Histopath Instrumentation

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Honing

"sharpening" heel to toe

ultra-thin microtome

-.5 micra -broken plate glass -osmium tetroxide (fixed) -plastic embedded

Base sledge microtome

-2 movable pillars -RESIN EMBEDDED DECALCIFIED BONE

Cryostat or Cold microtome

-5 to -30 degree C 2-3 minutes 4u uses a medium = cryogel rotary microtome component enzyme study and Fluorescent Antibody staining technique most common rapid preparation

Freezing Microtome

-Quickett -Carbon dioxide passes from cylinder -hollow and perforemed around its perimeter -neuro and for sensitive tissue

Standard sliding microtome

-more dangerous -extremely hard tissue blocks (large intestine and small intestine)

Plane Wedge

100mm -hard and tough specimen frozen section

Biconcave

120 mm -both sides concave -recommened for paraffin

Coarse Honing and Honing Proper

2 types of Honing

Plane Concave

25mm one side flat concave paraffin and celloidin embedded tissue

Belgium Yellow Arkansas Fine Carborundum

3 honing types

Mechanical Honing

30 strokes each -driven by electric motor

Incubator

37 degree C overnight

Hot Plate

45-55 Degree C 30-45 mins

Wax oven

56-60 degree C 2 hours

Sliding Microtome

Celloidin sections

Belgium Yellow

For manual sharpening -usually gives best results

Freezing Microtome

Frozen sections

5-10 degree

Knife incline (clearance)

Rocking Microtome

Large Block of Paraffin

Hone

Natural Stone/hard grinding surface to sharpen the knife

Rotary Microtome

Paraffin embedded

Stropping

Polishing -removes burrs from honing Edge to last Toe to Heel

Plate Glass Hone

Uses Diamantine for Final Polishing

Xylene

after procedure wash or flush with _ _ _ _

27-32 degree

bevel angle

Mineral Oil

causes blister and destroys strope

Mineral Oil Clove Oil Xylene Liquid Paraffin Soapy water

common lubricants

76 x 25 mm 1.0-1.2 mm depth

dimension of a slide

Knife holder and knife

does the actual cutting

plane-wedge knife

each surface has 10-20 strokes

forceps and squirrel hair brush

for handling sections and removing folds

Padole Strope

from horse leather -firmly attached to a solid back to prevent sagging

Arkansas

gives more polishing effect

Machine Hone

glass disc/wheel driven by electric motor

Honing proper

grinding cutting edge at stone

Block Holder

holds tissue in position

40-120 double strokes

how many strokes do you need for stropping

20-30 times

how many times do you stroke for honing

vegetable oil or castor oil

leather strope is oiled with

8x3 inches

length of hone

Pawl, Ratchet Feed Wheel and adjustment screws

line up the tissue block

rotary microtome

minot -most common

Fine Carborundum

much coarser for badly nicked knives

plan concave knife

only the concave side is rubbed

15 degrees

optimum bevel angle

Rocking microtome

paldwell trefall -simplest - heavy base and 2 arms - 10-12 um

4u thick

paraffin wax block thickness

plate glass hone

powdered aluminum oxide (made into paste using water) -wider width no need to hold obliquely

Microtomy

process by which processed tissue most commonly being paraffin embedded is trimmed and cut uniformly thin slices or sections to facilitate studies under the microscope

detergent

reduce surface tension and allow the section to flatten out

Coarse Honing

removal of nicks on knife edge

brain or nervous tissue

sample utilized in hot plate

10 degrees below melting point of paraffin

temperature of water bath

Ultra-thin microtome

utilized for electron microscopy


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