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Neuropathological examination
Anna Oevermann Division of Neurological Sciences (Neurocenter) Vetsuisse Faculty, University of Bern
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Nuts and bolts of neuropathology
Removal of brain and spinal cord
Neuroanatomy
Macroscopic examination
Histology sampling
Basic reaction patterns at the histological level
Recognizing the major patterns
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ESAVS 2014
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http://www.vetsuisse-bern.ch/~vet-iml/lernmodule/htmls/npintro.html?neuropatho%7Cnpintro
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Removal of the brain
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Opening of the skull
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Removal of the spinal cord
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Neuroanatomy Use the ventricular system as your road map!
lateral ventricle
V. bulbi olfactorii
III.
IV.
aquaeduct
central canal olfactory lobe
cerebral cortex
thalamus
midbrain
cerebellum
spinal cord
pons, medulla
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olfactory lobe
cerebral cortex
thalamus midbrain
pons, medulla, cerebellum
spinal cord
III. IV.
lateral ventricle
aquaeduct
central canal V. bulbi olfactorii
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cerebral cortex caudate nucleus
rostral commissure
optic chiasm
capsula interna
claustrum putamen globus pallidus
corpus callosum
cerebral white matter
olfactory bulb olfactory tract
lateral ventricle
rhinal fissure
septal nuclei
transversal sections
Frontal lobe Basal nuclei
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thalamus
hypothalamus
caudate nucleus
amygdala optic tract
corpus callosum
fimbria
capsula interna
hippocampus
hippocampus
cerebral peduncle
lateral ventricle
III.
III.
lateral ventricle fimbria
lateral ventricle
habenula
thalamus
III.
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hippocampus
medial geniculate body aquaeduct
red nucleus
lateral ventricle
substantia nigra cerebral peduncle
rostral colliculus
pons
cerebral peduncle
aqueduct
caudal colliculus
midbrain
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cerebellar cortex cerebellar white matter
cerebellar nuclei
fourth ventricle
pyramid
lateral recess
cranial nerve nuclei reticular formation
caudal cerebellar peduncle
trigeminal spinal tract
pyramid
Cranial nerve nuclei
brainstem, cerebellum
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Formalin fixation
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Macroscopical examination of brain and spinal cord
Fixed – Whole organ – Transverse sections
Examination of all structures and anatomical regions – Meninges – Vessels – Cranial nerves/Nerve roots – Hypophysis – Grey/white matter – Ventricular system – Choroid plexus
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Characterisation of lesions
Alterations in Size/Shape/Symmetry
Space occupying (mass) /lytic (malacia) lesion
Color/Consistency
Demarcation
Distribution
– Focal/multifocal/diffuse
– Unilateral/bilateral - symmetrical/asymmetrical/random
– Structures affected
Grey/White matter, specific anatomical structures?
Foreign material: E.g. gun pellets, parasites
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ESAVS 2014
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EASVS 2007
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Macroscopical examination
Transversal sections
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Sampling for histology
Directed by – gross findings
– neurological examination
– diagnostic imaging
Representative sampling
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Representative sampling
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Stereotactic Biopsy
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General strategy
Macroscopical examination Histological examination
Anatomical distribution
Disease category
Clinical history
Literature
VITAMIN D!
Lesion patterns!
Further examinations
Neuroanatomical localisation? Diagnostic imgaging?
eg special stains, immunohistochemistry, bacteriology, TEM, PCR
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Histochemical stainings
Fundamental: H&E
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Grey matter – H&E
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White matter – H&E
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Stainings for axons
Mod. Bielschowsky Holmes
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Stainings for Nissl substance/neurons
Cresyl-Echt-Violet (+ Luxol fast blue: Klüver-Barrera)
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Stainings for myelin
Marchi’s stain Luxol-Fast Blue/H&E
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Stainings for infectious agents
Grocott methamine silver
PAS
Mucicarmin
Gram
Ziehl-Neelsen/Fite
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Immunohistochemistry
In situ method for protein detection
Ramos-Vara, Vet Pathol 2005
ABC-method LSAB-method
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Cellular markers
e.g.
Astrocytes: GFAP
Oligodendrocytes: Olig-2, CNP
Microglia: IBA-1, CD68
Neurons: Synaptophysin, NeuN, Neurofilaments, PGP9.5, …
GFAP
CNP Synaptophysin
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Detection of infectious agents
Listeria monocytogenes Canine distemper
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Artefacts
- White and grey matter vacuolation - Sloughing of ependymal cells
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“Dark neurons”
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Fading of granule cells
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cc
lv
III
Pseudolesions
Subpial germinal cell layer Circumventricular organs Primitive precursor cells