Question 1

1. Problem: White spots are seen in the section after deparaffinzation. (#1)

Accepted Answer

Cause #1: The slide was not dried properly before beginning deparaffinization. Remedy #1: Put slide in absolut alcohol to remove water then into xylene.

Question 2

1. Problem: White spots are seen in the section after deparaffinzation. (#2)

Accepted Answer

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.

Question 3

2. Problem: Pale nuclei (hematoxylin is too light) *Causes

Accepted Answer

Causes: Not stained long enough, stain overoxidized (stain too old), differentiated too long, bones overdecalcified.

Question 4

2. Problem: Pale nuclei (hematoxylin is too light) *Remedies

Accepted Answer

Remedy: The section must be restained, If fixed in Zenker's, increase staining time or use a method to increase basophilia.

Question 5

3. Problem: Nuclei are overstained (hematoxylin too dark)

Accepted Answer

Cause #1: Stained too long. Remedy #1: Decolorize the section and restain. Cause #2: Sections are too thick. Remedy #2: Resection the tissue.

Question 6

4. Red or red-brown nuclei.

Accepted Answer

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.

Question 7

5. Problem: Pale staining with eosin. *Causes

Accepted Answer

Causes: pH of eosin is over 5.0 (ammonia water), sections too thin, dehydrated too long.

Question 8

5. Problem: Pale staining with eosin. *Remedies

Accepted Answer

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.

Question 9

6. Problem: Cytoplasm is overstained. *Causes

Accepted Answer

Causes: Sectioned too long, Eosin too concentrated (dilute), sections too thick, dehydrated too quickly.

Question 10

6. Problem: Cytoplasm is overstained. *Remedies

Accepted Answer

Remedies: Dilute eosin or decrease staining time, resection the tissue, if not well differentiated, rehydrate slides and then dehydrate them again.

Question 11

7. Blue-Black precipitate on top of slide

Accepted Answer

Cuase: Metal sheen on the top of the unfiltered hematoxylin pisked up on the slide. Remedy: Filter the hematoxylin (daily)

Question 12

8. Problem: Water bubbles are seen microscopically in the stained sections.

Accepted Answer

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.

Question 13

9. Problem: Difficulty bringing some areas of the tissue in focus with the light microscope.

Accepted Answer

Cause: Mounting media may be on top of the cover glass. Remedy: Remove the cover glass and remount with a new cover glass.

Question 14

10. Problem: Mounting media has retracted from the edge of the coverslip.

Accepted Answer

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.

Question 15

11. problem: The water and the slides turn milky when the slides are placed in the water following the rehydrating alcohols.

Accepted Answer

Cause: Xylene has not been removed completely by the alcohols. Remedy: Change the alcohols and back up to absolute alcohol and dehydrate the sections.

Question 16

12. Problem: The slides are haxy or milky in the last xylene rinse prior to coverslipping.

Accepted Answer

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.

Question 17