Light Microscopic Assessment - Animal Models of Acute Neurological Injuries: Injury and Mechanistic Assessments - page 459

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Pons

Basilar artery

90 C

Fig. 1. In order to slice the artery in a right angle in its axis, the specimen should be
embedded in a right angle on the plate.

blue, while eosin stains cytoplasm, connective tissue and other
extracellular substances pink or red.
(a) Hematoxylin staining: 3-6 min × 1
(b) Rinse in running tap water: 10-20 min × 1
(c) Eosin staining: 2-30 min × 1
(d) Dehydrated in graded ethanol
(e) Xylene
2. Elastica van Gieson staining (Fig. 2b )

Elastica van Gieson staining has been used to evaluate the
change of internal elastic lamina of arterial wall. This method
stains elastic fi bers blue-black to black, cell nuclei blue to black,
collagen red, and other tissue elements yellow.
(a) Weigert's Resorcin-fuchsin solution: 30 min to 12 h
(b) 100% alcohol: 3 min × 3
(c) Wash in water
(d) Weigert's Ferric hematoxylin solution: 5-10 min
(e) Wash in water: 10 min
(f) Stain in van Gieson: 2-5 min
(g) Differentiated in acid alcohol (1% HCl 70% alcohol)
(h) Dehydrated in graded ethanol
(i) Xylene

For immunostaining, production of the specimen is usually divided
as shown in Fig. 3 (see Notes 2 and 3). When the specimen is taken
without perfusion fi xation, it is embedded in optimum cutting
temperature compound, and frozen. The block is sectioned using

3.2. Immunostaining
(Fig. 2c )