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Nerve

Staining

QuestionAnswer
CNS Central nervous system; brain & spinal cord
PNS Peripheral nervous system
Somatic nervous system is: Voluntary
Autonomic nervous system is: Involuntary
Gray matter contains: Mostly nerve cell bodies
White matter contains: Mostly nerve fibers
What are the 3 groups of nervous system tissues? Neuronal cell bodies & processes, glial cells & processes, myelin sheath
Neuron is made up of: A cell body (perikaryon) w/ nucleus, axons & dendrites; large cells 4-135μm
The nucleus of a neuron contains: Predominantly euchromatin & large nucleolus
Neuroglia is: supporting network for CNS
4 types of glial cells: Oligodendroglia, astroglia, microglia, ependymal cells, Mallory PTAH, Holzer
Oligodendroglia are: small, produce myelin sheath, most numerous type
Astrocytes: Protoplasmic in gray matter, fibrous in white matter, support nerve fiber tracts, exchange fluids, gases, form areas of gliosis, Cajal
Nissl substance: tigroid substance, chromidal substance, basophilic in cytoplasm, aggregates of rough ER, Cresyl echt violets
Chromatolysis is: Loss of Nissl substance due to damage/injury
Microglia are: fixed phagocytic cells in brain & spinal cord, rarely stain for
Ependymal cells are: epithelial cells of ventricles & spinal canal, form BBB
Myelin: white fatty axon sheath, lost in paraffin processing, only neurokeratin remains, formed by oligodendroglia in CNS, formed by Scwann cells in PNS, Weil, LFB
In the event of damage or disease that breaks down the myelin sheath: a simple lipid replacement is formed
Myelin is commonly stained for using what? LFB & iron hematoxylin
Cresyl echt violet (Luna & Vacca) facts To ID neurons, demo chromatolysis, 10% NBF 6-8 μm QC: spinal cord
Cresyl echt violet (Luna) solutions Cresyl echt violet + dH2O + Canada balsam-xylene
Cresyl echt violet (Luna) results Nissl substance, nuclei: blue to purple - background: colorless
Cresyl echt violet (Vacca) solutions Cresyl echt violet pH 2.5 + glacial acetic acid + dH2O + abs. alcohol
Cresyl echt violet (Vacca) results Nissl substance, nuclei: blue-purple - background: colorless
Holmes silver nitrate & Bodian methods purpose Demo nerve fibers, nerve endings, & neurofibrils
Holmes silver nitrate method facts 10% NBF 10-15 μm QC: cerebral cortex
Holmes silver nitrate method solutions AgNO3 w/ boric acid, borate, & pyridine + AgNO3 + gold chloride + sodium thiosulfate + hydroquinone w/ sodium sulfite (red.) + oxalic acid
Holmes silver nitrate method results axons, nerve fibers, neurofibrils: black - other: light gray to blue
Bodian method facts 10% NBF 6-8 μm QC: peripheral nerve or cerebral cortex
Bodian method solutions Protargol S (silver proteinate) + gold chloride + sodium thiosulfate + copper ("destain" connective) + hydroquinone+formaldehyde (red.) + oxalic acid + aniline blue
Bodian method results nerve fibers, nuclei: black - background: light gray or blue
Aniline blue for Bodian solution aniline blue + oxalic acid + phosphomolybdic acid + dH2O
Aqua regia solution HCl + nitric acid
Glassware for Bodian method must first be cleaned with? Aquia regia, extremely hazardous
Bielschowsky-PAS stain, microwave modification of Bielschowsky, & Sevier-Munger modification of Bielschowsky purpose Demo presence of neurofibrillary tangles (disease) & senile plaques of Alzheimer's
Bielschowsky-PAS stain facts 10% NBF 8-10 µm QC: CNS tissue, plaques & tangles if possible
Bielschowsky-PAS stain solutions AgNO3 + ammoniacal silver 20% + gold chloride + sodium thiosulfate + developer + periodic acid + Schiff
Bielschowsky-PAS stain results neurofibrillary tangles, peripheral neurites of neural plaques: dark black - axons: black - amyloid, lipofuscin: magenta
Microwave modification of Bielschowsky facts 10% NBF 8 µm QC: CNS tissue, plaques & tangles if possible
Microwave modification of Bielschowsky solutions AgNO3 + ammoniacal silver + sodium thiosulfate + developer + NH4OH
Microwave modification of Bielschowsky results axons, neurofibrils: brown to black - tangles, plaques: dark brown or black - neuromelanin: black - lipofucsin: brown or black
Bielschowsky developer solution formaldehyde + dH2O + nitric acid + citric acid
Bielschowsky ammoniacal silver solution AgNO3 + NH4OH
Sevier-Munger modification of Bielschowsky facts 10% NBF 6-8 µm QC: CNS tissue
Sevier-Munger modification of Bielschowsky solutions AgNO3 + ammoniacal silver w/ sodium carbonate + sodium thiosulfate + formalin
Sevier-Munger modification of Bielschowsky results nerve endings, neurofibrils, neurofibrillary tangles, peripheral neurites of neuritic plaques: black
Mallory PTAH stain & Holzer method facts Demo glial fibers, gliosis, Holzer better, 10% NBF 6-8 µm QC: cerebral cortex
Mallory PTAH stain solutions PTAH + Lugol Iodine + sodium thiosulfate + potassium permanganate + oxalic acid
Mallory PTAH stain results glial fibers, nuclei, myelin: blue - neurons: salmon
PTAH solution hematoxylin + phosphotungstic acid + dH2O (best aged, ripen immediately w/ potassium permanganate)
Holzer method solutions phosphomolybdic acid-alcohol (mordant) + absolute alcohol-chloroform (differentiate) + crystal violet + potassium bromide + differentiating soln.
Holzer method results glial fibers: blue - background: v. pale blue to colorless
Differentiating solution (Holzer) aniline oil + chloroform + NH4OH
Weil method & all Luxol Fast Blue methods purpose Demo myelin
Weil method, Luxol Fast Blue method, & Luxol Fast Blue-Cresyl Echt violet facts 10% NBF 10-15 µm QC: spinal cord or medulla Regressive staining
Weil method solutions ferric ammonium sulfate (excess mordant) + alcoholic hematoxylin in dH2O + Na borate-K ferricyanide (excess ox.)
Weil method results myelin sheath: blue to blue-black - background: light tan
Luxol Fast Blue method solutions LFB in alcohol w/ acetic acid + Li2CO3 + 70%
Luxol Fast Blue method results myelin: blue - background: colorless
Luxol Fast Blue-Cresyl Echt violet solutions LFB in alcohol w/ acetic acid + Li2Co3 + 70% + Cresyl Echt violet
Luxol Fast Blue-Cresyl Echt violet results myelin: blue - Nissl substance, nuclei: violet
Luxol Fast Blue-Holmes silver nitrate method facts 10% NBF 10-15 µm QC: cerebral cortex
Luxol Fast Blue-Holmes silver nitrate method solutions LFB in alcohol w/ acetic acid + Li2CO3 + AgNO3 w/ boric acid, borate, & pyridine + AgNO3 + gold chloride + sodium thiosulfate + hydroquinone w/ sodium sulfite (red.) + oxalic acid
Luxol Fast Blue-Holmes silver nitrate method results myelin sheaths: blue to green - axons, nerve fibers: black
Luxol Fast Blue-PAS-hematoxylin is used to: Demo myelin & basement membrane
Cajal stain facts Demo astrocytes, mostly replaced by IHC, formalin ammonium bromide, if fixed in NBF, wash & post-fix w/ formalin ammonium bromide frozens 20-30 µm QC: cerebral cortex
Cajal stain solutions gold chloride w/ mercuric chloride (gold sublimate) + sodium thiosulfate
Cajal stain results astrocytes: black background: pink
Wallerian degeneration If axon sustains injury, axon & myelin below injury site are lost
Thioflavin S facts To ID neurofibrillary degenerations 10-20% NBF 6 μm QC: CNS w/ senile plaques
Thioflavin S solutions potassium permanganate (quench autofluorescence) + potassium metabisulfite (quench autofluorescence) + H2O2 (quench autofluorescence) + oxalic acid (remove potassium permanganate) + acetic acid + thioflavin S
Thioflavin S results Alzheimer tangles, plaques, neuropil threads, senile plaque & cerebrovascular amyloid: bright green - diffuse plaques & extracellular tangles: pale yellow green
Nerve fibers, nerve endings, neurofibril stains Bodian, Holmes
Neurofibrillary tangles & plaques stains Bielschowsky-PAS, microwave Bielschowsky, Sevier-Munger Bielschowsky
Holzer crystal violet solution crystal violet + abs. alcohol + chloroform
How to remove crystal violet precipitate? Straight aniline oil
Argyrophilic reaction is one where: Tissue can bind silver ions, but need reducing agent to reduce to visible silver
Argentaffin reaction is one where: Tissue can bind silver ions & reduce to visible silver
Purkinje cells: aka Purkinje neurons, cerebellar cortex, flask-shaped w/ many dendrites
PTAH stains tissue components: Red-brown/salmon, components lose color if prolonged water/alcohol washing, dehydrate rapidly after staining
Page's Eriochrome Cyanine R purpose Demo myelin
Created by: CCF
 

 



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