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140 Final HTO
FINAL EXAM PREP!!!!!!!
| Question | Answer |
|---|---|
| H & E Trouble shooting: Produced from heat artifact, laser techniques | Dark basophilic staining around edges of tissue |
| H & E Trouble shooting: Left too long in hematoxylin | Dark nuclear staining |
| H & E Trouble shooting: Not enough time in hematoxylin | Pale nuclear staining |
| H & E Trouble shooting: Due to incomplete fixation on the processor | Nuclear staining not crisp |
| H & E Trouble shooting: Not enough adequate time in alcohol for proper differentiation | Eosin not properly differentiated |
| H & E Trouble shooting: Unfiltered hematoxylin | Blue-black precipitate on top of sections |
| H & E Trouble shooting: Incomplete drying, not enough time in xylene and water left in tissue | Incomplete deparaffinzation |
| H & E Trouble shooting: Section left in eosin too long | Dark Cytoplasmic staining |
| H & E Trouble shooting: Hematoxylin is breaking down | Red or Red-Brown Nuclei |
| H & E Trouble shooting: High eosin pH/sections too thin | Pale Cytoplasmic staining |
| What is the only saccharide that can be demonstrated histologically? | Polysaccharide |
| What stain will demonstrate only sulfated mucosubstances? | Alcian Blue pH 0.4- 1.0 |
| What oxidizer is used in Bauer Feulgen? | Chromic Acid |
| Neutral mucosubstances are demonstrated by what special stain? | PAS |
| What enzyme is commonly used to breakdown glycogen? | Diastase |
| What stain is commonly used to demonstrate acid mucoploysaccharides? | Alcian Blue |
| Which stain is used for metachromatic dyes in connective tissue? | Toluidine Blue |
| List two control tissues used for mucicarmine | GI, Small Intestine |
| Is Mayer's Mucicarmine a Synthetic dye or natural dye? | Natural |
| Name the two specific mordants and oxidizers used in elastic stain? | 10% ferric chloride and Lugol Iodine |
| What reagent is used to remove excess iodine in the elastic stain? | Sodium Thiosulfate |
| Wat reagent is used as the differentiator for elastic stain? | 2% ferric chloride |
| What type of stain is Verhoeff's? | Regressive stain |
| What fixative is used for Masson Trichrome? | Bouin's |
| What is the blue dye is tissue element called in a Trichrome stain? | Collagen |
| What dye is used to stain the tissue element blue in the trichrom stain | Collagen |
| What dye is used to stain the tissue element blue in the Trichrome stain? | Aniline Blue |
| What is the red dye tissue element called in a Trichrome stain? | Cytoplasm/Muscle |
| What dye is used to stain the tissue element red in a Trichrome stain? | Biebrich Scarlet Acid Fuchsin Solution |
| What reagent is used to remove excess Potassium Permanganate in the Retic stain? | Potassium Metabisulfite |
| What is the primary fixative used in the PTAH stain? | Zenker's or Helly |
| What are the blue tissue elements shown in the PTAH stain known as? | Skeletal |
| What is the purpose of Ferric Ammonium Sulfate in the silver procedure? | Sensitization |
| What is the purpose of Gold Chloride in the silver stain procedure? | Toning |
| What counterstain is used in Alcian Blue stain? | Nuclear Fast Red |
| What control tissue is preferred for Alcian Blue stain? | Appendix |
| What is primary fixative for PTAH stain? | Zenker |
| what is the preferred thickness of PTAH stain? | 4-6 microns |
| What reagent is used as the oxidizer in PTAH stain? | Potassium Permanganate |
| What reagent is used as the decolorizer in PTAH stain? | Oxalic Acid |
| What reagent is used as the oxidizer in PAS stain? | Periodic Acid |
| What reagent is used to stain the tissue elements bright pink to rose for PAS stain? | Schiff's Reagent |
| What is the purpose of PAS stain? | Identify neutral mucosubstances |
| Besides 10% NBF, what is the preferred fixative for PAS stain? | Absolute Alcohol |
| Besides 10% NBF, what is the preferred fixative for crystal violet? | Absolute Alcohol |
| What is the preferred thickness for crystal violet? | 10-12 microns |
| What type of control tissue is preferred for crystal violet? | amyloid |
| What type of mounting media is used for crystal violet? | aqueous |
| What reagent is used as the sensitizer in Alcian Blue/PAS? | periodic acid |
| What is the counterstain used in Alcian Blue/PAS? | hematoxylin |
| What are the blue tissue elements known as in Alcian Blue/ PAS? | acid mucosubstances |
| What reagent is used in gram stain as the mordant? | gram iodine |
| What is used to identify the positive organisms in gram stain? | crystal violet |
| what reagent is used to identify negative organisms in gram stains? | safranin O |
| What reagent is used as a counterstain in gram stain? | tartrazine |
| What reagent is used as the sensitizer in Steiner stain? | uranyl nitrate |
| What reagent is used as the resin in Steiner? | Gum mastic |
| what reagent stains the organisms in Steiner? | silver nitrate |
| What fixative is used for steiner? | 10% NBF |
| What reagent is used to stain the organism in AFB stain? | carbol fuchsin |
| what reagent is used as the counterstain for AFB? | methylene blue |
| what reagent is used to decolorize AFB stain? | acid alcohol |
| What reagent is used as the oxidizer for GMS-F stain? | chromic acid |
| what reagent is used to remove the oxidizer in the stain? | sodium bisulfite |
| microscopic evaluation of an H&E stained section reveals dark deposits of material dispersed irregularly over the tissue. the most likely cause of this problem is that the hematoxylin solution: _______ | was not filtered before use |
| Following h&E staining, slides are dehydrated through ascending strengths of alcohols and cleared in xylene; however, the first xylene in the series is milky white. the most appropriated action is to: ________ | change alcohols and xylenes at the end of the series |
| tissue sections were stained for the recommended time with an h&E procedure using Harris Hematoxylin. A quality control check shows pale nuclear staining. A likely cause of this could be: _________ | too much time in the differentiating solution |
| microscopic examination of an H&E stained section shows pink artifact surrounding the tissue and in tissue spaces. the most probable cause of this artifact is: _________ | precipitation of the eosin |
| What reagent is added to the staining line to create the regressive H &E staining method? | acid alcohol |
| What reagent is added to Harris Hematoxylin to make it a working solution? | Glacial acetic acid |
| A blue-black precipitate is seen on a h&E stained sections. this could probably be prevented in the future by: _____ | filtering the hematoxylin |
| What is the counterstain used for Retic stain? | Nuclear fast red |
| What two reagents are used to stain the tissue element black for Retic stain? | Silver nitrate & ammonium Hydroxide |
| What is the oxidizer used in Retic stain? | Potassium permanganate |
| What is the sensitizer used in Retic stain? | Ferric Ammonium Sulfate |
| What reagent is used to reduce the background staining for congo red? | sodium chloride |
| Besides 10% NBF, what fixative can be used for congo red? | carnoy |
| what is the preferred thickness for congo red? | 8-10 microns |
| The pink to red deposits are called ________ in Congo red? | Amyloid |
| what reagents are used to act as the mordant and oxidizer for EVG stain? | Lugol iodine & 10% ferric chloride |
| Besides 10% NBF, what fixative can be used for EVG stain? | Zenker |
| What control tissue is preferred for EVG? | Artery |
| What two reagents are used to create the counterstain for EVG? | Acid Fuchsin & Picric Acid |
| What reagent stains the blue tissue elements in trichrome? | Aniline Blue |
| What are the blue tissue elements known as for trichrome stain? | collagen |
| What reagent stains the red tissue elements in trichrome stain? | Biebrich Scarlet Acid Fuchsin |
| What mordant is used in Trichrome stain? | Bouin |
| What control tissue is preferred for mucicarmine? | small intestine |
| what reagent is staining the tissue elements deep rose to red for mucicarmine stain? | mucicarmine |
| what counterstain is used for mucicarmine? | metanil yellow |
| The tissue elements being stained deep rose to red are known as goblet cells | goblet cells |
| Besides 10% NBF, what is the preferred fixative for PAS/D stain? | Bouin |
| What reagent is used for digestion for PAS/D? | Diastase |
| What control tissue can be used for PAS/D? | Liver |
| What is the purpose of PAS/D stain? | Identify neutral mucosubstance. |
| what are the preferred microns for luxol fast blue stain? | 10 - microns |
| besides the spinal cord, what control tissue is needed for the bielschowsky stain? | brain from Alzheimer's patient |
| whareare the preferred microns for the bielschowsky stain? | 8-10 microns |
| what reagents are used as the differentiators in the luxol fast blue stain? | lithium carbonate/ 70% alcohol |
| besides the spinal cord, what control tissue is needed for the luxol fast blue stain? | medulla |
| what type of stain is the bielschowsky? | argyrophilic |
| name the four reagents used in a bielschowsky stain. | FILL FILL FILL!!!!!!!!!!!!!!!!!! |
| what fixative is used for Oil Red O and Sudan Black B? | Formol Calcium |
| What is the preferred thickness for Oil Red O and Sudan Black B stains? | 7-10 microns |
| In the Oil Red O stain, what reagent is used to dissolve dye? | Propylene Glycol |
| what reagent is used as the counterstain in Sudan Black B? | Nuclear Fast Red |
| What type of mounting media is used to coverslip both Oil Red O and Sudan Black B slides? | Aqueous mounting media |
| In the Oil Red O stain, what is the color of the neutral lipids? | Red |
| In sudan black b stain, what is the color of the neutral lipids? | black |
| bothoil red o and sudan black b are what type of staining reaction? | physical |
| explain why it is a good practice to not press on the coverslip of oil red o and Sudan black b slides? | can push the fat out of the tissue |
| name one type of solvent that should be avoided with lipids. | alcohol fixation |
| H&E troubleshooting: White spots seen in tissue sections. What is it called and how do you prevent it? | Incomplete Deparafffinization prevent by: dry sections properly before deparaffinizing, allow for sufficient time in xylene, avoid contaminated xylene. |
| H&E troubleshooting: Smudgy or muddy nuclear staining. What is it called and how do you prevent it? | Nuclear staining is not crisp. prevent by: fix tissue speciemens completely, dehydrate and clear tissues completely before infiltrating with paraffin. do not use heat on processor except for paraffin. dry slides at correct temperature. |
| H&E trouleshooting: Hematoxylin is too light. what is it called and how do you prevent it? | Pale nuclear staining Prevent by: not leaving slides in hematoxylin long enough. staining with overoxidized or depleted hematoxylin. overdiffferentiating the hematoxylin. |
| H&E troubleshoot: Nuclei is too darkly stained. what is it called and how do you prevent it? | Dark nuclear staining Prevent by: sections left too long in hematoxylin. sections too thick differentiation step too short. |
| H&E troubleshoot: Nuclei is stained red or reddish brown instead of blue. what is it called and how do you prevent it? | Red or red-brown nuclei Prevent by: ensure the sections are blued properly. check blueing reagent. |
| H&E troubleshoot: pH may be above 5 which causes pale cytoplasmic staining. what is it called and how do you prevent it? | Pale Cytoplasmic Staining Prevent by: check eosin solution pH. Completely remove bluing reagent before transferring the slides to eosin. Ensure the sections are not too thin. |
| H&E troubleshoot: cytoplasm may be overstained. what is it called and how do you prevent it? | Dark cytoplasmic staining Prevent by: avoid overconcentrated eosin solution, do not leave eosin too long. Allow sufficient time in dehydrating solutions. Check sections for proper thickens. |
| H&E troubleshooting: Eosin not properly differentiated. | Eosin not properly differentiated. Timely and complete fixation. Good dehydration and clearing during processing. Eosin is at the correct pH. |
| H&E troubleshooting: Blue black precipitate on top sections. what is it called and how do you prevent it? | Blue-black precipitate Prevent by: filtering hematoxylin daily before staining. |
| H&E troubleshooting: Water & slides turn milky when slides are placed in water following alcohol during deparaffinization. what is the cause? and how to prevent it. | Cause: slides & water turn milky after rehydration alcohols, presence of xylene on slides. prevent: back the slides up & change the alcohols. then take slides through fresh absolute & 95% alcohols to water. |
| h&E troubleshooting: slides are hazy or milky in last xylene before applying cover glass. | cause: indicates water is present on slides & dehydration is not complete. Prevent: back slides up & change alcohols & xylenes. then rehydrate the slides in alcohol & clear xylene. |
| h&E troubleshooting: Small patches to large, diffuse areas in sections. | cause: uneven h&e staining prevent by: use toluene instead of xylene if using open processors. check equipment for malfunction. |
| h&E teoubleshooting: Dark basophilic staining of nuclei cytoplasm, especially around tissue edges. | cause: laser and electrocautery techniques denature macromolecules and produce heat artifact. prevent by: there is no remedy to fix artifact. |
| j&E troubleshooting: nucleus& cytoplasm do not contrast well, the cause is from poor staining of the nucleus is too pale, the cytoplasm is overstained, the nuclear stain is too dark or the cytoplasm is too pale. | Cause: Poor contrast between nucleus and cytoplasm prevent by: determine wheater nuclear stain or cytoplasmic stain is inadequate, adjust staining times. Check pH of solutions. monitor pH of water. |
Created by:
Natasha Staley