Light Microscopic Assessment - Animal Models of Acute Neurological Injuries: Injury and Mechanistic Assessments - page 458

Xylene: harmful for inhalation and skin contact; has neurotoxicity.
PBS (Phosphate-Buffered Saline).

2.2. For HE Staining

Graded Ethanol (70%, 80%, 90%, 99.5%, 100%).
Xylene: harmful for inhalation and skin contact; has neurotoxicity.
PBS (Phosphate-Buffered Saline).
Hematoxylin.
Eosin.
0.25% Acid Ethanol: 95% ethanol 2578 ml, dH 2 O 950 ml,
HCl 9 ml.

Weigert's Resorcin-fuchsin solution.
Weigert's Ferric Hematoxylin.
Van Gieson solution.
Graded ethanol.

2.3. For Elastica van
Gieson Staining

As an immunohistological assessment, an enzyme antibody tech-
nique is mainly used in the research of cerebral vasospasm. There
are many methods, such as LAB (Linked Avidin-Biotin) method,
ABC (Avidin-Biotin Complex) method, TSA (Tyramide Signal
Amplifi cation) method, PAP (Peroxidase-anti Peroxidase Complex)
method, LSAB (labeled streptavidin) method, and labeled polymer
method. ABC method has been conventional; however, LSAB and
labeled polymer methods are recently prevailing because of its
simplicity and high sensitivity. Most of the materials are included in
the commercially available kit.

2.4. For
Immunostaining

3. Methods

For HE and elastica van Gieson staining, the specimen taken after
the perfusion fi xation is postfi xed in the same solution as the perfu-
sion fi xation. The specimen usually consists of the artery to be
aimed for and the neighboring brain. For a representative, the basi-
lar artery with the pons is the study frequently. After that, the spec-
imen is washed by a current overnight, dehydrated by a graded
ethanol, and substituted in xylene. The specimen is embedded in
paraffi n (Fig. 1 and see Note 1).
The embedded specimen is sliced using a microtome on thick-
ness of 4-6

3.1. Conventional
Staining

m, the slice is mounted on a glass, and it is deparaf-
fi nized with xylene, rehydrated with a graded ethanol, and rinsed
in distilled water.

μ

1. HE staining (Fig. 2a )

HE staining has been most frequently used to evaluate the
degree of cerebral vasospasm. Hematoxylin stains cell nuclei