Visualization of a Cerebral Thrombus in a Mouse Using Micro-Computed Tomography Imaging

Abstract

Source: Kim, D., et. al. Combined Near-infrared Fluorescent Imaging and Micro-computed Tomography for Directly Visualizing Cerebral Thromboemboli. J. Vis. Exp. (2016)

This video demonstrates visualization of cerebral thrombus in mice using micro-computed tomography (micro-CT) imaging. A cerebral thrombus mouse model is injected with fibrin-targeted gold nanoparticles acting as a contrasting agent. The mouse is placed on the bed of a micro-CT machine, and images are acquired from multiple angles. Using appropriate software, images are reconstructed and processed.

Protocol

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

1. Preparation of Exogenously Formed Clot Labeled with Fluorescence Marker (Figure 1)

1. Anesthetize a mouse in an induction chamber using 3% isoflurane mixed with 30% oxygen (1.5 L/min 100% oxygen). Ensure adequate depth of anesthesia by observing muscle tone and confirming the absence of the toe pinch reflex.
2. Place the animal on a sterile drape in a prone position and keep it under anesthesia using an inhalation mask and 2% isoflurane mixed with 30% oxygen. Perform the following procedures using aseptic te....

Results

Figure 1: Overview of the Preparation of Fluorescently Labeled Blood Clot. (A) Mixing of fresh.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Machines
microCT	NanoFocusRay, JeonJu, Korea	NFR Polaris-G90
Laser Doppler flowmeter	Perimed, Stockholm, Sweden	PeriFlux System 5000
Surgical microscope	Leica Microsystems, Seoul, Korea	EZ4HD
Inhalation anesthesia machine	PerkinElmer, Massachusetts, USA	XGI-8
Software
NFR control	NanoFocusRay, JeonJu, Korea	NFR Polaris-G90	microCT control software
Lucion	Infinitt, Seoul, Korea	Lucion	3D render imaging software
Lab chart 7	ADInstruments, Colorado, USA	Lab chart 7	rCBF
Image J software	Wanye Rasband, NIH, USA	1.49d	imaging analysis
Devices/Instruments
Infusion pump	Harvard, Massachusetts, USA	pump 22(55-2226)
Homeothermic blanket	Panlab, Barcelona, Spain	HB101
Pocket cautery	Daejong, Seoul, Korea	DJE-39
Brain matrice	Ted pella, CA, USA	15003	coronal section
PE-50 tubing	Natsume, Tokyo, Japan	SP-45(PE-50)	I.D. 0.58 mm O.D. 0.96 mm
PE-10 tubing	Natsume, Tokyo, Japan	SP-10(PE-10)	I.D. 0.28mm O.D. 0.61 mm
30 gauge needle	sungshim-medical, Seoul, Korea
Syringe	CPL-medical, Ansan, Korea		1 & 3 cc
Gauze	Panamedic, Cheonan, Korea
Tape	Scotch, Seoul, Korea	3M-810
Micro forceps	Fine Science Tools, Vancouver, Canada	11253-27	Dumont #L5
Micro scissor	Fine Science Tools, Vancouver, Canada	15000-03	Vannas spring
Scissor	Fine Science Tools, Vancouver, Canada	14084-08	8.5 cm
Black silk suture	Ailee, Busan, Korea	SK6071, SK728	6-0 and 7-0
Reagents
Meloxicam	Yuhan, Seoul, Korea
Vet ointment	Novartis, Basel, Swiss
10% Povidone-iodine (betadine)	Firson, Cheon-an, Korea
Ferric chloride	Sigma, Missouri, United States	157740-5G
TTC	Amresco, Ohio, USA	0765-100g
Isoflurane	Hana-Pham, Gyeonggi, Korea	Ifran	100 mL
PBS	Welgene, Daegu, Korea	LB001-02	500 mL
Gold nanoparticles			Synthesis
Normal saline	Daihan Pham, Seoul, Korea	48N3AF3	20 mL