Microtomy Flashcards
What is microtomy?
The process of preparing thin slices of biological tissue for microscopic examination of internal structure post embedding.
What is a microtome?
An instrument used to cut biological tissues into very thin, transparent slices for microscopic examination.
List the types of microtomes.
- Retracting
- Vibrating
- Rotary
- Sliding
- Freezing
- Ultramicrotome
Which microtome is most commonly used for sectioning paraffin embedded material?
Rotary microtome.
What are the main components of a microtome?
- Base
- Knife attachment and knife
- Material or tissue holder
How does a rotary microtome operate?
Most operate with a screw feed or by computerized motor, where the block moves up and down, advancing a preset number of micrometers with each revolution of the wheel.
What temperature should sections be floated on a floatation bath?
Approximately 46°C or 10°C below the melting point of the paraffin wax used.
When drying tissue sections, how should they be positioned?
Standing up to let water run down from underneath sections.
What is the recommended drying temperature for slides used in IHC?
Typically 40-50°C for 1 hour.
What factors affect the actual tissue section thickness in a microtome?
- Condition of the microtome
- Quality of knife edge
- Technologist skill
What is the primary maintenance requirement for microtomes?
Lubrication is essential to prevent friction and ensure smooth operation.
What are the advantages of using a freezing microtome?
- May be used to cut paraffin sections
- More portable and less expensive than the cryostat.
What are the disadvantages of a freezing microtome?
- Bulky and dangerous
- Parts must be securely attached
- CO2 bursts may transmit airborne diseases.
What materials are commonly used to make microtome knives?
- Glass
- Diamond
- Sapphire
- Disposable blades made of austenitic stainless or carbon steel.
What is the significance of knife edge quality in microtomy?
A good quality sharp knife can yield satisfactory sections even from poorly fixed tissue.
What are the four profiles of microtome knives?
- Profile A: strongly planoconcave or biconcave
- Profile B: planoconcave
- Profile C: wedge shaped
- Profile D: plane shaped
What is the recommended clearance angle for paraffin sectioning?
3° to 8°.
What are the consequences of an incorrect clearance angle?
- Too obtuse: knife scrapes instead of cuts.
- Too acute: paraffin block is crushed.
What is the bevel angle on a microtome knife?
Formed by upper and lower facets of the knife at 27° to 32°.
What is the purpose of a micrometer setting in a rotary microtome?
To determine the thickness of the tissue section.
What should be done with used knives?
Dispose of in a puncture-proof sharps container that is incinerated.
True or False: Xylene can be used to clean a microtome knife.
False.
What is the main advantage of automated microtomes?
They allow both hands to be free to manipulate ribbons.
What is a disadvantage of the freezing microtome in terms of safety?
The CO2 cylinder must be properly secured to prevent accidents.