Time-lapse imaging captures the tracer's journey, highlighting its distribution into the brain tissue via arterioles.

Protocol

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

1. Preparing the cisterna magna cannula, head plate, and head holder

1. Sterilize all surgical instruments and head plates before surgery.
   NOTE: Fluorescent tracers are delivered directly into CSF via a cisterna magna cannulation.
2. Briefly, using a needle driver, break the tip of a 30G x 12.7 mm (½ inch) needle, ¾ of the way down, and place the blunt end of the needle into one end of polyethylene 10 (PE10) tubing (about 45 cm long). Make sure that only the bevel is protruding o....

Results

Figure 1. Transcranial macroscopic imaging. (A) Schematic of the macrosco.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
0.25% Bupivacaine HCl			University of Rochester Vivarium
100 µL Gastight Syringe Model 1710 TLL, PTFE Luer Lock	Hamilton Company	81020
A-M Systems Dental Cement Powder	Fisher Scientific	NC9991371
Carprofen			University of Rochester Vivarium
Chlorhexidine	Prevantics	B10800
CMOS Camera	Hammamatsu	ORCA Flash 4.0
Head Plate	University of Rochester	No catalog #	Custom made at the machine shop at the University of Rochester
High-Temperature Cautery	Bovie Medical Corporation	AA01
Insta-set Accelerator	Bob Smith Industries	BSI-151
Isoflurane - Fluriso	Vet One	502017	University of Rochester Vivarium
Ketamine			Strong Memorial Hospital Pharmacy
Krazy Glue	Elmer's Products, Inc	No catalog #, see link in comments	https://www.amazon.com/Krazy-Glue-KG48348MR-Advance-Multicolor/dp/B000BKO6DG
Micropore Surgical tape	Fisher Scientific	19-027-761
Paraformaldehyde	Sigma-aldrich	P6148
PE10 - Polyethylene .011" x .024" per ft., 100 ft. continuous	Braintree Scientific	PE10 100 FT
Pump 11 Elite Infusion Only Dual Syringe	Harvard Apparatus	70-4501
PURALUBE VET OINTMENT	Dechra
Puritan PurSwab Cotton Tipped Cleaning Sticks	Fisher Scientific	22-029-553
Research Macro Zoom Microscope	Olympus	MVX10
Simple Head Holder Plate (for mice)	Narishige International USA Inc	MAG-1
Single-use Needles, BD Medical	VWR	BD305106
Sterile Alcohol Prep Pads	Fisher Scientific	22-363-750
Tunable LED	PRIOR Lumen 1600-LED
Xylazine			University of Rochester Vivarium