Inducing Blood Clot Formation in the Mouse Brain Using a Photosensitive Dye

Samenvatting

Source: Talley Watts, L., et al. Rose Bengal Photothrombosis by Confocal Optical Imaging In Vivo: A Model of Single Vessel Stroke. J. Vis. Exp. (2015)

This video demonstrates a technique to induce blood clot formation in a mouse brain. An anesthetized mouse with a cranial window and a thinned skull undergoes photothrombosis through the activation of a photosensitive dye using high-intensity light. This process generates reactive oxygen species, which damage the vessel walls and trigger clot formation, ultimately obstructing blood flow to the brain.

Protocol

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

1. Anesthetizing for Cortical Preparation

1. Place the mouse in an induction chamber with 2-3% isofluorane mixed with oxygen to induce anesthesia. Observe the respiration rate decrease as the mouse is induced. Pinch the paw of the mouse to determine whether the mouse is ready to move to the nose cone. Note: Anesthe....

Resultaten

Figure 1: Stainless Steel Ring. Three views (top, side and bottom views) are shown of the stainless-steel ring holder that is applied to the skull of the mouse to affix it to the stereotaxic holder.

Materialen

Lijst van materialen gebruikt in dit artikel

Naam	Bedrijf	Catalogusnummer	Opmerkingen
Reagents
Rose Bengal	Sigma	330000
Isoflurane Anesthetic	MWI Veterinary Supply	088-076
Vetbond	1469SB	1469SB
aCSF	126 mM NaCl, 2.5 mM KCl, 1.25 mM NaH2PO4, 2 mM MgCl2, 2 mM CaCl2, 10 mM glucose and 26 mM NaHCO3 (pH 7.4).
Equipment
Dissecting Scissors	Bioindustrial Products	500-410
Operating scissors 14 cm	Bioindustrial Products	12-055
Forceps Dumont High Tech #5 style, straight	Bioindustrial Products	TWZ-301.22
LabJack 132X80	Optosigma Co	123-6670
Platform for Labjack 8X 8	Optosigma Co	145-1110
Ear bar holder from stereotaxic setup	Stoelting/Cyborg	51654
Dispomed Labvent Rodent anesthesia machine	DRE, Inc.	15001
Tech IV Isoflurane vaporizer	DRE, Inc.	34001
F Air Canister	DRE, Inc	80120
Bain circuit breathing tube	DRE, Inc	86111B
Rodent adapter for bain tube	DRE, Inc	891000
O2 regulator for oxygen tanks	DRE, Inc	CE001E
Rodent induction chamber	DRE, Inc	15004C
Ethicon Silk 6-0; 18 in with P-3 needle	Suture Express	1639G
Objective inverter Optical Adapter	LSM technologies
Foredom drill Dual voltage 110/120	Foredom	134.53