Assessment of Microthromboembolism After Subarachnoid Hemorrhage - Animal Models of Acute Neurological Injuries: Injury and Mechanistic Assessments - page 370

2. Materials and
Instruments

2.1. Animals

Although the only experience with the assessment of
microthromboembolism at this moment is in a SAH mouse model,
it is certainly possible that the methods can be applied to models of
SAH in rats, dogs, rabbits, and primates ( 8 ).

2.2. Immunohis-
tochemical Staining

To perform the immunohistochemical staining, the following
equipment is required:

0.01 mM sodium citrate (pH 6.0).
0.3% H 2 O 2 .
H 2 O.
10% Normal goat blocking serum.
Phosphate-buffered saline.
Primary antibody to fi brinogen of the species being used for the
animal model available commercially from a variety of sources.
Secondary biotinylated antibody to the species of the primary
antibody.
Avidin biotinylated peroxidase detection kit (e.g., VECTASTAIN

7 ABC Kit, Vector Laboratories, Burlingame, CA).
0.5% Methyl green.

β

2.3. Brain Sectioning
Instruments

To perform the sectioning, the following equipment is required:

Mouse brain slicer matrix, with 1.0 mm coronal slice intervals.
Thin and sharp blades (Leica7 Disposable Microtome Blades, Cat.
63065).

Small beaker or Petri dish.
Water bath.

2.4. Staining
Equipment

Light microscope with digital or charge-coupled device camera
and image capture system.
Ruler.
Blue plastic that provides a good contrast to red and white.
Image analysis system (Imaging Research Inc., St. Catharines,
Ontario, Canada) or Image J (National Institutes of Health imaging
software (NIH, Bethesda, MD).

2.5. Photography
and Processing Tools