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Neuropathology Laboratories
Introduction

The three neuropathology laboratories are divided into:
1. Infarcts and tumors
2. Diffuse anoxic injury, demyelinating diseases, and infection
3. Neurodegenerative and spinal cord diseases.

The laboratories are case based. The clinical and corresponding virtual slides may be accessed using the laboratory links. An explanation of the various stains used on these sections is provided below. Also included in an appendix are seven macroscopic and microscopic normal brain sections to serve as reference material for your examination of the neuropathological slides. These include frontal cortex, two views of the basal ganglia (striatum), hippocampus with temporal lobe, midbrain, pons, and medulla with cerebellum. Due to time constraints, we will not formally cover this material during lab session, but please feel free to ask questions.

NEUROPATHOLOGY STAINS

Luxol - The Luxol fast blue stain, short for Luxol fast blue, cresyl violet and eosin, is really good for staining myelin. Myelin stains blue. Nissl substance and nuclei stain purple, and nearly all else, including the axis cylinders, cytoplasm of the cells, glial fibers, collagen, and red cells, stain pink. (RBCs and collagen may unavoidably stain blue.)

PTAH - This is short for phosphotungstic acid hematoxylin. With this stain, myelin stains blue-purple, nuclei and Nissl substance stain dark purple-blue (basophilic), axis cylinders and cytoplasm stain pink or brown, and glial fibers stain deep blue. Collagen stains scarlet or orange, and RBCs stain blue. This is a really nice stain to show glial fibers.

Hematoxylin and eosin (H&E) - Our workhorse stain. Myelin stains dark pink (eosinophilic), nuclei and Nissl substance blue-purple, and everything else pink. RBCs are reddish orange.

Trichrome - This stain colors myelin orange, nuclei and Nissl substance purple-black, axis cylinders, glial fibers, and other cytoplasm brown-orange. Collagen is stained green and red cells turn orange.

Bielschowsky - This is a gorgeous stain that is used almost exclusively to outline tangles and plaques (which stain black against a gold background). This silver stain colors axis cylinders and neurofibrils black, cytoplasm and nuclei tan to light brown, blood vessels yellow, tan, or brown, and RBCs tan to dark brown. The background or neuropil is a pale tan to yellow (or gold). Glial fibers are usually not stained.

Tau Immunostain - characterizes the accumulation of the microtubule-associated protein tau, which stains a dark brown in the section. Disorders staining positive are known as tauopathies.

Alpha (α) Synuclein Immunostain - characterizes the accumulation of the synaptic-associated protein alpha synuclein, which stains dark brown in the section. Disorders staining positive are known as alpha synucleinopathies.

Αβ-amyloid Immunostain - characterizes the presence of a peptide (Αβ) derived from a fragment of a normal neuronal protein termed APP (amyloid precursor protein), which stains dark brown in the section.

 


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Questions or comments? Dr. Killen: pkillen@umich.edu

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