Which step comes first in staining frozen sections with H&E?
The majority of cell and tissue components have no natural color and are not visible. The first step in performing an H&E stain is to dissolve all the wax away with xylene (a hydrocarbon solvent).
How do you stain a frozen section?
Sample Toluidine blue staining method:
- Alcoholic formalin (10 dips)
- Tap water (10 dips)
- Tap water (10 dips)
- 1% T-blue (40 seconds)
- Tap water (10 dips)
- 70% ethanol (10 dips)
- 95% ethanol (10 dips)
- 100% ethanol (10 dips)
How a frozen sample can be processed for staining?
When ready to stain, remove slides from freezer and warm to -20°C in the cryostat or -20°C freezer, fix for 2 minutes in cold fixative (acetone or other suitable fixative) and allow to come to RT to continue with the staining.
How do you Destain H&E slides?
Clinically, we de-stain H&E slides as a regular protocol using 1:1 Xylene/Acetone solution to remove the coverslip and 1% Acid (HCl) Alcohol to remove the stains. Acid alcohol is used in regressive hematoxylin staining regularly and eosin is quickly removed by both acetone and alcohol.
Can you do H&E on frozen sections?
Hematoxylin and eosin stain on frozen sections for Laser Capture microscopy. immerse in eosin for 90 seconds. dehydrate sections with two 10 second washes in 95% ethanol and two 10 second washes with 100% ethanol. place in xylene for 30 seconds.
How long does a frozen section take?
A frozen section usually takes between 30 and 45 minutes. Sometimes the frozen section approach is non-diagnostic or inconclusive because only a limited number of sections can be taken and examined by the pathologist in the short time available.
What is the importance of frozen sections?
A frozen section is a term referring to a section of tissue that has been rapidly cooled using cryostat. It is an important feature that is needed in hospitals to assist with the diagnoses of lesions and the extent of the lesion during surgery.
What are some of the common problems encountered when doing the staining techniques?
Some Common Problems Encountered in H&E Staining
- Nuclear Staining Errors.
- Poor Eosin Differentiation.
- Poor Contrast Between the Hematoxylin and the Eosin.
- Reddish- Brown Nuclei.
- Dark Precipitate On The Slide.
- Uneven Staining.
- White Patches on Slides After Deparaffinization Step.
- Clear Patches on Tissue After Hydrating.
How do you prepare frozen tissue sections?
Prepare frozen tissue sections (steps 1-8): Slowly place the base mold containing the tissue block into liquid nitrogen till the entire tissue block is submerged into liquid nitrogen to ensure tissue is frozen completely. Store the frozen tissue block at -80°C until ready for sectioning.