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2. Microtomy Bio Techniques.pptx

1 de Apr de 2023
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2. Microtomy Bio Techniques.pptx

  1. Microtomy Biological Techniques
  2. What is microtomy? Introduction
  3.  Thickness of the sections is predetermined at regular distance on the microtome  The sections are attached to a surface like glass slides for staining  Tissues like muscles, bones, hair are cut of thickness from 50 nm and 100 µm  Thickness of the sections may be fixed according to the need of the study and material involved  The stained sections can be studied by light or electron microscopy Introduction
  4. Advanced Analytical Techniques Microtomy- Types of microtomy
  5. Types of microtomy  Main types include ◦ Rotary microtome ◦ Cryotome ◦ Ultra microtome ◦ Laser microtome  Use depends on the material under study and the nature of the work  Simple studies involve rotary microtome while detailed studies may involve ultra microtome, etc.  Similarly cryomicrotome involves sectioning of the frozen tissues.
  6.  The most common type of the microtome  Rotary action involves the sectioning process at predetermined thickness on every rotation of the flywheel  Blade is fixed at horizontal position  The sample holder moves the sample ahead by the fixed distance for cutting  Flywheel of the instrument may be automatic or manual  Section thickness may vary from 0.5 µm to 60 µm Types of microtomy
  7. Types of microtomy  Used for making extremely thin sections (40 nm to 500 nm)  Used mostly for biological samples but samples may also be processed  Linear thermal expansion moves the specimen at regular interval  The sections are studied in transmission or scanning electron microscopy  The cut sections are floated on the top of a liquid  These are then mounted on a copper, nickel, gold, or other metal grid
  8. Advanced Analytical Techniques Microtomy- Method (fixation and processing)
  9.  It consists of following main steps  Fixation  Processing  Dehydration  Clearing  Embedding  Section cutting  Staining  Deparaffinization Method (fixation and processing)
  10. Fixation Method (fixation and processing)
  11.  Commonly used fixatives are alcohol, formalin, glutaraldehyde, etc.  Factor affecting fixation are temperature, change in pH, penetration of the fixative, volume, time, etc.  The lowest concentration of the fixative is preferred than the higher one  10% formalin or 2.5% glutaraldehyde is used Method (fixation and processing)
  12. Processing Method (fixation and processing)
  13. Advanced Analytical Techniques Microtomy- Method (embedding)
  14. Embedding Method (embedding)
  15. Method (embedding)
  16. Method (embedding)
  17. Advanced Analytical Techniques Microtomy- Method (sectioning and staining)
  18. Sectioning Method (sectioning and staining)
  19. Sectioning Method (sectioning and staining)
  20. Staining Method (sectioning and staining)
  21. Staining Method (sectioning and staining)
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