Assessing Blood-Brain Barrier Integrity in a Laser-Induced Brain Injury Rat Model

Abstract

Source: Kuts, R. et al. Laser-Induced Brain Injury in the Motor Cortex of Rats. J. Vis. Exp. (2020)

This video demonstrates assessing blood-brain barrier (BBB) integrity following laser-induced brain injury in rats. A tracking dye is injected into the tail vein, allowing it to circulate to the brain and leak into the tissue through the damaged BBB. The brain is then extracted and processed to release the dye. The fluorescence intensity of the dye is measured to quantify the extent of laser-induced BBB damage.

Protocol

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

1. Laser-induced brain injury experimental procedure

1. Assign 20 rats to a group marked as laser group and 20 rats to another control group (sham-operated).
2. Subject the laser group rats to laser irradiation at 50J X 10 points in the following ma....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
50% trichloroacetic acid	SIGMA - ALDRICH	76-03-9
Brain & Tissue Matrices	SIGMA - ALDRICH	15013
Cannula Venflon 22 G	KD-FIX	1.84E+11
Centrifuge Sigma 2-16P	SIGMA - ALDRICH	Sigma 2-16P
Compact Analytical Balances	SIGMA - ALDRICH	HR-AZ/HR-A
Digital Weighing Scale	SIGMA - ALDRICH	Rs 4,000
Dissecting scissors	SIGMA - ALDRICH	Z265969
Eppendorf pipette	SIGMA - ALDRICH	Z683884
Eppendorf Tube	SIGMA - ALDRICH	EP0030119460
Ethanol 96 %	ROMICAL		Flammable Liquid
Evans Blue 2%	SIGMA - ALDRICH	314-13-6
Fluorescence detector	Tecan, Männedorf Switzerland	model Infinite 200 PRO multimode reader
Heater with thermometer	Heatingpad-1	Model: HEATINGPAD-1/2
Infusion Cuff	ABN	IC-500
Isofluran, USP 100%	Piramamal Critical Care, Inc	NDC 66794-017
Multiset	TEVA MEDICAL	998702
Olympus BX 40 microscope	Olympus
Petri dishes	SIGMA - ALDRICH	P5606
Scalpel blades 11	SIGMA - ALDRICH	S2771
Sharplan 3000 Nd:YAG (neodymium-doped yttrium aluminum garnet) laser machine	Laser Industries Ltd
Stereotaxic head holder	KOPF	900LS
Sterile Syringe 2 ml	Braun	4606027V
Syringe-needle 27 G	Braun	305620