TRIMMING OF SECTIONS 4.

Diamond-edge Knife—recommended
for ultrathin-sectioning microtome used
Once the wax has solidified, the wax for electron microscopy
block is removed from the mold, and the
excess wax is cut off from the block to
expose the tissue surface in preparation
for actual cutting .

The sides, top, and bottom of the tissue

block are trimmed until perfectly level
and all sides are parallel, almost to the
edge of the tissue, until the entire tissue
surface has been partly exposed.

MICROTOME KNIVES—used for trimming BEVEL ANGLE—angle formed between

and section-cutting the cutting edges, normally about 27° to
Available in 4 basic types: 32° is maintained for each knife by means
of a slideon knife back, a spring-loaded
1. Plane-Concave Knife—flat side is semi-circular metal sheet slipped on to
recommended for cutting celloidin the knife to hold the cutting edge at a
embedded sections on a sliding constant, correct angle during the
microtome; concave side is used to cut process of honing and stropping
paraffin embedded sections on
basesledge, rotary or rocking microtome

2. Biconcave Knife—used for cutting

paraffin embedded sections on a rotary
microtome

3. Plane-Wedge Knife—recommended for

frozen sections, or hard specimens
embedded in paraffin blocks, using a HONING AND STROPPING --Badly
base sledge or sliding microtome nicked knives with blunted ends have to
undergo sharpening in order to ensure
optimum sectioning of tissue blocks and 1. Block holder—where the tissue is held
prevent gross irregularities on the tissue in position
sections
2. Knife carrier and Knife—for actua
cutting of tissue sections

Sharpening involves 2 stages: 3. Pawl, Ratchet feed wheel and

Adjustment Screws—to line up the tissue
1. HONING—involves the removal of
block in proper position with the knife, for
gross nicks on the knife edge (coarse
adjusting the proper thickness of the
honing) to remove blemishes, and
tissue
grinding the cutting edge of the knife on a
stone (honing proper) to acquire an even 4. Principle: a spring-balanced pawl is
edge brought into contact with, and turns a
ratchet feed wheel connected to a
- Types of hones: Belgium Yellow,
micrometer screw, which is in turn
Arkansas, Fine Carborundum
rotated, moving the tissue block at a
- Knife edge first, with a “heel to toe” predetermined distance towards the knife
direction for cutting sections at uniform thick less

2. STROPPING—involves the removal of

the "burr" that have been formed during
5 kinds of microtome:
honing, and the final polishing of the knife
edge 1. ROCKING MICROTOME—for cutting
serial sections of large blocks of paraffin
- Paddle strop made up of horseleather
embedded tissues; 10-12 µ thick
- Knife edge last with a "toe to heel”
2. ROTARY MICROTOME—for cutting
direction
paraffin embedded sections; 4-6 µ thick;
most commonIy used for routine and
research laboratories
CUTTING OF SECTIONS TRIMMING OF
SECTIONS 3. SLIDING MICROTOME—for cutting
celloidin sections; 7-9 µ thick; 2 models:
Sectioning—process whereby tissues are
Base Sledge and Standard Sliding
cut into uniformly thin slices or "sections"
Microtome
with the aid of a microtome, to facilitate
microscopic study 4. FREEZING MICROTOME–for cutting
unembedded frozen sections; 10-15 µ
thick
3 essential parts of a microtome:
- Used to cut tissues in the frozen state, angle between the cutting facet and the
especially instances when rapid tissue block
diagnosis is required, when
- Sections form ribbons, complete
histochemical studies are needed (e.g.,
ribbons are picked up at once with a
fat, enzyme) and when tissue
camel hair brush, forceps, or fingers
constituents to be studied are damaged
by heat - Sections are then floated out on a water
bath set at 45-50°C (app. 6-10°C lower
than the melting point of the wax used for
• CRYOSTAT or COLD embedding the tissue), folds and creases
MICROTOME—apparatus capable may be removed by stretching the
of freezing the tissue into the sections gently with a pair of dissecting
blockholder to the correct degree needles
of hardness to facilitate easier and
- Sections selected for staining are picked
faster sectioning
up onto a clean glass slide in a vertical
-Consists of a microtome, kept inside a position; slide is immersed in the water
cold chamber, maintained at a bath in a near vertical position as close as
temperature of -5 to -30°C (average - possible to the section, when the slide
20°C) by an adjustable thermostat, touches the tissue section, it is lifted
capable of freezing fresh tissues within 2- vertically out of the water and drained—
3 minutes FLOATING-OUT or FISHING-OUT

- Tissue sections are then placed in a

paraffin oven (maintained at a
5. ULTRATHIN SECTIONING
temperature 2-5°C above the melting
MICROTOME—for cutting sections used
point of paraffin wax) to dry
in electron microscopy; 0.5 µ thick
2. CELLOIDIN SECTIONS—celloidin
Types of Tissue Sections:
embedded tissue blocks which may be
1. PARAFFIN SECTIONS—paraffin cut by sliding microtome, sections are cut
embedded tissue blocks which maybe between 7-9 µ thick
cut by rocking and rotary microtome
3. FROZEN SECTIONS—may be cut from
- Sections are cut between 4-6 µ thick for fresh or fixed tissues frozen with CO2,
routine procedures and to cut in a freezing microtome or
Cryostat.
- Knife is usually tilted at 0-15° angulation
on a microtome to allow a clearance