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Histopathology in Practice
Sample Reception, Dissection,
Processing, Embedding.
Della Thomas
Senior Biomedical Scientist
Independent Histopathology Services
della.thomas@unilabs.com
Histology
The science concerned with the microscopic
structure of tissues and organs in relation to
their function.
Also called microanatomy.
Specimen Reception
Specimen Unpacking
Specimen QC

Mickey Mouse
Rt nasal polyp
DOB:

MOUSE
Mr. MICKEY

18/11/28
26/11/13

Dr. Makewell

Rt nasal polyp
C/O loss of
function - ↓
ability to sniff
out cheese
O/E: nasal polyp
? nature.

18/11/28
123456

Dr. Makewell
26/11/13 8am

M
Specimen Labelling
Sample documenting
Assign the
specimen to a
pathologist

MOUSE
Mr. MICKEY
Rt nasal polyp
C/O loss of
function - ↓
ability to sniff
out cheese
O/E: nasal polyp
? nature.

18/11/28
123456

Dr. Makewell
26/11/13 8am

M
Cassetting
Specimen Dissection - the stage
Dissecting props
•
•
•
•
•
•
•
•
•

Cutting board
Forceps
Ruler
Scalpel / knives / saw
Inks / dyes
Cassettes / lids
Biopsy pads / tissue
Filter bags
Weighing scales
Cut-up dictation
• “Morphology” computer
system
• “Dragon” dictation system
• Hands-free
• Just scan the barcode and
away we go!!
Specimen Categories

A
B
C

Specimens only requiring transfer from container
to tissue cassette.

D
E

Dissection and sampling required needing a
moderate level of assessment.

Specimens requiring transfer but with standard
sampling, counting, weighing or slicing.
Simple dissection required with sampling
needing a low level of diagnostic assessment
and/or preparation.

Specimens requiring complex dissection and
sampling methods
Category A Specimens
Breast core
biopsies

Endometrium

Colonic series
Category B Specimens

Small lipoma

Small skin
biopsy
Cervical LLETZ
Category C Specimens
Prepuce

Gallbladder

Haemorrhoids

Appendix
Category D Specimens
Pigmented skin
lesions

Large Intestine
(Crohn’s)

C/D
C/D

Skin with markers

Salivary gland
tumour
Category E Specimens
Breast cancer

Thyroid (medullary
Ca)
Testis (seminoma)

Uterus (endomet. Ca)
Specimen Dissection
What does the pathologist
need to know?
Provide good descriptions - say what you see!!
• Shape
• Colour
• Texture
• Dimensions
• Weight
• Distances from margin(s)
• Orientation markers
• Cut surface appearance
etc....
• Keep your fingers crossed for good clinical history
Doctors’ handwriting!!
“dysplasia”?
OR “lymphoma”?

Breast Ca....”Steroids”?
OR “Stomach”?

“Nothing” suspicious
OR “Notably” suspicious?
Simple transfer

Cervical biopsy

Prostate cores
Specimen Sampling
• Literally, taking a sample of the tissue
• Representative
• Generally, fewer blocks required if the tissue
looks uniform throughout (for benign cases)
• Sampling “rules”
Sampling rules: prostate chips
• If the chippings weigh 12g
or less, the entire
specimen must be
processed
• If the chippings weigh
more than 12g, a minimum
of 6 cassettes must be
processed

Prostate chips (19g)

Sampled in 8 cassettes:
First 12g = 6 cassettes

• For every 5g over 12g, one
more cassette must be
processed

19g = 7g over 12g
1 cassette per 5g over = 2
more cassettes.
Simple dissection
Specimen:
Skin from back
Clinical details:
Sebaceous cyst
Simple dissection
Unlock the pathology
Fistula-in-ano
Unlock the pathology

Gallbladder
Unlock the pathology
Cystoprostatectomy

Posterior view
Inking
• Resection margins
• Embedding instructions
• Orientation
• Distinguish between samples
• Identify the cut surface
• Acetic Acid
Inking resection margins
Pigmented lesion

Cervical cone
Embedding Instructions

Ink dots instruct the embedder to embed
the tissue a certain way.
Orientation - which way up?
• Anatomical
Orientation - which way up?
• Clockface
3 o’clock marker
Clinical Data:
Suture marker
at 3 o’clock.
“12 o’clock” marker
Clinical Data:
Tag suture at
12 o’clock.

?
“12 o’clock” marker
Mapping on a larger scale
Cystoprostatectomy
Prostate

Ureter

Red = Right
Green = Left

Posterior Anterior
bladder bladder
Cassette sizes
Calcified / firm tissue
Most common:

• Femoral head
• Bone Marrow Trephine
• Ethmoid mucosa / nasal polyps (cartilage)
• Nail
• Hardened cysts
Softening
• For bony / hard tissue:
10% Formic Acid
• For nail:
Phenol or hair removal
cream.
Firm tissue testing methods
• X-ray - Expensive / ? bench space, but
very accurate
• Chemical end-point test (Ammonium
Hydroxide/Ammonium Oxalate) - very
time consuming, but accurate
• Physical manipulation - not very
accurate, may damage the specimen,
but simple and inexpensive
Cutting instructions
•
•
•
•
•
•
•

Levels
Special stains
Unstained Sections
Serial sections
Alopecia protocol
MM sentinel lymph node
Hirschprung’s protocol
Specimen Storage
• Ventilated storage units
• Largest buckets lower shelves
• Units are in date (week) order
• 5-weeks’ worth of storage
• Only authorised specimens
are discarded after 5 weeks
• Any outstanding cases are
stored separately until further
notice
Processing
Formalin → Alcohol → Xylene → Wax
Wax

Xylene

Formalin

Alcohol
Processing programs
Embedding
The immortalisation of tissue presentation
Embedding Centre
Molten wax
Used lids

Heated chambers

Wax flow adjuster
Hot surface

Cold plates

Wax dispenser
Embedding tools
Embedding process
Dispense wax

Cool in place

Align tissue

Cassette on
Embedding process
ID bead

Top-up wax

Leave to set

Cool plate
Embedding tips
• Always keep your eye on the tissue:
even for the most careful embedders,
tissue can ping like tiddlywinks. Make
sure you see where it lands!!
• Ink dots: usually used to instruct the
embedder to embed the tissue a specific
way. Make sure you know your own
lab’s protocol.
• Hide-and-seek: open lids / sponges
carefully - tissue often sticks to them.
~ why it’s important for the embedder to
know number of bits in the cassette
• Cleanliness: always watch out for potential carry-over!!
Final block preparation
Imagination
Thank you
www.slideshare.net/DellaThomas
della.thomas@unilabs.com
www.micro2tele.com
@dellybean (Della Thomas)
@HistoQuarterly (Histology Blog)
www.facebook.com/histoquarterly

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