Nuclear and cytoplasmic staining in the histology lab.
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show | Nucleus and Cytoplasm
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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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show | Nuclear pores
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show | Nucleolous
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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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Consists of chromosomes or DNA and attached protein | show 🗑
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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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show | Ribosomes
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show | Ribosomes
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show | Golgi apparatus
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show | Centriole
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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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Long lived or permanent cells such as neurons, cardiac muscle, and hepatocyte accumulate a large amount of residual bodies. it is referred to as | show 🗑
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Exogenous materials | show 🗑
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Melanin and hemoglobin breakdown | show 🗑
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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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Ionic bonding | show 🗑
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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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Covalent bonding | show 🗑
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Elements that typically form covalent bonds | show 🗑
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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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show | proteins or charged groups on the side chains of amino acids constituting the proteins
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Proteins that can be positive or negative | show 🗑
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Eosin must be kept below pH of 6 or else | show 🗑
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A group that confers the property of color | show 🗑
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if reduction occurs to a chromopohore what happen? | show 🗑
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show | Chromogen
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show | Auxochrome
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Amino (NH2) | show 🗑
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Chromophore | show 🗑
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Picric acid is | show 🗑
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show | Sulfonic, carboxyl, and hydroxyl groups
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Cationic dyes | show 🗑
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show | Crystal violet and safranin
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show | Orange G and picric acid
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show | Hematein and lithium carminate
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Factors that affect dye binding | show 🗑
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show | Formalin
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show | Potassium dichromate
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Tissue will lose its nuclear staining properties when stained with these types of fixatives | show 🗑
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Types of fixatives that increase tissue basophilia or the uptake of cationic or positively charged dyes | show 🗑
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show | Picric acid
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show | Mordants
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Ways to differentiate | show 🗑
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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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Hematoxylin can be oxidized by | show 🗑
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show | Hematoxylin
Absolute Alcohol
Ammounium aluminum sulfate
Distilled water
Mercuric Oxide
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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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show |
Solution A
Ammonium aluminum sulfate
Distilled water
Solution B
Hematoxylin
95% alcohol
Glycerol
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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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To check for over oxidation of Delafield heme | show 🗑
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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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Mayer heme mordant and oxidizer | show 🗑
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Citric Acid and Chloral hydrate do what in the Mayer heme solution? | show 🗑
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Recommended heme for immunoperoxidase techniques when 3-amino-9-ethylcarbazole is used | show 🗑
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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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Ehrlich heme mordant and oxidizer | show 🗑
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show | More commonly used regressively but can be used progressively
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Gill heme | show 🗑
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Ethylene glycol does what in the Gill heme? | show 🗑
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show | Mordant: Aluminum sulfate
Oxidizer: Sodium iodate
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Gill 2 and 3 are used for | show 🗑
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Gill 3 is | show 🗑
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Can be used for staining glycol methacrylate sections | show 🗑
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Gill 1 and 2 are used | show 🗑
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show | Goblet cells in Mucin
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Less prone to surface precipitate | show 🗑
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Scott Solution | show 🗑
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show | Changes the pH and allows the slides to blue (change color) Also stabilizes the stain.
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show | Lithium carbonate, ammonium hydroxide, or Scott solution
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show | Ferric chloride
Distilled water
hydrochloric acid
Hematoxylin
95% alcohol
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show | Weigert heme
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show | Mordant and oxidizer: Ferric Chloride
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Substitute for Weigert heme | show 🗑
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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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show | Too much heat during processing or drying of the microscopic sections
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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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Pale nuclear staining can be corrected by | show 🗑
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show | 1. Sections left too long in heme
2. Sections too thick
3. Differentiation step too short
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Dark nuclear staining can be corrected or prevented by | show 🗑
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show | Either the heme is breaking down or blueing step was not properly done
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show | -Ensure sections are blued properly
-Check the oxidation of the heme
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Pale cytoplasmic staining may result from | show 🗑
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Pale cytoplasmic staing can be prevented or corrected from | show 🗑
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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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Eosin not properly differentiated may be prevented or corrected by | show 🗑
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Blue-black precipitate on top of sections | show 🗑
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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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show | Laser and electrocautery techniques (No remedy)
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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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DNA is found in the | show 🗑
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RNA is found in the | show 🗑
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Feulgen reaction purpose | show 🗑
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Feuglen reaction principle | show 🗑
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show | Anything but Bouins
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show | DNA: Reddish purple
Cytoplasm: Light green
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Methyl green-pyronin Y purpose | show 🗑
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show | Methyl green-pyronin Y
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Methyl gree-pyronin Y principle | show 🗑
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show | 10% NBF is preferred. B-5, Helly, or Zenker is fine
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Methyl green pyronin Y results | show 🗑
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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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May Grunwald Giemsa stain Purpose | show 🗑
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May Grunwald Giemsa stain Principle | show 🗑
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show | Zenker or B-5 preferred, 10% NBF is fine
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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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Tissue fixed in aldehydes will have | show 🗑
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show | Resinous and Aqueous
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Resinous media tends to be | show 🗑
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Examples of resinous media | show 🗑
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show | Xylene
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Natural resins | show 🗑
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show | Toulene
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Tissues average refractive index | show 🗑
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Synthetic resins refractive index | show 🗑
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show | Mineral oil
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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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show | Dehydrating and clearing will adversely affect the stain
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Aqueous mounting media refractive index | show 🗑
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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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