Dr. S.

Manickam

 Understand

the principles of microtomy Describe the different types of microtomes and their specific purposes Describe the factors affecting section quality Describe the purpose of adhesives

 Knowledge

of equipment used Practical experience

Classified by the manner they are ground: Plane concave Plane concave (Very concave) Plane wedge Biconcave

 Sledge

microtome

 Celloidin

section with hard

Thin
Vibrates

tissues

 Frozen

section Paraffin wax section Rigid Suitable for sledge microtome

 Paraffin

wax sections Used in sledge & Rocking microtomes

Cutting edge should be straight line formed by intersection of 2 planes, the cutting facets

Angle between planes Bevel angle (27 - 32)

Assessed by seeing under microscope at 100X

Grinding, renews bevel, restores edge to straight line Belgium black vein Arkansas Carborundum Plate glass with abrasives like aluminium oxide

Measures 8 x 2 inches

 Polishes

already sharp edge Hide of rump of horse

Knife

sharpening machines

Facet angle is 35o for routine cutting Teflon reduces resistance during cutting; platinum gives edge of blade strength

Clamped into welldesigned blade holder

Mechanical devices for cutting thin uniform slices of tissue (sections) Coined by Charles Chevalier, who perfected it around 1825

5 Basic types: Rocking Rotary Sledge Sliding Freezing

 Knife

is fixed Block moves through arc & strikes against knife Adv: Small blocks, simple, long lifetime
Disadv: Used:

Lightness

Paraffin & Frozen sections

 Rotary

action of handwheel Adv: Heavier, ideal for serial sections, large blocks
Used:

Paraffin & celloidin sections

For very large tissue blocks Knife is fixed Block in steel carriage moves forwards & backwards

Adv: Heavy, no vibration Disadv: Slower

Used: Paraffin & Celloidin sections

 Knife

moves horizontally Block remains fixed

Used:

Celloidin sections

 Semi-thin

and ultrathin sections Resin-embedded specimens Glass or diamond knives Block or knife retraction between strokes

 Other

microtomes can be modified This gives best results

sections (3-5) of frozen tissue Rotary microtome in deep freeze cabinet

Thin

 1/8

inches on all sides of tissue Edges should be parallel Surface should be trimmed with microtome

 Block

in block holder Insert knife, check tilt Blocks and Knifes edges should be parallel Trim the surface Set thickness to 6 Cut sections and hold the free end with needle or forceps

 In

cutting paraffin wax section, each section sticks to the previous one by its edge

 If

every fifth or tenth section is taken for study

Cutting makes sections Compressed Creased It should be removed & flattened Float them on warm water Waterbath method Hot stage method Warm slide method

 Albumin

Starch

paste Chrome gelatin

Faults Section cut vertically

Reasons Knife edge is damaged

Section rolls up
Section crumbles on cutting Ribbon curves

Knife is blunt or tilted more

Knife blunt, wax is soft Block edges are not parallel to each other or to knife

Tissue embedded on ice & sections cut on freezing microtome

To demonstrate fat, lipids or special components For rapid preparation for Dx

Fixation: Formal saline, 60, 10 min Gum syrup 8 -10 min