Freezing microtome_ cryostat.pptx
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1. To avoid drying, the tissue should be kept in OCT compound or freezing medium. 2. Tissues can be fixed with formalin. 3. Paraffin or celloidin is used as embedding media 4. Carbon dioxide gas is most commonly used with freezing microtome.
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This document provides information on microtomy, the process of cutting thin tissue sections for microscopic examination. It describes the basic components and types of microtomes used, including rotary, rocking, sliding and freezing microtomes. The steps of microtomy including tissue embedding, trimming blocks, mounting blocks, and cutting sections are outlined. Common problems like curled, thick/thin, or crumbling sections are discussed along with their causes and remedies. Requirements for producing high quality histological sections are also listed.
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This lecture helps to Define microtomy and microtomes, List the different types of them and Describe their functions.
Microtomy
Microtomy is the process of cutting thin tissue sections for microscopic examination. It involves using a microtome, a mechanical device that cuts very thin and uniform slices. There are various types of microtomes classified based on their cutting mechanism, such as manual microtomes like sliding, rotary, and cryostat microtomes, and automatic microtomes. Microtomes can also be classified based on the material of the knife edge used such as steel, glass, diamond, and sapphire knives. Proper sample preparation and microtome maintenance are important to obtain high quality sections without artifacts.
Microtome
Microtomy is the cutting of thin sections of objects for microscopic examination using a microtome. A microtome uses a mechanical device to precisely cut extremely thin, uniform sections, overcoming limitations of free-hand sectioning. There are two main types of microtomes: those where the block remains stationary and the knife moves, and those where the block moves across a stationary knife. Sledge microtomes are used to cut large samples embedded in paraffin, allowing sections from 1-60 μm to be cut.
Microtomy
This document discusses microtomy, which is the process of sectioning tissue samples for microscopic examination. It describes different types of microtomes used to cut thin, uniform slices of tissue, including rocking, rotary, sledge, sliding, and freezing microtomes. The mechanisms and uses of each microtome type are explained. Additionally, details are provided about microtome knives, angles, sharpening techniques, and types of abrasives used to maintain a sharp cutting edge for sectioning tissues.
Microtomy
This document discusses the process of microtomy, which involves preparing tissue samples for microscopic examination through sectioning. Key steps include:
1) Fixing tissue samples in formalin to preserve structure, then dehydrating them through a series of alcohol baths and clearing them in xylene.
2) Embedding tissue samples in paraffin wax, allowing it to solidify into blocks.
3) Sectioning the paraffin blocks with a microtome into thin slices, mounting them onto slides, and staining for examination.
4) Important considerations for microtomy include knife selection and maintenance, proper trimming and section thickness, and techniques for difficult tissues.
4. MICROTOMY.pptx
Microtomy is a method for the preparation of thin sections for materials such as bones, minerals and teeth, and an alternative to electropolishing and ion milling. Microtome sections can be made thin enough to section a human hair across its breadth, with section thickness between 50 nm and 100 μm
Microtomy and Paraffin Section Preparation ( PDFDrive ).pdf
The document provides guidance on proper microtomy and paraffin section preparation techniques. It stresses the importance of adequate fixation and processing of tissue specimens. Improper fixation and processing can result in tissue that is difficult to section. The document also discusses techniques for sectioning difficult blocks, optimizing the blade clearance angle, maximizing blade life, properly orienting specimens, considering factors that influence section thickness, and ensuring blocks are adequately chilled before sectioning.
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Freezing microtome_ cryostat.pptx
- 2. Introduction Microtomy Means by which the tissue comes in contact with the cutting tool (knife or blade) such that tissue is sectioned and attached to a surface for microscopic examination.
- 3. Introduction - 2 MICROTOME It is a mechanical device used for cutting uniform slices of tissue sections
- 4. Introduction - 2 MICROTOME It is a mechanical device used for cutting uniform slices of tissue sections
- 5. Types of Microtome There are several types of microtome, each designed for a specific purpose, although many have multifunctional roles.
- 6. Rotary microtome Rotation of a fine advancing hand-wheel by 360° degrees, moving the specimen vertically past the cutting surface and returning it to the starting position. Most common type
- 7. Types of rotary microtome Manual rotary microtome Completely manipulated by the operator
- 8. Types of rotary microtome Semi automated rotary microtome One motor to advance either fine or course hand wheel
- 9. Types of rotary microtome Fully automated rotary microtome Two motors that drive both the fine and the course advance hand-wheel
- 10. Types of Microtome Base sledge microtome - Specimen is stationary - Knife slides across the top of the specimen during sectioning - Used for large blocks, hard tissues, or whole mounts, - Neuro and ophthalmic pathology.
- 11. Types of Microtome Rotary Rocking microtome - One of the oldest design - Knife is fixed, the tissue block moves through an arc and strikes against the knife - Disadvantages: Size of the blocks that can be cut is limited - Sections are cut in a curved plane ✓ Cheap, reliable, simple to use ✓ Requires minimal maintenance
- 12. Types of Microtome Sliding Microtome - The knife is stationary - Specimen slides under the blade during sectioning. - This microtome was developed for use with celloidin-embedded tissue blocks.
- 13. Types of Microtome Ultra Microtome - Exclusively for electron microscopy - Ultrathin sections upto 10nm - Glass/ diamond/sapphire knives - Block is brought close to the knife edge under a microscope - As each section is cut, it is floated on a water bath adjacent to the knife
- 14. Types of Microtome Freezing microtome Consists of a fixed stage over which the knife moves Carbon dioxide gas is used to freeze tissue It absorbs large heat and rapidly freezes the tissue
- 15. Types of Microtome Vibrating microtome - High speed vibration is used - Designed to cut tissues which have not been fixed or frozen - Sections are thicker - Tissue is immersed in water - Enzyme histochemistry and ultrastructure histochemistry
- 16. Types of Microtome Cryostat - Refrigerated cabinet with a microtome housed in a deep freeze cabinet - - 15 to -30 degrees C - Good, thin high quality frozen sections - Most of the cryostats use a modified rotary microtome
- 17. PRINCIPLE of FROZEN SECTION ● When a tissue is frozen, the water within the tissue turns to ice and in this state the tissue is firm, with the ice acting as the embedding medium.
- 18. cryostat
- 20. ● Access to the chamber is via a sliding window. ● Working temperature : 0 to -35 degree Celsius ● Rotary microtome controlled by an external hand wheel. ● Freezing shelf ● knife holder
- 22. HEAT EXTRACTOR
- 23. HAND WHEEL
- 24. ANTI ROLL PLATE ❖ Prevent rolling or curling of tissue ❖ Glass plate supported on an aluminium frame ❖ Provides gap between underside of glass and upper surface of knife.
- 25. Anti roll plate
- 26. TEMPERATURE SETTINGS ● Digital display ● For most tissues: -15 to -23 degree Celsius is used ● For tissues with more fat: Colder temperature ● Temperature log maintained ● Defrosting done daily ● Rapid freezing to be done to reduce freeze artefacts
- 27. CHUCKS
- 38. Thank You
- 39. 1. To avoid drying, the tissue should be kept in ...................... 2. Tissues can be fixed with ...................... 3. ...................... or ...................... is used as embedding media 4. ...................... gas is most commonly used with freezing microtome