Nuclear and cytoplasmic staining in the histology lab.
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| Two majors parts of a cell | show 🗑
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| Resting nucleus is typically in what phase? | show 🗑
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| The nucleus appears to be what color when being stained with the standard H&E stain? | show 🗑
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| Electron microscopes are able to view subcellular particles which include | show 🗑
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| Most likely the avenues of communication between the cytoplasm and the nucleus | show 🗑
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| Usually intensely basophilic but with a good H&E stain it is acidophilic | show 🗑
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| Produces most of the ribosomal RNA | show 🗑
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| show | Stainable
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| Euchromatin | show 🗑
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| show | Chromatin
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| show | Heterochromatin
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| show | Lymphocytes
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| show | Neuronal nuclei
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| Energy producing powerhouses of the cell | show 🗑
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| Site of protein synthesis | show 🗑
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| Some are on the endoplasmic reticulum while others are located in the cytoplasm | show 🗑
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| show | Ribosomes
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| show | Golgi apparatus
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| Responsible for spindle formation in cell division | show 🗑
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| show | Centrioles
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| show | Lysosomes
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| Aid in the digestion of food taken into the cell | show 🗑
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| show | Lipofuscin (wear and tear pigment)
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| show | Carotene, dusts, and minerals
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| show | endogenous pigments
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| Attraction for minute particles from the surrounding solution, by the surface of certain tissue components; the dye is then bound to the tissue primarily by ionic, covalent, or hydrogen bonds | show 🗑
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| show | Different charges that become attracted to one another
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| show | Covalently bonded hydrogen is attracted to atoms that have a strong electronegative charge.
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| Hydrogen frequently bonds to what elements | show 🗑
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| show | When atoms share electrons
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| show | Carbon, hydrogen, and oxygen
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| show | Caused by electrostatic attraction of a molecule for the electrons of its neighboring molecules. These are weak physical forces that are effective over only very short distances.
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| show | 1. Done with basic (cationic or positively charged) dyes
2. Done with dyes combined with or followed by metal mordants
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| show | Presence of the nucleic acids (DNA and RNA) to form dye salt type unions
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| show | Nucleic acids have been removed (decalcified tissue) and also may occur in tissue that is not negatively charged.
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| Non nuclear staining is primarily cause by | show 🗑
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| Proteins that can be positive or negative | show 🗑
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| show | the -COO group of eosin recombines with hydrogen and the result is the free acid, uncharged form of eosin. Non specific staining will occur.
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| A group that confers the property of color | show 🗑
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| show | Color is lost
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| A benzene derivative containing chromophoreic groups | show 🗑
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| An ionizing group that links firmly to the tissue | show 🗑
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| show | Auxochrome
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| show | C=O, C=S, C=N, N=N, N=O,NO2
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| Picric acid is | show 🗑
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| Anionic auxochromes | show 🗑
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| Cationic dyes | show 🗑
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| Basic dyes | show 🗑
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| Acid dyes | show 🗑
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| Amphoteric dyes | show 🗑
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| Factors that affect dye binding | show 🗑
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| show | Formalin
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| Tissue binds less with Heme when this type of fixative is used | show 🗑
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| show | Zenker, Bouin, unbuffered formalin, (all acidic fixatives)
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| show | Formaldehyde, mercuric chloride, and osmium tetroxide
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| show | Picric acid
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| show | Mordants
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| show | 1. When basic or cationic dyes are used differentiate by weak acid
2. Excess mordant
3. oxidizers
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| show | Oxidizers
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| show | Iron heme stains
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| Aluminum hemes can be differentiated with | show 🗑
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| Eosin can be differentiated with | show 🗑
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| Hematein | show 🗑
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| show | air, light, sodium iodate, mercuric oxide, and potassium permanganate
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| Harris Heme | show 🗑
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| show | Sodium Iodate
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| Harris Hematoxylin mordant and oxidizer | show 🗑
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| show | Progressively and acidified
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| Delafield hematoxylin | show 🗑
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| show | Mordant: Aluminum
Oxidizer: light and air
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| show | Stabilizes the solution against over oxidation and prevents rapid evaporation
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| show | Drop a few drops into a container of water. If it is blue-black it can still be used. If the solution turns into a red or red brown then it will be over oxidized.
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| How to check for under oxidation of Delafield heme | show 🗑
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| Delafield heme is used | show 🗑
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| Mayer heme | show 🗑
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| show | Mordant: Aluminum
Oxidizer: Sodium Iodate
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| Citric Acid and Chloral hydrate do what in the Mayer heme solution? | show 🗑
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| show | Mayer heme is used because it does not contain alcohol and will not dissolve the reaction product.
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| Very difficult to over stain with this heme and produces very crisp nuclear staining. (Progressive) | show 🗑
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| show | Hematoxylin
Alcohol 95%
Distilled water
Glycerol
Ammonium or potassium aluminum sulfate
Glacial acetic acid
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| show | Mordant: Aluminum
Oxidizer: natural or chemical can be used
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| Ehrlich heme | show 🗑
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| show | Distilled water
Ethylene glycol
hematoxylin, anhydrous
Sodium Iodate
Aluminum sulfate
Glacial acetic acid
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| show | Prevents the formation of surface precipitate
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| Mordant and oxidizer of Gill heme | show 🗑
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| Gill 2 and 3 are used for | show 🗑
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| show | most concentrated
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| Can be used for staining glycol methacrylate sections | show 🗑
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| show | Progressively
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| show | Goblet cells in Mucin
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| show | Mayer and Gill
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| Scott Solution | show 🗑
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| Scott solution | show 🗑
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| Weakly alkaline solutions | show 🗑
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| Weigert heme | show 🗑
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| show | Weigert heme
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| Weigert mordant and oxidizer | show 🗑
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| show | Gallein iron heme
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| how many shade of pink should be obstained when staining with Eosin | show 🗑
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| The more dilute alcohol the more _ will be removed | show 🗑
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| show | the A
The longer it may need to stain
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| show | Helly, Zenker, or B-5
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| show | Place in 5% aqueous lithium carbonate for 1 hour
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| Restoring tissue basophilia: If over exposed to Zenker | show 🗑
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| Restoring tissue basophilia: Method III | show 🗑
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| Tissue basophilia loss may result from | show 🗑
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| show | Weigert iron heme
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| show | incomplete deparaffinization
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| show | -Ensuring the sections are dried properly
-Allow enough time in xylene
-Ensure xylene is not contaminated
-If slides are stained then decolorize and restain
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| When distinct chromatin pattern cant be seen (Smudgy or muddy nuclear detail) | show 🗑
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| Most frequent cause of nuclear staining not being crisp | show 🗑
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| Other causes of nuclear staining not being crisp | show 🗑
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| show | -Complete fixation of specimens
-Tissues should be dehydrated and cleared completely
-No heat on processor
-Tissue should not remain in melted paraffin for long
-Dryer is correct temp
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| Pale nuclear staining | show 🗑
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| Over decalcified specimen | show 🗑
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| show | -Ensuring slides are left in heme long enough
-Ensure heme is not overoxidized
-Ensuring differentiation step is properly times
-restaining section
-use of fixative
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| Dark nuclear staining most likely causes | show 🗑
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| show | -Ensuring sections are thing
-Decolorize the sections and restain
-Decrease the time sections remain in heme
-Increasing the time of differentiation
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| show | Either the heme is breaking down or blueing step was not properly done
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| Ways to prevent or correct red/red-brown nuclei | show 🗑
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| Pale cytoplasmic staining may result from | show 🗑
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| show | -Checking the pH of Eosin
-Ensure the blueing reagent is removed before transferring the slides to eosin
-Ensure that stained slides are not allowed to stand in low concentrations of alcohols after eosin
-Ensure sections are not too thin
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| show | -Ensure the eosin solution is not too concentrated
-Ensure sections are not left too long in Eosin
-Ensure time in dehydration solutions allow good differentiation
-Check section for proper thickness
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| show | -Ensuring timely and complete fixation
-Ensure good dehydration and clearing during processing
-Ensure eosin stained sections get proper differentiation
-Ensure eosin is correct pH
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| show | Caused from the metallic sheen that is formed on top of heme solutions (Filter to correct)
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| show | if after rehydrating alcohols then it indicates the presence of xylene. Back the slides up and change the alcohols. The take slides from alcohol to water. Water should be clear
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| show | Water is still present. Back up the slides and change the alcohol and xylene. Then rehydrate and clear sections.
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| show | Caused by water or fixative infiltrating paraffin caused by contamination of reagents in closed tissue processors because of equipment malfunction or absorbtion of atmospheric water by dehydrating alcohols on the open processors
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| show | -Change from xylene to toluene in areas of high humidity if using an open processor
-Have equipment checked for malfunctions
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| Dark basophilic staining of nuclei and cytoplasm (Especially around the edges | show 🗑
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| Poor contrast between nuclei and cytoplasm causes | show 🗑
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| Poor contrast between the nuclei and the cytoplasm can be prevented or corrected by: | show 🗑
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| show | nucleus
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| show | nucleolous and ribosomes
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| show | Demonstration of DNA
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| Feuglen reaction principle | show 🗑
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| Feuglen reaction fixative | show 🗑
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| Feuglen reaction results | show 🗑
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| Methyl green-pyronin Y purpose | show 🗑
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| Primarily used to identify plasma cells and immunoblasts in tissue | show 🗑
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| show | DNA stains with green, while RNA is colored red with pyronin. Differential staining caused from differing degrees of polymerization. Methyl green is bound by more highly polymerized DNA.
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| Methyl green pyronin Y fixative | show 🗑
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| show | DNA: Green to blue-green
RNA: rose
Goblet cells: Mint green
Background: Pale pink to colorless
Immunoblast and plasma cell cytoplasm: Intense red
Nuclei: Green to blue-green
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| show | A compound dye or dye mixture that contains components of different colors
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| A process in which a dye forms other dyes spontaneously | show 🗑
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| Romanowsky type stain | show 🗑
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| show | To permit differentiation of cells present in hematopoietic tissue. Also used to stain certain microorganisms
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| May Grunwald Giemsa stain Principle | show 🗑
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| May Grunwald Giemsa stain Fixative | show 🗑
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| show | Spleen
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| show | Nuclei: Blue
Cytoplasm: Shades of pink, gray, blue
Bacteria: Blue
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| show | increased basophilia
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| show | Resinous and Aqueous
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| show | natural
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| show | Canada balsam and gum dammar
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| Natural resins dissolve in what? | show 🗑
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| Natural resins | show 🗑
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| Most resinous media are dissolved in | show 🗑
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| show | 1.53-1.54
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| show | 1.51-1.55
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| Best preservative to protect Romanowsky stain when using a natural resin | show 🗑
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| show | Remove coverslip and any remaining medium with xylene. Rehydrate with the appropriate reagent, clear with xylene, and remount with synthetic resin
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| Aqueous mounting media is used when | show 🗑
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| show | 1.41-1.43
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| show | 1 1/2 (180 micrometers thick), number 1 coverslips (~150 micrometers thick)
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| show | Section transparency is reduced
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| Thickening of the mounting medium does what to transparency? | show 🗑
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| show | Results from thickened medium or excess medium between the section and cover glass
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| If drying artifact is noted what should be done to correct it? | show 🗑
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| Exhibits brown stippling that resembles pigment or nucleus appears ti have a glossy black structure | show 🗑
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