FAULTS OBSERVED IN TISSUE PROCESSING

FAULTS
REASON
REMEDY
Brittle or hard tissue Prolonged fixation
Tissue may be softened by
Prolonged dehydration
soaking in a small dish or
Prolonged clearing
bowl containing water with
Prolonged paraffin
detergent, phenol, or
infiltration
molliflex
Overheated paraffin oven
Drying out of tissue before
actual fixation
Clearing agent turns Water not completely
Repeat dehydration with
milks as soon as
removed due to incomplete absolute alcohol, then
tissue is placed in it
dehydration
clear again
On trimming, tissue
Clearing agent not
Block is trimmed down
smells of clearing
completely removed due to nearest to the tissue.
agent
insufficient impregnation
Remaining wax is melted
on embedding oven and
paraffin impregnation is
repeated, changing the
paraffin at least once
before blocking
Tissue is opaque,
Insufficient clearing
Repeat clearing; if object
section cutting is
has already been
difficult due to
embedded, prolong
presence of alcohol
clearing up to 12 hours the
re-embed
Tissue shrinks away
Insufficient dehydration
Repeat the whole
from wax when
therefore incomplete
procedure
trimmed
clearing and impregnation
Tissue is soft when
Incomplete fixation
Repeat the whole
block is trimmed
procedure
Air holes found on
tissue during
trimming
On trimming, wax
appears crystalline
Paraffin block, after
cooling, is moist and
crumbles

Incomplete impregnation

Repeat impregnation

Contaminated wax
Block not cooled rapidly
enough
Insufficient paraffin
impregnation

Re-embed in freshly
filtered wax

FAULTS OBSERVED IN TISSUE CUTTING

FAULTS
REASON
REMEDY
Section fail
Surfaces and Re-trim the
to form
edges of the
block
ribbons
block are not
parallel
Horizontal
Readjust and
surface of the re-orient the
block is not
block
parallel to the
knife
Paraffin wax
Coat
is too hard
horizontal
edges of the
block with wax
of lower
melting point
Knife is tilted Reduce the tilt

Repeat paraffin
impregnation, then reembed

too much
Sections are
too thick
Knife is dull
Section roll
up on
cutting so
that they
adhere and
get broken
against the
knife edge
Ribbon is
curved,
crooked, or
uneven
instead of
straight

Knife is blunt
Tilt of knife is
too great
Knife edge is
dirty

Blunt of dull
spot on the
knife,
producing an
irregular
knife edge

Edges of
block are not
parallel but
round or
wedge shape
Knife is not
parallel to the
block
Paraffin is
impure

Sections
are
compressed
, wrinkled,
or jammed

Sections
are
squashed
(width of
each
section is
less than
that of
block)
Hole is
formed in
the section

FAULTS

Knife is blunt
or dull
Paraffin block
is warm and
soft
Knife edge is
coated with
paraffin
Sections are
too thin
Microtome
set screw is
loose
Tilt of knife is
too vertical
Bevel of knife
is lost due to
incorrect
sharpening

Readjust the
thickness of
the sections
Hone and
strop
Sharpen the
knife
Reduce the tilt
Clean the
knife edge

Adjust the
knife so that
knife edge will
present a
uniformly
shard edge to
the block, or
sharpen
Re-trim the
block

Readjust the
knife and
block
Repeat
impregnation
using pure
wax
Re-sharpen
the knife
Cool the block
on ice water
until firm
Clean the
knife edge
Readjust
thickness of
section
Tighten the
screw
Reduce the tilt
Re-sharpen
using a knife
back or
automatic
knife
sharpener

Bubble or dirt
formed in the
embedding
medium

Re-embed in
freshly filtered
wax if
necessary

REASON
Hard spot in

REMEDY
Once

tissue due to
calcium

Sections of
unequal
thickness
are
produced

Tilt of knife is
too great or
bevel is not
cleared,
hence object
is
compressed
against the
knife edge
Clamp set
screw in knife
or block
holder is
loose
Blocks are
too large
Blocks are
too hard

Sections
adhere to
the knife or
other parts
of the
microtome

Ribbon is
split or
lengthwise
vertical
scratches
are seen on
sections

Sections
are lifted
from the
knife on
upstrokes

Resistance
is felt on

Static
electricity
due to low
atmospheric
humidity

Knife edge is
dirty
Knife edge is
dull
Knife tilt is
too great
Nicks or
damage on
the knife
edge
Dirty
embedding
Knife edge is
dirty
Tilt of knife is
too great
Knife tilt is
too great
Knife is dull
Paraffin is too
soft or room
temperature
is warm
Tilt of the
knife is too

embedded in
paraffin wax,
decalcification
is impractical;
use base
sledge
microtome
with a wedge
knife
Reduce the tilt

Tighten the
screw

Cut blocks into

small
fragments
Soften the
blocks in
detergent or
phenol
Breathe out or
blow gently on
the block and
knife to
breakup static
electricity, or
boil water in
the room to
increase the
humidity
Clean the
knife edge
Sharpen the
knife
Reduce the tilt
Sharpen the
knife

Re-embed in
filtered wax
Clean the
knife edge
with xylol
Reduce the tilt
Reduce the tilt
Sharpen the
knife
Cool paraffin
wax in ice
water
Increase the
tilt

the lower
part of the
section
during
cutting

small,
paraffin block
is therefore
compressed
against the
base of the
knife towards
the end of
stroke

FAULTS
Horizontal
or parallel
lines or
furrows
across the
section
(Chatters
) are seen,
forming
thin and
thick zones
Section cut
is
sometimes
thin,
sometimes
thick

REASON
Knife edge
vibrates due
to hardness
of tissue
Tilt of knife is
too great

REMEDY
Treat with
phenol during
processing or
collodionize
Reduce the tilt

Knife is blunt

Sharpen the
knife
Adjust the
knife

Knives
make a
hard
metallic
scraping or
ringing
sound on
backstroke,
when
section is
cut
Frozen
tissue
crumbles
and comes
off the
block holder
when cut
Frozen
tissue chips
into
fragments
when cut

Knife is not
clamped
properly
Tilt of knife of
block holder
is loose
Knife tilt is
too small that
block is
compressed
by bevel and
section is not
cut
Tilt of knife is
too slanted

Tighten
adjusting and
locking screws
Increase the
tilt

Knife blade is
too thin

Readjust the
angulation of
the knife
Take fresh
block treated
with phenol
during
processing
Change the
knife

Freezing is
not adequate

Refreeze the
tissue block

Tissue is
frozen too
hard

Warm the
tissue with
fingers

Tissue is too
hard