Histopath Instrumentation
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
