1. Microscopes

2. Micrographia Robert HookeRobert Hooke 16651665 First report of cell structureFirst report of cell structure corkcork First illustrated book on microscopy

3. Binocular Microscope

4. Early Microscope Early compound microscopes were beautifully crafted but had severe chromatic distortion

5. Outline Units of Measurement Microscopes: features & uses –Light –Electron

6. Units of Measurement

7. Light Microscopes Brightlight Darkfield Phase Contrast Differential Interference Contrast Fluorescence Confocal

8. Parts of the Microscope

9. Changes in light as it passes through a cell

10. Bright Light Microscopy FEATURES structures > 0.2 um specimen dark against bright background cheap and easy USES Stained samples Simple morphology Count cells AKA Bright field microscopy

11. Stained bacteria

12. Shape is easier to see after staining

13. Granules & Capsules

14. Darkfield Microscopy FEATURES Special light source for indirect reflected light –condenser disk object bright against black background improved resolution USES living organisms fragile organism small microbes –Treponema pallidum

15. Flagella can be seen by Dark field microscopy

16. Dark field Microscopy Click on image to start video Light reflected Objects bright against dark background Slightly improved resolution

17. Phase Contrast Microscopy FEATURES Special condenser Special objective lens Uses diffracted light No stains USES Living Structures Motility Internal cell Structures

18. Phase Light

19. Effect of edges on phase light

20. Bacteria by Phase Contrast

21. Phase Contrast Microscopy Live cells internal organelles visible Dynamic processes Click on image to show Video

22. Differential Interference Contrast Microscopy FEATURES Two Light Beams Sample appears coloured USES 3D images First developed by Nomarski in 1952

23. World’s Largest Bacterium Visible to naked eye In gut of Surgeon fish Eats alga

24. Procloron under DIG Microscopy

25. Procloron dividing

26. Four Types of Light Microscopy BrightfieldPhase contrast NomarskiDarkfield

27. Fluorescence Microscopy FEATURES UV light source Special lenses objects fluorscent against black background USES Immunofluorescent labelled antibodies rapid identification

28. Light path in a flourescent Scope

29. Fluorecent cells

30. Fluorescent Molecules

31. Double fluorescent labelling

32. Confocal Microscopy FEATURES Laser light multiple images computer generated USES 2D and 3D of fine structural detail

33. Confocal Microscopy

34. Fluorescence Microscopy ConventionalConfocal

35. 3D Reconstruction Via Confocal Microscopy

36. Electron Microscopy Transmission Electron Microscopy Scanning Electron Microscopy

37. Lens in Microscopes

38. Plasma Membrane: Glycocalyx Glycocalyx of plasma membrane of a T lymphocyte stained with ruthenium red

39. Scanning EM

40. Bacteria on Skin

41. Flagella & Pili

42. Scanning EM

44. TEM SEM DIC Comparison of EM methods