Detection of Cerebral Microhemorrhages in a Rat Model

Abstract

Source: Li, D., Zhào, H., et al. Sub-acute Cerebral Microhemorrhages Induced by Lipopolysaccharide Injection in Rats. J. Vis. Exp. (2018).

This video demonstrates an assay to detect Cerebral microhemorrhages (CMHs) in a rat model. CMHs are brain bleeds due to blood-brain barrier disruption, identified by leaked red blood cells (RBCs). In a rat model, fixed brain tissue is cryoprotected, sectioned, and stained with hematoxylin-eosin for RBCs, allowing microscopic visualization of CMHs. Leaked RBCs appear as red-orange under the microscope.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

1. Materials

1. Preparation of lipopolysaccharide (LPS) injection
   1. Add 25 mL of distilled water to 25 mg of LPS powder derived from Salmonella typhimurium to a final concentration of 1 mg/mL. Store the injection in a sterile tube at 4 °C.
      CAUTION: LPS is toxic.
   2. Prepare a 2% Evans blue (EB) solution in normal 0.9% saline solution to keep EB injection at working concentration.

2. A....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
LPS	Sigma-Aldrich	L-2630	for inflammation induction
EB	Sigma-aldrich	E2129	for EB leakage detection
HE staining	Solarbio	G1120	Kit for HE staining
chloral hydrate	Sigma-Aldrich	47335U	For anesthesia
phosphate buffer saline (PBS)	Solarbio	P1022	a kind of buffer solution commonly used in experiment
0.9% saline solution	Hainan DonglianChangfu Pharmaceutical Co., Ltd., China		solution for perfusion
paraformaldehyde	Sigma-Aldrich	158127	a kind of solution commonly used for fixation
20% sucrose solution	Solarbio	G2461	a kind of solution commonly used for fixation
30% sucrose solution	Solarbio	G2460	a kind of solution commonly used for fixation
vet ointment	Solcoseryl eye gel, Bacel, Switzerland		for rat's eyes protection