Troubleshoots

Question 1

Name/ Causes/ remedy

Appear as loss or total disappearance of nuclear chromatin, cell shrinkage, artifactual space around cells, desquamation (loss epithelial cells)

Answer 1

Autolysis (fixation)
Cause: delayed or inadequate fixation
Remedy: placed in fixative ASAP, volume 15-20X, open or slice large specimens, gross thin section, bisect lymph nodes

Question 2

Name/cause/remedy

Appear as separation of tissue components on water bath during microtomy, H&E shows nuclear bubbling or smudgy

Answer 2

Incomplete fixation (fixation)
Cause :
-not enough time in fixative
- fixative depleted causing inadequate fixation prior dehydration

Remedy:
-increase time in fixative
- change to another fixative
- place formal alcohol in first 3 stages of tissue processing
- ensure thin grossed section
- ensure fixative not depleted ( change sol’n frequently)
- don’t pack cassettes tightly

Question 3

Tissues become hard, brittle or shrunken

Answer 3

Dehydration issue from excessive dehydration

Question 4

Prohibit the penetration of the clearing agent into the tissue, leaving the specimen soft and mushy and non-receptive to infiltration

Answer 4

Dehydration issue- incomplete dehydration