What;s Wrong?
Quiz yourself by thinking what should be in
each of the black spaces below before clicking
on it to display the answer.
Help!
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| 1. Problem: White spots are seen in the section after deparaffinzation. (#1) | Cause #1: The slide was not dried properly before beginning deparaffinization. Remedy #1: Put slide in absolut alcohol to remove water then into xylene.
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| 1. Problem: White spots are seen in the section after deparaffinzation. (#2) | Cause#2: The slide did not remain in xylene long enough to complete the removal of paraffin. Remedy #2: Put slide back into xylene for a longer period of time.
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| 2. Problem: Pale nuclei (hematoxylin is too light) *Causes | Causes: Not stained long enough, stain overoxidized (stain too old), differentiated too long, bones overdecalcified.
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| 2. Problem: Pale nuclei (hematoxylin is too light) *Remedies | Remedy: The section must be restained, If fixed in Zenker's, increase staining time or use a method to increase basophilia.
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| 3. Problem: Nuclei are overstained (hematoxylin too dark) | Cause #1: Stained too long. Remedy #1: Decolorize the section and restain. Cause #2: Sections are too thick. Remedy #2: Resection the tissue.
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| 4. Red or red-brown nuclei. | Cause #1: Hematoxylin has broken down. Remedy #1: Check the oxidation status of the hematoxylin. Cause #2: Slides not blued sufficiently. Remedy#2: Allow for a longer bluing time. You cannot overblue sections.
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| 5. Problem: Pale staining with eosin. *Causes | Causes: pH of eosin is over 5.0 (ammonia water), sections too thin, dehydrated too long.
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| 5. Problem: Pale staining with eosin. *Remedies | Remedies: Check the pH and readjust it with acetic acid to 4.5-5.0, resection tissues, make sure that slides do not spend too much time in the lower concentration alcohols as water will decolorize the eosin.
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| 6. Problem: Cytoplasm is overstained. *Causes | Causes: Sectioned too long, Eosin too concentrated (dilute), sections too thick, dehydrated too quickly.
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| 6. Problem: Cytoplasm is overstained. *Remedies | Remedies: Dilute eosin or decrease staining time, resection the tissue, if not well differentiated, rehydrate slides and then dehydrate them again.
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| 7. Blue-Black precipitate on top of slide | Cuase: Metal sheen on the top of the unfiltered hematoxylin pisked up on the slide. Remedy: Filter the hematoxylin (daily)
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| 8. Problem: Water bubbles are seen microscopically in the stained sections. | Cause: Sections were not completely dehydrated. Remedy: Remove cover glass and mounting media with xylene. Place the slide in fresh absolute alcohol then back into xylene and coverslip.
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| 9. Problem: Difficulty bringing some areas of the tissue in focus with the light microscope. | Cause: Mounting media may be on top of the cover glass. Remedy: Remove the cover glass and remount with a new cover glass.
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| 10. Problem: Mounting media has retracted from the edge of the coverslip. | Cause: The cover glass is warped or mounting media has been thinned by too much xylene. Remedy: Remove cover glass and apply new cover glass with fresh mounting media.
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| 11. problem: The water and the slides turn milky when the slides are placed in the water following the rehydrating alcohols. | Cause: Xylene has not been removed completely by the alcohols. Remedy: Change the alcohols and back up to absolute alcohol and dehydrate the sections.
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| 12. Problem: The slides are haxy or milky in the last xylene rinse prior to coverslipping. | Cause: Water has not been completely removed from the sections before being placed into zylene. Remedy: Change the alcohol solutions, especially the absolute alcohol. Redehydrate the sections and clear in fresh xylene.
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| 13. Problem: |
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| 9. Problem: Difficulty bringing some areas of the tissue in focus with the light microscope. | Cause: Mounting media may be on top of the cover glass. Remedy: Remove the cover glass and remount with a new cover glass.
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| 10. Problem: Mounting media has retracted from the edge of the coverslip. | Cause: The cover glass is warped or mounting media has been thinned by too much xylene. Remedy: Remove cover glass and apply new cover glass with fresh mounting media.
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| 11. problem: The water and the slides turn milky when the slides are placed in the water following the rehydrating alcohols. | Cause: Xylene has not been removed completely by the alcohols. Remedy: Change the alcohols and back up to absolute alcohol and dehydrate the sections.
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| 12. Problem: The slides are haxy or milky in the last xylene rinse prior to coverslipping. | Cause: Water has not been completely removed from the sections before being placed into zylene. Remedy: Change the alcohol solutions, especially the absolute alcohol. Redehydrate the sections and clear in fresh xylene.
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| 13. Problem: |
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