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Activity For Day 2

This document provides guidance on troubleshooting various histopathology techniques. It discusses possible reasons for thick and thin tissue sections, crooked ribbons, sections that won't ribbon, and solutions for over-fixed or decalcified tissue. For tissue that is sludgy, mushy or greasy, the likely cause is lipids or fats and the solution is to remove excess wax and rehydrate the tissue in hot saline before reprocessing. "Sweating" during flotation can be caused by insufficient dehydration and clearing, and decalcifying hard fragments before re-sectioning can produce a better slide. Rehydration solutions like Sandison's solution can be used to treat over-processed or dried

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Catherine Merilleno
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77 views4 pages

Activity For Day 2

This document provides guidance on troubleshooting various histopathology techniques. It discusses possible reasons for thick and thin tissue sections, crooked ribbons, sections that won't ribbon, and solutions for over-fixed or decalcified tissue. For tissue that is sludgy, mushy or greasy, the likely cause is lipids or fats and the solution is to remove excess wax and rehydrate the tissue in hot saline before reprocessing. "Sweating" during flotation can be caused by insufficient dehydration and clearing, and decalcifying hard fragments before re-sectioning can produce a better slide. Rehydration solutions like Sandison's solution can be used to treat over-processed or dried

Uploaded by

Catherine Merilleno
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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Jericho M.

Merilleno
4DMT

ACTIVITY FOR DAY 2

1. Below are problems in histopathologic technique. For each situation, give possible
reasons that might cause the problem.
a. Cutting thick and thin sections
This can happen to alternate section, or it can happen within the same
section.

This is a section of uterus stained Hematoxylin and Eosin and showing thick and
thin areas. The thick area, of course, stains more darkly than the thin area.
Why does this happen?
i. It may be due to a loose block
ii. Insufficient clearance angle
iii. Less than ideal paraffin wax
b. Crooked or uneven ribbons when sectioning. What causes this to happen?
i. Non parallel block edges
ii. Dull blade
c. Sections will not ribbon. Why?
i. Debris on knife
ii. Incorrect clearance angle
iii.
2. Below are problems in histopathologic technique. For reach situation, give a possible
solution?
a. How to reestablish nuclear staining in tissue sections that have been over-
fixed or decalcified?
Use a solution that allows fixation and decalcification to occur
simultaneously.
b. Tissue Excessively Dehydrated
Decrease dehydration time of the tissue specimen

3. The St. Rock Cancer Institute has been the topmost research center in the area.
They receive multiple organs per day to be processed and analyzed. You were hired
as the new clinical laboratory scientist assigned to process breast samples at that
month. Breast samples are processed both thru routine histopath and
immunohistochemistry staining. On your first day of duty, you notice that breast
tissues are sludgy, mushy or greasy and the tissue slides produced has a rim only of
fragmented tissue. To test your skill, the section head requested you to investigate
on it. What could be the possible explanation to it? How could this situation be

resolved?
Sludgy, mushy, or greasy tissues are caused by lipids or fats from the tissue sample.
This can be resolved by removing excess wax and the tissue be rehydrated using hot
saline before reprocessing
4. Image shown below is an example of “Sweating” during flotation. It is described as a
translucent appearance sometimes seen in brain and spinal cord blocks. Explain
possible causes of this scenario and ways to prevent it.

This might be due to insufficient dehydration and clearing which results to


sweating which may cause oil droplets

5. In the histopathology section of St. James Hospital, a tissue with hard fragments
was processed. The tissue slide that contains granuloma (H & E) was processed
and shows a microscopic image as shown below. Much of it has fragmented and
fallen from the section while the remainder was badly shredded. What could be the
possible ways to troubleshoot and produce a better tissue slide? Discuss the reason
behind the basophilic area pointed in the image.

The agent responsible is most likely to be calcium. This can be resolved by decalcifying
the surface of the sample to be followed by washing and re-sectioning of the block.
6. Tissue samples described as cooked tissue is usually over-processed (or has been
subjected to extreme conditions e.g., excessive heat). The tissue section of tongue
before and after reconstitution is shown in the image below. Discuss the use of
rehydration solutions like Sandison’s solution and steps on how to use it to
troubleshoot this tissue sample.

Rehydration solutions are used on tissue samples that are dried or exposed too
much in air for a long period of time. The tissue sample should be submerged in
Sadinson’s solution for 12 to 18 hours to rehydrate the tissue.

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