Answer The Following:: Module:1 Laboratory Techniques in Pathology Lab

NGUYEN, DUC Q 26 MAY 2022
DENT 3k | GENERAL PATHOLOGY LABORATORY
Module:1 Laboratory Techniques in Pathology Lab
Answer the following:

A. How are paraffin sections prepared in the lab? Provide a step by step procedure using images with brief descriptions.
A. OBTAINING A FRESH SPECIMEN
Fresh tissue samples will be obtained from a variety of sources. It should be noted that they can very easily be damaged during
removal from the patient or experimental animal. It's important that they're handled with care and mended properly as soon as
possible after dissection.
B. FIXATION
A liquid fixative, such as formaldehyde solution, is where the specimen is placed. This will slowly penetrate the tissue, creating
chemical and physical changes that will harden and preserve the tissue, as well as protect it from further processing steps. There
are only a few chemicals that can be used for fixation since they must have specific qualities.
C. DEHYDRATION
Because melted paraffin wax is hydrophobic (insoluble in water), the majority of the water in a specimen must be eliminated
before wax can be infiltrated. Immersing specimens in a series of ethanol (alcohol) solutions of increasing concentration until
pure, water-free alcohol is obtained. Ethanol is miscible with water in all quantities, therefore the alcohol gradually replaces the
water in the specimen. To minimize significant tissue distortion, a series of increasing concentrations is used.
D. CLEARING
Despite the fact that the tissue is now essentially water-free, we are still unable to infiltrate it with wax due to the fact that wax
and ethanol are highly incompatible. As a result, we'll need an intermediate solvent that's completely miscible with both ethanol
and paraffin wax. The ethanol in the tissue will be displaced by this solvent, which will then be displaced by molten paraffin
wax. The cleansing agent also removes a significant amount of fat from the tissue, which would otherwise act as a barrier to
wax entry. Xylene is a common clearing agent, and it takes several changes to completely replace ethanol.
E. WAX INFILTRATION
A suitable histological wax can now be infiltrated into the tissue. At 60°C, a common wax is liquid and can be infiltrated into
tissue, then cooled to 20°C, where it solidifies to a consistency that permits pieces to be cut consistently. These waxes are made
out of purified paraffin wax and a variety of additives, such as styrene or polyethylene resins. These wax formulations have
very specific physical properties that allow tissues infiltrated with the wax to be sectioned at a thickness of at least 2µm, to
form ribbons as the sections are cut on the microtome, and to retain enough elasticity to flatten completely during flotation in a
warm water bath.
F. EMBEDDING OR BLOCKING OUT
The specimen must now be formed into a "block" that can be clamped into a microtome for section cutting after it has been
thoroughly infiltrated with wax. An "embedding centre" is used for this phase, where a mold is filled with molten wax and the
specimen is placed within. The specimen is properly oriented in the mold because the "plane of section," an important
consideration in both diagnostic and research histology, is determined by its positioning. A cassette is placed on top of the
mold, which is then filled with more wax and set on a cold plate to harden. The block, together with its attached cassette, can
now be withdrawn from the mold and ready for microtomy. It's important to note that, if tissue processing is done correctly, the
wax blocks containing the tissue specimens are very stable and can represent an important source f archival material
B. Download an image of a tissue sample with the following stains. Provide information on the images
A. H&E
• Skin H&E. Note the balanced coloration in this section of skin. The nuclei are stained purple, while the cytoplasmic
components are pink.
B. CONGO RED
• Congo red stained salivary gland section examined under crossed polarized light from patient with AL amyloid imaged
using clinical microscope and same field examined using Metallurgical microscope
C. ZIEHL NEELSEN
• Ziehl-Neelsen stain of the purulent soft tissue specimen revealing pleomorphic bacilli.
REFERENCES:
https://www.leicabiosystems.com/knowledge-pathway/an-introduction-to-specimen-processing/
https://paraffinwaxco.com/paraffin-tissue-block/
https://www.leicabiosystems.com/knowledge-pathway/he-staining-overview-a-guide-to-best-practices/
https://diagnosticpathology.biomedcentral.com/articles/10.1186/s13000-019-0822-4
https://www.researchgate.net/figure/Ziehl-Neelsen-stain-of-the-purulent-soft-tissue-specimen-revealing-pleomorphic-
bacilli_fig2_323954567

Answer The Following:: Module:1 Laboratory Techniques in Pathology Lab

Uploaded by

Copyright:

Available Formats

Answer The Following:: Module:1 Laboratory Techniques in Pathology Lab

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Answer The Following:: Module:1 Laboratory Techniques in Pathology Lab

Uploaded by

Copyright:

Available Formats

NGUYEN, DUC Q 26 MAY 2022

DENT 3k | GENERAL PATHOLOGY LABORATORY

Module:1 Laboratory Techniques in Pathology Lab

Answer the following:

F. EMBEDDING OR BLOCKING OUT

You might also like