Microtomy o This was invented by Minot in 1885 –
1886 to cut paraffin embedded tissues
• The process by which processed tissue, most and is currently the most common type
commonly a paraffin embedded tissue, is trimmed and used for both routine and research
cut into uniformly thin slices or "sections" to facilitate laboratories, especially for sectioning
studies under the microscope. paraffin-embedded tissues. Electrically
• Microtomy is a skilled process that requires precision driven rotary microtomes re also now
and hand - eye coordination combine with a delicate available and can be ideally used to
touch by the experience histotechnologist/cian produce ribbons for serial sections.
• Sliding microtome - for cutting celloidin embedded
Microtome sections.
o This was developed by Adams in 1789.
• The basic instrument that is capable of cutting a section o There are two types of this microtome.
at a o Base-Sledge Microtome, consists of two
• predetermined thickness by sliding the block into a movable pillars holding the adjustable
cutting tool, usually a steel knife, glass or diamond knife clamps, allowing the knife to be set
blade, which is fixed and attached to the machine. at an angle for cutting celloidin sections.
• Three Essential Parts: ▪ The Base-Sledge microtome is
o Block holder - where the tissue is held in favored in laboratories where
position very hard tissue or large blocks
o Knife Carrier and Knife - for actual cutting of are usually sectioned.
tissue sections. o Standard Sliding Microtome, the block
o Pawl, Ratchet Feed Wheel and Adjustment remains stationary while the knife is
Screws - to line up the tissue block in proper moved backward and forward during the
position with the knife, adjusting the proper process of sectioning.
thickness of the tissue for successive ▪ for cutting celloidin embedded
sections. tissue blocks
• Principle: A spring-balanced teeth or pawl is brought
into contact with, and turns a ratchet feed wheel • Freezing microtome -for cutting unembedded
connected to a micrometer screw, which is in turn frozen sections.
rotated, moving the tissue block at a predetermined o This was invented by Queckett in 1848.
distance towards the knife for cutting sections at The stage for block holder is hollow and
uniform thickness. perforated around its perimeter, attached
to a reinforced flexible lead pipe thru
which carbon dioxide passes from a
Kinds of Microtome cylinder.
o It is used to cut undehydrated thin to
• Rocking microtome – for cutting serial sections of
semi-thin sections of fresh, frozen
large blocks of paraffin embedded tissues
tissues, especially in instances when
o This was invented by Paldwell Trefall in
rapid diagnosis is required, when
1881, the simplest among the different
histological demonstration of fat is
types of microtomes.
needed, when certain neurological
o This consists of a heavy base and two
structures are to be studied, and when
arms the lower arm resting on pivots and
sensitive tissue constituents to be
a supporting column, and attached to the
studied are damaged or destroyed by
micrometer screw, at the base of which is
heat.
found the ratchet wheel with feed
mechanism.
• Cryostat or cold microtome – for cutting frozen
o A section is thereby cut as the tissue
sections
passes to the knife edge in a slightly
o The Cryostat is a refrigerated apparatus
curved plane, in 10-12 u thickness.
used for freezing the tissue into the block
o The Cambridge rocking microtome,
holder to the correct degree of hardness
available in two sizes, has been used to
that allows for easier and faster
cut small and large blocks of paraffin
sectioning.
tissues.
o It consists of a microtome, usually a
rotary microtome, kept inside a cold
• Rotary microtome - for cutting paraffin embedded
chamber which has been maintained at a
sections.
temperature between -5° to -30°C
(average is -20°C) by an adjustable
thermostat, capable of freezing fresh Stropping (TOE TO HEEL)
tissues within 2-3 minutes, and cutting
sections of 4 μ. • The process whereby the “burr” formed during honing
o The cryostat provides a means of is removed and cutting edge of the knife is polished.
preparing thin sections of fresh frozen
tissues especially for fluorescent
antibody staining techniques or Cutting Sections
histochemical enzyme studies.
o It is most commonly used for rapid • Sectioning is a process whereby tissues are cut into
preparation of urgent tissue biopsies for uniformly thin slices or "sections" with the aid of a
intraoperative diagnosis. microtome, to facilitate the studies under the
microscope.
• Ultrathin microtome - for cutting sections for
Electron Microscopy.
o An ultrathin microtome equipped with a Three general types of tissue sections
glass or gem grade diamond knife is used
to cut very thin sections (typically 60 to • PARAFFIN SECTIONS - for paraffin embedded tissue
100 nanometer) of tissue embedded in blocks which may be cut by rocking and rotary
epoxy resin. microtome.
o Sections are stained with an aqueous • CELLOIDIN SECTIONS - for celloidin embedded
solution of an appropriate heavy metal tissues which are usually cut by means of the sliding
salt and examined with a transmission microtome.
electron microscope (TEM). • FROZEN SECTIONS - which may be cut from tissues
that have been fixed and frozen with CO2 or for fresh
or fixed tissues frozen with the cryostat.
Factors to considered in sectioning
• Cutting depend upon the type of the tissue
• The size of the block
• The model or the type of the microtome
• *** sections usually 4-6 u in thickness for routine
histologic procedure
• **** The knife is usually tilted at 0-15° angulation on a
microtome
*”Bevel Angle" - The angle formed between the cutting edges normally about 27°
to 32°.
Knives for EM
• Glass knives
o Used for trimming and semi-thin sectioning of
tissue block
• Diamond knives
o Used to cut any type of resin block
o Brittle and expensive, but very durable
Honing (HEEL TO TOE)
• remove blemishes, grinding the cutting edge of the
knife on a stone (carborundum)
• Types of hones
o Belgium yellow – for manual sharpening (best
result)
o Arkansas – gives more polishing effect
o Fine carborundum – much coarser; Used only
for badly nicked knives