1 / 37

MICROSCOPY AND STAINING

MICROSCOPY AND STAINING. Leeuwenhoek’s Investigation. Metric Units. Light Properties. Wavelength. http://eyetricks.com/0101.htm. Light Properties. Reflection Transmission. Light Microscopy Types. Compound Bright Field. Light Properties. Resolution. Wavelength/Resolution Interaction.

ketan
Télécharger la présentation

MICROSCOPY AND STAINING

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. MICROSCOPY AND STAINING

  2. Leeuwenhoek’s Investigation

  3. Metric Units

  4. Light Properties • Wavelength http://eyetricks.com/0101.htm

  5. Light Properties • Reflection • Transmission

  6. Light Microscopy Types • Compound Bright Field

  7. Light Properties • Resolution

  8. Wavelength/Resolution Interaction

  9. Light Properties • Absorption • Refraction http://interactagram.com/physics/optics/refraction/

  10. Effect of Immersion Oil

  11. Disappearing Diamond

  12. Diffraction

  13. Diffraction

  14. Microscopy — Dark Field

  15. Microscopy — Phase Contrast • Dual Beam

  16. Microscopy — DIC (Nomarsky) • Differential Interference Contrast

  17. Microscopy — Fluorescence

  18. Microscopy — Confocal • Confocal

  19. Microscopy Imaging • Digital

  20. Same Image – Different Techniques

  21. Electron Microscopy • Transmission (TEM or EM) • Scanning (SEM) • Scanning Tunneling (STM) • Atomic Force (AFM) • Scanning Acoustic (SAM) http://micro.magnet.fsu.edu/primer/virtual/virtual.html *animation

  22. Freeze - fracture

  23. SEM Shadow Casting

  24. SEM freeze-etch

  25. Electron Microscopy Images

  26. Electron Microscopy Images

  27. STM

  28. AFM

  29. Microscopy Techniques • Wet Mounts • Smears • Drop

  30. Simple Stain • Provide basic contrast for light compound microscope • Anionic = acidic dyes: • eosin • picric acid • Cationic = basic dyes: • methylene blue • crystal violet • safranin • malachite green

  31. Differential Stain

  32. Differential Stain

  33. 1. Gram Stain Crystal violet stain Iodine mordant 95% ethanol decolorizer or ethanol/acetone Safranincounterstain

  34. 2. Acid Fast Stain (Ziehl-Neelson) • Paul Ehrlich in 1882; • Detecting tuberculosis and • leprosy – causing organisms. Carbolfuchsin stain 3% HCldeclorizer in 95% ethanol Methylene Blue counterstain

  35. 3. Special Stain: Negative Stain • Cell size • Cell shape • Indian Ink or acidic dyes • for background • Crystal Violet or Methylene • Blue for counterstain

  36. 4. Special Stain: Flagellar Stain • Silver; • Very difficult and time consuming;

  37. 5. Special Stain: Endospore Stain (Schaeffer-Fulton) Heat-fix Malachite Green stain Steam for 5 min to make the endo- spore wall permeable Rinse in water for 30 sec Decolor Safranincounterstain

More Related