HISTOPATHOLOGY

RECEIVING/RELEASING
MOUNTING
RECEIVING
• Logbook
1. Biopsy
2. Pap Smear
3. Peripheral Blood Smear (Adult, Pedia)
4. Cell Block
5. FNAB
- Name
- Age/Gender
- Ward
- Specimen
- Billing
RECEIVING
• Claim Stub
- Name - Date Received
- Date Due - Lab Number
Designations:
• SP – Surgical Pathology
• CYP – Cytocolor Procedure
• HP – Histopathologic Procedure
• CB – Cell Block
• GA – General Autopsy
• DP – Dermatologic Procedure
• FNAB – Fine Needle Aspiration Biopsy

• Encoding
RELEASING
• Logbook
• Find the lab number of the patient, if there is already a check mark,
the result has already been endorsed either to the pathologists’
office, on the result folders, or on the computer table.
• If there is no check, the result is still pending. It’s either there is still
no pocket of slides or the pocket of slides is in the pathologists’
office ready for reading. To be sure, just ask the staff on the
measures to be taken.
• If you’ve already seen the result form and there is no signature of
the pathologist, put a stamp of the corresponding name of the
pathologist, find the pocket of slides on the cabinet below the pile
of logbooks, and bring them to the pathologists’ office for
verification and signature.
RELEASING
NOTE!!!
- DO NOT ENTER IF THERE ARE CONSULTANTS PRESENT.
- Any pathologist can sign the result form as long as there
is already a stamp.
- If there are corrections, endorse the result for re-typing.
Automated processing
Reagent preparation
AUTOMATED PROCESSING
• This method makes use of an automatic tissue
processing machine which fixes, dehydrates, clears and
filtrates tissues thereby decreasing the time and labor
needed during the processing of tissues, resulting in a
more rapid diagnosis with less technicality.
• Constant agitation (vertical and rotation) which
accelerates and improves tissue penetration giving rise
to more consistent results.
• Elliott-Bench Type Processor
• PROCEDURES
• Dehydration
• 95% ethyl alcohol – 30 minutes
• 95% ethyl alcohol – 30 minutes
• Acetone 1 – 30 minutes
• Acetone 2 – 30 minutes
• Acetone 3 – 30 minutes
• Clearing
• Xylene 1 – 1 hour
• Xylene 2 – 1 hour
• Infiltration
• Paraffin 1 – 1 hour
• Paraffin 2 – 1 hour
Reagent Preparation
• Acid-alcohol and Ammonia water c/o staff
• Xylene, Acetone and paraffin wax readily available (if not, make a
request)
• 95% Ethyl Alcohol
• 950ml absolute ethanol + 50ml distilled water
• If 80% (800ml+200ml)
• If 70% (700ml+300ml)
• 10% Neutral Buffer Formalin
• 39g of disodium hydrogen phosphate + 24g sodium dihydrogen phosphate
dehydrate + <630ml 37% formalin
• Addition of tap water; c/o Staff
• Checking of pH, a day after the formalin was made (inventory)
EMBEDDING and TRIMMING
EMBEDDING
• The tissues are positioned in a mold or block of
the infiltrating medium and are prepared for
section cutting
TYPES OF BLOCKING MOLD
• LEUCKHART’S EMBEDDING MOLD

• COMPOUND EMBEDDING UNIT

• PLASTIC EMBEDDING RINGS AND BASE MOLD

• DISPOSABLE EMBEDDING MOLDS

PLASTIC EMBEDDING RINGS AND
BASE MOLD
 ORIENTATION

• Process by which a tissue is arranged in precisely arranged

positions in the mold during embedding, on the microtome
before cutting and in the slide before staining
POINTS TO CONSIDER:
• Elongated tissues are placed diagonally across the
block

• Tubular specimen and gastrointestinal tissues are

embedded en face so as to provide cross sections
showing all tissue layers
• Tissues with an epithelial surface such as
skin are embedded to provide sections in a
plane at a right angle to the surface
• Multiple tissue pieces are aligned
across the long axis and the center
of the mold, and not placed
randomly
 TRIMMING
• Process of removing excess wax after embedding. The wax is
cut off to from the block expose the tissue surface in
preparation for actual cutting
 Procedure
Distilled/Tap water 10 dips
Hematoxylin 1 minute
Wash in running water 1 minute
2-propanol 10 dips
Polychromic stain 1-5 minutes
Wash in water 10 dips
2- propanol 10 dips
2-propanol 10 dips
 -used to demonstrate
Mycobacterium leprae (leprosy)
 PROCEDURE
Dip in xylene 15 minutes
Dip in peanut oil 24 minutes
Blot dry in filter paper
Dip in xylene 5 minutes
Air dry.
Flood with carbolfuchsin 30 minutes
Wash with tap water 10 dips
Dip in acid alcohol Until faint pink
Wash in tap water
Flood with methylene blue 10 minutes
Wash in tap water
Air dry.
MOUNTING

- AFTER STAINING AND BEFORE EXAMINATION,

SECTIONS SHOULD BE MOUNTED W/ A SUITABLE MEDIUM
UNDER COVER SLIPS.

PURPOSE:
- TO FACILITATE EASE OF HANDLING AND STORAGE AND
PREVENT DAMAGE TO THE SECTION.
• MATERIALS NEEDED: DROPPER/PASTEUR PIPPETTE, MOUNTING
MEDIA, COVERSLIP(LARGE & SMALL), TRAY W/ PAPERS

• SPX SLIDES TO BE MOUNTED: FNAB, CELL BLOCK, SP, FITE FARACO,

AND OTHER BIOPSY SPX.

• SPX SLIDES NOT TO BE MOUNTED: CYP(PAP SMEAR)

• REFER TO THE INTSRUCTIONS POSTED AT THE MOUNTING AREA
PROCEDURE:
• LET THE STAFF CHECK THE AIR-DRIED SLIDES, BEFORE CLEANING THE
SLIDES.
• CLEAN THE SLIDES W/ A GAUZE PAD CONTAINING ETHANOL.
• PLACE 2-3 DROPS OF MOUNTANT ON THE SURFACE OF THE SECTION.
• PLACE THE COVERSLIP OVER THE MOUNTANT.
• LET THE MOUNTANT SPREAD SMOOTHLY BETWEEN THE COVERLIP
AND SLIDE.
• WHAT TO AVOID:
• Presence of bubbles
• Excess mountant
• Contaminant
• Dirty way of mounting

Remedies:
- Bubbles can be removed by aspirating it with the pipet or dropper
- A dirty mounted slide can be cleansed w/ xylene (dip in xylene for
some time)