Aquitania, Mary Christelle G.

Date Submitted:
UST Medical Technology Intern Histolopathology

Faults occurring during Trimming/Cutting of Paraffin Blocks

Trimming is a process wherein the paraffin block is exposed for actual cutting after when the
wax is solidified and removed from the mold. The sides, top and bottom of the tissue block are
trimmed until leveled perfectly and all sides are parallel to each other. An old knife or blade may
is used for this procedure yet still must be relatively sharp to avoid tissue block damages. After
coarse trimming, the block is then placed in the microtome for final trimming and cutting. To
have a fine thin trimming, setting the thickness adjuster at 15mm or by advancing the block
using coarse feed mechanism is done, according to Gregorios’ Histopathologic Techniques. The
block is clamped on the chuck must be retracted enough to ensure that knife does not touch the
block on initial down stroke. The surface is trimmed first until the tissue surface is partly
exposed. Regular cutting rhythm should be maintained. Thin ribbons are taken out at a time to
prevent the block from cracking. Factor considered for the cutting rate of the tissue blocks are
the type of tissue, size of the slick, and the model of the block. For routine histologic procedures,
usually sections are cut between 4-6μ.

However, during the processing and cutting of the section, several circumstances and difficulties
may be encountered. A medical technologist should be alert taking note of faults, which if not
immediately remedied, shall cause entire failure of process, poor sectioning, and ultimately
improper evaluation of the tissue in question. The most commonly encountered difficulties
observed during tissue processing with its reason and remedy is placed in the table.

FAULTS REASON REMEDY

Prolonged fixation
Prolonged dehydration
Tissue may be softened by
Prolonged clearing
soaking in a small dish
Brittle or hard tissue Prolonged paraffin infiltration
containing water with
Overheated paraffin oven
detergent, phenol or Molliflex
Drying out of tissue before
actual fixation
Water not completely Repeat dehydration with
Clearing agent turns milky as
removed (incomplete absolute alcohol, then repeat
soon as tissue is placed in it
dehydration) clearing
Blocked is trimmed down
nearest to the tissue. The
Clearing agent is not remaining wax id melted on
Upon trimming, tissue smells
completely removed due to embedding oven and paraffin
of clearing agent
insufficient impregnation impregnation is repeated,
changing the paraffin at least
once before blocking.
 Repeat clearing; id object has
already been embedded,
Tissue is opaque, section
prolong oven and paraffin
cutting is difficult due to the Insufficient clearing
impregnations repeated,
presence of alcohol
changing the paraffin at least
once before blocking
Insufficient dehydration,
Tissue shrinks away from wax
therefore incomplete clearing Repeat the whole procedure
when trimmed
and impregnation
Tissue is soft when block is
Incomplete impregnation Repeat whole procedure
trimmed
Air holes on tissue during
Incomplete impregnation Repeat impregnation
trimming
On trimming, wax appears Re-embed in freshly filtrated
Contaminated wax
crystalline wax
Block not cooled rapidly
enough
Paraffin block, after cooling, Insufficient paraffin Repeat paraffin impregnation,
is moist and crumbles impregnation then re-embed

Faults Observed during Section-Cutting

FAULTS REASON REMEDY

Surfaces and edges of the
Re-trim the block
block are not parallel
Horizontal surface of the
Re-adjust and re-orient the
block is not parallel to the
block
knife
Coat horizontal edges of the
Sections fail to form ribbons
Paraffin wax is too hard block with wax of lower
melting point
Knife is tilted to much Reduce the tilt
Readjust the thickness of the
Sections are too tick sections
Hone and strop
Sections roll up on cutting so Knife is blunt Sharpen the knife
that they adhere and get Tilt of knife is too great Reduce the tile
broken against the knife edge Knife edge is dirty Clean the knife edge
Ribbon is curved, crooked or Adjust the knife so that the
Blunt of dull spot on the knife,
uneven instead of straight knife will present a uniformly
producing an irregular knife
sharp edge to the block, or
edge
sharpen
Edges of the block are not Re-trim the block
parallel but round wedge
shape
 Knife is not parallel to the
Readjust knife and block
block
Repeat impregnation using
Paraffin is impure
pure wax
Knife is blunt or dull Re-sharpen the knife
Paraffin block Is warm and Cool the block on ice water
soft until firm
Sections are compressed, Knife edge is coated with
Clean the knife edge
wrinkled or jammed paraffin
Sections are too thin Readjust thickness of section
Microtome set screw is loose Tighten the screw
Tilt of knife is too vertical Reduce the tilt
Sections are squashed (width
Bevel of knife is lost due to Re-sharpen, using a knife back
of each section is less than that
incorrect sharpening or automatic knife sharpener
of block)
Bubble or dirt formed in the Re-embed in freshly filtered
embedding medium wax if necessary
Once embedded in paraffin
A hole is formed in the section
Hard spot in tissue due to wax, decalcification is
calcium impractical; use a base-sledge
microtome with a wedge knife
Tilt of knife is too great or
bevel is not cleared, hence
object is compressed against Reduce the tilt
the knife edge clamp set screw
on knife
Section of unequal thickness
Or blockholder is loose
are produced Tighten the screw
Blocks are too large
Cut blocks into smaller
Block are too hard
fragments
Soften the blocks in detergent
or phenol
Breather out or blow gently on
the block and knife to breakup
Static electricity due to low
static electricity, or boil water
atmospheric humidity
Sections adhere to the knife or in the room to increase the
other parts of the microtome humidity
Knife edge is dirty Clean the knife edge
Knife edge is dull Sharpen the knife
Knife tilt is too great Reduce the tilt
Nicks or damage on the knife
Sharpen the knife
Ribbon is split or lengthwise edge
vertical scratches are seen on Dirty embedding Re-embed in filtered wax
sections Knife edge is dirty Clean knife edge with xylene
Tilt of knife is too great Reduce the tilt
Sections are lifted from the Knife tilt is too great Reduce the tilt
 Knife is dull Sharpen the knife
knife on upstrokes Paraffin is too soft or room
Cool paraffin wax in ice water
temperature is warm
Tilt of knife is too small,
Resistance is felt on the lower paraffin block is therefore
part of the section during compressed against the base of Increase the tilt
cutting the knife towards the end of
stroke
Horizontal or parallel lines or Knife edge vibrate due to Treat with phenol during
furrows across the section hardness of tissue processing or collodionize
(“Chatters”) are seen, forming
Tilt of knife is too great Reduce the tilt
thin and thick zones
Knife is blunt Sharpen knife
Knife is not clamped properly Adjust the knife
Tilt of knife is too great Reduce the tilt
Section cut is sometimes thin, Tighten adjusting and locking
Knife or block holder is loose
sometimes thick screws
Knife tilt is too small that
block is compressed by bevel Increase the tilt
and section is not cut
Tilt of knife is too slanted or Readjust the angulation of the
Knife makes a hard metallic
too big knife
scrapping or ringing sound on
Take fresh block treated with
backstroke, when section is Tissue is too hard
phenol during processing
cut
Knife blade is too thin Change the knife
Frozen tissue crumbles and
comes off the block holder Freezing is not adequate Refreeze the tissue block
when cut
Frozen tissue chips into
Tissue is frozen too hard Warm the tissue with fingers
fragments when cut