Nucleic Acid Stains
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| Two types DNA AND RNA | Nuclei Acid
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| Has 5 carbon sugars called Deoxyribose, Found in nucleus, its major constituent is nuclear chromatin | DNA
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| Has 5 carbon sugars called Ribose, found in the nucleolus and ribosomes | RNA
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| The sugars in DNA and RNA differ by a single hydroxyl group (OH-). the difference allows for selective demonstration of DNA and RNA by special staining technique | DNA AND RNA
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| process by which a chemical compound decomposes by reacting with water | Hydrolysis
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| demonstrates DNA, hydrolysis of DNA by hydrochloric acid | Feulgen stain
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| With Feulgen DNA is hydrolyzed by 1N HCl which removes the purine (adenine and guanine)base but leaves the sugars and phosphates of DNA intact, this generates an aldehyde which can be demonstrated with Schiff's reagent | Feulgen stain
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| time of hydrolysis must be carefully controled, after removal of purine bases and formation of aldehyde groups, there is a progressive removal of histones and apurinic acids, second reaction leads to false negative feulgen reaction | Feulgen stain
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| DNA appreas Magenta/reddish purple | Feulgen stain
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| cytoplasm stains only if a counter stain is applied (light green) | Feulgen stain
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| Dont use BOUINS to fix tissues it hydrolyzes nuclei excessively, dont overcounterstain with light green | Feulgen stain special consideration
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| masked feulgen | overcounterstained with light green
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| NONE required all nuclei will stain | Feulgen control tissue
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| purpose to differentiate between RNA and DNA | Methyl green pyronin Y
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| differential staining is due to differing degrees of polymerization between DNA and RNA | Methyl green pyronin Y
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| DNA high Polymer, RNA lower Polymer | Methyl green pyronin Y
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| used to identify plasma cells and immunoblasts in tissues | Methyl green pyronin Y
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| DNA binds w methyl green (green/blue color), RNA binds with Pyronin (red color), background pale pink to colorless, mucins, epilthelium and cartilage appear pink to red | Methyl green pyronin Y
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| Carnoyed fixed best, 10%NBF works, B-5, Helly or Zenker, | Methyl green pyronin Y fixatives
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| Control tissue, a section containing numerous plasma cells | Methyl green pyronin Y Controls
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| acids and fixatives may depolymerize DNA resulting in loss of methyl green staining, While the RNA bind to pyronin. resulting in loss of differential staining. | Methyl green pyronin Y special consideration
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| pyronin is not specific for RNA. cartilage, osteoid, keratin, eosinphil granules and mast cell granules will also stain, DNA will stain but pyronin cant compete with Methyl green for DNA | Pyronin
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| process where a dye forms other dyes spontaneously | Polychroming
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| A compound dye or dye mix that contains components of different colors | Polychromatic stain example
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| Most common example of Romanowsky stain | May-Grunwald Giemsa
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| stains for hematopoietic tissues another group of nuclear and cytoplasmic special staining technique | May-Grunwald Giemsa
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| where red and white blood cells are formed | hematopoietic tissues
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| stains used to identify different cell lineages found in hematopoietic tissues, like spleen,bone marrow and blood | May-Grunwald Giemsa
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| basophilic or basic dye, methylene blue is combined w/eosinphilic or acid dye, to create "neutral dyes" that demonstrate a wide variety of colors when used to stain hematopoietic cell nuclei and platelets. | May-Grunwald Giemsa principle (chemistry)
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| differentiate cells present in hematopoietic tissue. demonstrates presence of microorganisms. hitological dx for malaria, spirochete and protozoan blood parasites | May-Grunwald Giemsa
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| Nuclei stains blue, cytoplasm leukocytes pink, gray or blue, bacteria stains blue | May-Grunwald Giemsa
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| Zenker or B-5 preferred, 10%NBF ok to use | May-Grunwald Giemsa fixative
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| working solutions not stable, prepare solutions before use and then discard. | May-Grunwald Giemsa
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| if staining is poor, the effect of the pH adjustment should be investigated. pH should be between 6.4-6.9 | May-Grunwald Giemsa
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| Control spleen | May-Grunwald Giemsa
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| made of 1-2% carbohydrate (chondroitin,heparin,and dermatan)and acid mucopolysaccharides | Amyloid
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| disease charactarized by amorphorous, eosinphilic, extracellular deposit, that slowly replace cellular elements of vital organs and causes progressive loss of function and eventually death | amyloidosis
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| varies between patients and between organs in the same patient | composition of amyloid
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| occurs spontaneous in the absence of any predisposing disease | primary amyloidosis
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| organs affected are muscle, heart, skin, tongue | primary amyloidosis
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| associated with a predisposing disease, chronic inflamatory disease rheumatoid arthritis, ankylosing spondylitis, infections such as tuberculosis and osteomyelitis | secondary amyloidosis
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| this type of amyloid most frequently deposited in kidneys, liver, spleen and adrenal gland | secondary amyloidosis
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| associated with disease of the immunological system, resembles primary amyloid | Myeloma-associated amyloid
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| found w/many tumors APUD | Tumor associated amyloid
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| group of cells of embryonic origin that secrete most of the bodys hormones | APUD celss
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| compromise both specialized neurons and endocrine cells.synthesize structually related polypeptides and biogenic amines | APUD cells
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| amine precursor uptake and decarboxylation | APUD
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| insulin, ACTH, glucagon,antidiuretic hormes | peptide hormone examples
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| dopamine, norepinephrine, serotonin and histamine | amine hormone examples
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| oncology tumor that produces small peptide hormones of the APUD system | Apudoma
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| small-oat cell carcinoma of lung, carcinoids of lung, thymus, GI tract and prostate, medullary CA of thyroid, pancreatic islet cell tumor, malignant melanoma, Ganglioneuroma | Apudoma Examples
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| linked to uptake of precursor amino acid and its decarboxyaltion in the cell to produce an amine | polypeptide production
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| immunoglobulin derived and includes primary amyloid and myeoloma associated amyloid | AL amyloid
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| Unknown origin and includes secondary amyloid | AA amyloid
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| heredofamilial amyloid (familial mediterranean fever) | AF amyloid
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| polypeptide-hormone derived,associated with tumors of the APUD system | APUD
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| demonstrates amyloid in tissues, most specific technique to demonstrate amyloid, | Alkaline congo red
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| pretreatment with alkali aids in the release of native internal hydrogen bonds between adjacent protein chains creating mores sites for the dye to bind | alkaline congo red
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| amyloid stains deep pink to red with light microscope | congo red
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| amyloid stains bright green apple with polorazing microscope | Congo red
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| elastic tissues stain pale pink | congo red
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| nuclei stains blue | congo red
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| cut section at 8-10 microns | congo red
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| any tissue w/amyloid, staining intensity decreased w/time, dont cut too many control slides ahead of time | congo red special considerations
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| need polorazing scope to view green apple birefringence | congo red
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| rapid screening method to demonstrate amyloid | crystal violet
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| not as specific as congo red | crystal violet
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| addition of HCl in the stain prevents the cytoplasmic component from overstatining | crystal violet
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| amyloid stains violet, other tissue elements stain blue | crystal violet
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| cut sections 10-12 microns | crystal violet
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| any tissue with amyloid | crystal violet control
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| cover slip w/apathy mounting media or aqueous based mounting media or air dry slide completely then dip in xylene and use resinous media | crystal violet
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| purpose demonstrate presence of amyloid. not as good as congo red w/polarizing light, background nuclear fluoresece is quench w/ aluminum hematoxylin. no differentiation is required | Thioflavin T fluorescent Method
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| Amyloid fluoresces yellow/yellow green | Thioflavin T fluorescent Method
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| Sections at 6-10 microns | Thioflavin T fluorescent Method
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| Thioflavin T at acidic concentration increases the selectivity of the dye for amyloid, pH 1.4 is recommended | Thioflavin T fluorescent Method special considerations
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| lipid granules, mast cells and juxtaglomerular granules may give yellow fluoresence | Thioflavin T fluorescent Method
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| pH of staining solution critical. More acid levels give more selective chromatin staining and less cytobasophilia, less acid pH levels give denser nuclei and increased cytoplasmic basophilia | May-Grunwald Giemsa
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