Your SlideShare is downloading. ×
0
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy
Upcoming SlideShare
Loading in...5
×

Thanks for flagging this SlideShare!

Oops! An error has occurred.

×
Saving this for later? Get the SlideShare app to save on your phone or tablet. Read anywhere, anytime – even offline.
Text the download link to your phone
Standard text messaging rates apply

Microscopy dental applications /certified fixed orthodontic courses by Indian dental academy

1,320

Published on

Welcome to Indian Dental Academy …

Welcome to Indian Dental Academy
The Indian Dental Academy is the Leader in continuing dental education , training dentists in all aspects of dentistry and offering a wide range of dental certified courses in different formats.


Indian dental academy has a unique training program & curriculum that provides students with exceptional clinical skills and enabling them to return to their office with high level confidence and start treating patients

State of the art comprehensive training-Faculty of world wide repute &Very affordable.

Published in: Education
0 Comments
3 Likes
Statistics
Notes
  • Be the first to comment

No Downloads
Views
Total Views
1,320
On Slideshare
0
From Embeds
0
Number of Embeds
1
Actions
Shares
0
Downloads
0
Comments
0
Likes
3
Embeds 0
No embeds

Report content
Flagged as inappropriate Flag as inappropriate
Flag as inappropriate

Select your reason for flagging this presentation as inappropriate.

Cancel
No notes for slide

Transcript

  1. MICROSCOPY Dental applications INDIAN DENTAL ACADEMYLeader in Continuing Dental Education www.indiandentalacademy.com
  2. WHY DO WE NEED MICROSCOPES
  3. CONTENTS….• History of microscopy• The basics concepts of optics• Optical phenomena of interest to microscopy• Microscopes in dental research - principles, specimen preparation and applications• Choice of microscopes for successful research• Microscopes in clinical dentistry• Conclusion
  4. A WALK DOWN MEMORY LANE….. Leeuwenhoek- Binnig and Rohrer - Simple microscope Scanning tunnelling Hillier- microscope Galileo - TEM Occhiolino 1609 1674 1938 1981 1590 1665 1931 1957 1983Jannsen – Ruska –Compound Electron microscope Binnig -microscope Atomic force microscope Robert Hooke - Micrographia Petran and Minsky – Confocal microscopy
  5. THE BASICS• MAGNIFICATION• IMAGE QUALITY• BASIC OPTICS AND OPTICALPHENOMENA
  6. MAGNIFICATION• ENLARGING SOMETHING ONLY IN APPEARANCE• OPTICAL MAGNIFICATION (Power) – Apparent size : Truesize…….at 25 cm distance• LINEAR AND ANGULAR MAGNIFICATIONS• HIGHER MAGNIFICATION LENSES GENERALLYHAVE HIGH RESOLUTION MAGNIFICATION (M) = M0 x Me• Useful magnification…• Depth of field ……• Field number and size….
  7. RESOLUTIONThe ability to differentiate between two closely positioned brightobjects. Resolving power line* Light microscope includes phase contrast and fluorescence microscopes. Electron microscope includes transmisson electron microscope.
  8. THE BASIC CONCEPTS OF OPTICS - OPTICAL PHENOMENA OF INTEREST TO MICROSCOPY REFLECTION Vs REFRACTIONrefraction reflection
  9. Diffraction…… light bending around an object Airy diffraction….
  10. PATHWAYS OF LIGHT…… EYEPIECE AND OBJECTIVE LENS – CONVEX LENSES Converging lens….. Focal point focal length POWER (diopters) = 1 / f
  11. The real The magnified image…. image.. objectobject f 2f 2f f IMAGE IMAGE
  12. Image formed by the microscope….. Eyepiece Objective Image of Object (X1) Object (X) Final image (X2) X1 – a real, inverted and bigger image X2 – a magnified and virtual image
  13. TOTAL INTERNAL REFLECTIONREFRACTION OF LIGHT AT A MEDIUMBOUNDARY,ENOUGH TO SEND IT BACKWARDS ANDCAUSE EFFECTIVE REFLECTION OF THE LIGHT Refraction at medium boundary
  14. THE EVANESCENT WAVE• EVANESCENT = “tending to vanish”• DECAYS EXPONENTIALLY WITH DISTANCE• RADIATION PRESSURE TO ILLUMINATE SMALL OBJECTS• Selective excitation of fluorophores
  15. FLUORESCENCE
  16. FOCUS , CONFOCAL AND PARFOCALFOCUS –• point of convergenceCONFOCAL - "having the same focus."PARFOCAL – “having the same focus in the entirerange of magnifications”
  17. Categorization of microscopy ….Illumination methods Imaging method Transmitted Reflected• visible light• laser• electrons• X-rays• non-illumination type
  18. MICROSCOPES IN DENTAL RESEARCH• LIGHT MICROSCOPE• PHASE CONTRAST MICROSCOPE• STEREOMICROSCOPE• TOOLMAKER’S MICROSCOPE• FLUORESCENCE MICROSCOPE• TOTAL INTERNAL REFLECTION MICROSCOPE• ELECTRON MICROSCOPE  scanning electron microscope • field emission scanning electron microscope  transmission electron microscope• CONFOCAL MICROSCOPE• SCANNING PROBE MICROSCOPES  atomic force microscope  scanning tunneling microscope• X-RAY MICROSCOPE
  19. LIGHT MICROSCOPE• The simplest form of microscope eyepiece• Types based on illumination – bright, dark, objective rheinberg, phase contrast, differential interference, Hoffman modulation, sample polarised, fluorescence stage condenser• 0.5 – 1mm thin specimen• Image formation due to source of illumination absorption of light by objects in the path
  20. • Objective lens – 4x, 10x, 40x,100x• Eyepiece – 10x• Resolution : 200 nm (Ernst Abbe’s resolution limitation)
  21. PHASE CONTRAST MICROSCOPE• Enhances contrast• Altering the Light Waves• The Invisible Can Be Seen• Preparation of Specimen
  22. STEREOMICROSCOPY• Cherubin d’Orleans – 1671• Wenham and Greenough -1890• Optical light microscope• Stereoscopic vision – spatial 3- dimensional images• Designs – Cycloptic (common main objective), Greenough.
  23. Illumination and other aspects Sources of illumination: – Episcopic and diascopic illumination – Fluorescence illumination – LED (Chambers and Nothnagle – Microscopy and microanalysis – 1997) Total magnification : 2x to 540x Resolution : 1.6 µm
  24. TOOLMAKER’S MICROSCOPE• Precision measuring instrument• Linear scales• Magnification…..• Illumination : – Brightfield and darkfield – Rheinberg lighting – Polarisation – Phase contrast
  25. Fluorescence microscope• Concept of fluorescence and epi-fluorescence.• The sample itself is the light source.• Fading - photobleaching•
  26. Specimen preparation• Fluorescent Dyes• Immunofluorescence
  27. Total internal reflection microscope• Main disadvantage of fluorescence microscope – background fluorescence• Evanescent wave• 200 nm depth• Cell adhesion
  28. ELECTRON MICROSCOPE• Development of the electron microscope• Energy of the electrons – 1 KeV to 1 MeV.• Types• Gold standard in dental research….?
  29. • TRANSMISSION ELECTRON MICROSCOPE – Max Knoll and Ernst Ruska(1931)- (Nobel prize 1986)•SCANNING ELECTRON MICROSCOPE – Zworikyn -1942 • USES – Topographical imaging – Morphology imaging – Substructure analysis – Crystallography – Chemical composition
  30. BASICPRINCIPLES Electron source (or) the gun – 5 – 50 KV energy electrons ELECTRON DENSE ELECTRON TRANSLUCENTELECTRON TRANSPARENT
  31. Electron optics • Electron beam trajectory – electrostatic lenses (in the electron gun) and magnetic lenses • Double deflection scanning (A.W. Crew – Science – 1961)
  32. ELECTRON – SPECIMEN INTERACTIONS• BULK SPECIMEN INTERACTIONS• THIN (FOIL) SPECIMEN INTERACTIONS
  33. SCANNNING ELECTRON MICROSCOPYMAGNIFICATION20 X TO 65000 XRESOLUTION upto5 to10 nm
  34. Specimen preparation for scanning electron microscopy Fixation of specimen Dehydrate and embed in resin Section and Stain Gold / Platinum sputtering(Weimer and Martin – 13th International Congress on Electron Microscopy – 1974)
  35. SPUTTER COATER • Why sputter coat ? • Thickness…. • Materials used and method…. • In all forms of SEM ??
  36. FIELD EMISSION SCANNING ELECTRON MICROSCOPY - Erwin Müller (1936)• Materials science and bio-sciences• metal tip in a low vacuum chamber• Emitter gun – fine wire cathode and concave anode with a hole in the centre• Variation of atomic orientation in emitter tip• Sputtering…. • Resolution 20 Å • Magnification 50,000 X
  37. TRANSMISSION ELECTRON MICROSCOPYRESOLUTION0.2 nm
  38. Specimen preparation for Transmission electron microscopy ( Auvert – Microscopy and Microanalysis – 2003) Method 1 • Thickness – 500 nm (or) lesser • Fix in plastic (or) isolate and study as solution. • Spread on a support grid coated with plastic. • Heavy metal salt solution -stains -"shadow" around the specimen . •Negative picture
  39. Method 2 Pre – thinning of samples to 100 – 150 µm Polishing Platinum deposition (standard replica technique) Replication for bulk samples
  40. CONFOCAL MICROSCOPY• “Confocal imaging” – Nipkow• First stage scanning confocal microscope – Minsky (1957)• Petran, Minsky, Amos and White – Confocal Raman microscope – 1986• Types -
  41. CONFOCAL PRINCIPLE • “Confocal” – “having the same focus” • Confocal pinhole • Disadvantage of the pinhole • Compensation…
  42. Fluorescence in confocal microscopy At focal point Screen with pinholeThe pinhole is the CONjugate to the FOCAL point of the lens
  43. Confocal microscope – working Photomultiplier detectorPinhole aperture Barrier filter Dichroic mirror Laser Pinhole aperture objective Specimen focal planes
  44. A modification….. The real time tandem scanning microscope (Nipkow disc scanning confocal microscopy) (Watson and Wilson – Journal of microscopy – February 2002)• Nipkow disc – rapid scanning• Advantage – laser / fluorescence / non coherent light (xenon arc) , direct and safe visualisation (Mc cabe and Hewlett-Applied optics – January 1998)• No unwanted back reflections
  45. Specimen preparation for confocal microscopy Fixation (30 minutes , paraformaldehyde) Fluorophore labelling Elevation of coverslip (Immunocytochemistry – Polak and Noordan) specimen specimen
  46. PROS AND CONS• Thin optical sections (0.5 – 1.5 • Harmful nature of highµm) intensity lasers• Image formation restricted to a •Laser lines – excitationwell defined plane wavelength• Improved contrast and definition • Zoom factor – low• Multi dimensional view scanning rate• 1.2 nm resolution • High cost
  47. ATOMIC FORCE MICROSCOPE (Scanning force microscope)• Binning, Quate and Gerber – 1986• Imaging at nanoscale• Abrasion, adhesion, etching, corrosion, friction and polishing
  48. Working principle • Probe in the end of a cantilever • Flexion of cantilever • Measurement of changes in bending of cantilever by measuring difference A-B ( by Hooke’s law)
  49. Modes of operation and other aspects• Working modes may be in air (or) vacuum• Tip – sample interaction – Contact mode – Non contact mode – Tapping mode – Lift mode• Non destructive imaging is possible (E.Meyer – Microscopy and science – 1992)• Resolutions : – Vertical : 0.1 Å – Lateral : 30 Å
  50. Pros and Cons• Three dimensional surface • Image size – area ofprofile only 100 µm X 100• No special treatment for the µmsample - 8” diameter and 0.5” • Slow scanning ratethick . (Zhong and Innis – Surface Science – 1993)• Ambient and liquidenvironment• Very good resolution
  51. SCANNING TUNNELING ELECTRON MICROSCOPY ( STEM)  Binning and Roehrer – 1981….(Nobel Prize -1986)  Scanning technique within a transmission arrangement  Substrates – extremely flat - gold / platinum coating  Operated in vacuum  Vertical resolution : 0.1Å, lateral resolution : 1Å  Chemical analysis, topography
  52. X - RAY MICROSCOPY• Kirkpatrick and Baez• Electromagnetic radiation in the X-ray band• Do not reflect (or) refract easily• Charged coupled device (or) a film to detect the X rays that pass through the specimen• Solid – liquid interface analysis (Kaukler et al – NASA materials division research journal – 1999)
  53. LOADS OF OPTIONS IS ALWAYS CONFUSING !!!!
  54. SUCCESSFUL RESEARCH.… CHOICE OF MICROSCOPES…….• WHAT DO YOU WANT TO SEE ?• WHAT IS YOUR SPECIMEN SIZE ?• WHAT RESOLUTION DO YOU NEED ?• HOW MUCH ARE YOU READY TO SPEND ??!!
  55. FEATURES LIGHT SEM TEM CONFOCAL MICROSCOPE MICROSCOPESPECIMEN 0.1 – 0.5 mm Not specific….. To be less than No specificationTHICKNESS Ideally upto 3 cm 500 nm thick specimenRESOLUTION 200 nm 5 – 10 nm 0.1 – 0.2 nm 1.2 nmMAXIMUM Upto 2000 x, fixed Can be increased Can be increased 60 xMAGNIFICATION magnification upto 60,000 x upto 70,000 xMAIN USES IN Topography Topography, Same as SEM Topography,DENTAL ARENA morphology, morphology, chemical analysis, substructure substructure analysisMAJOR PROS Easy availability, Very good The best Easiest specimen easy procedure, resolution resolution preparation, economic good resolutionMAIN CONS Resolution Gold sputtering, Very thin sample Laser beam, Slow limitation cost factor, only needed, only dead scanning rate, dead cells cells Cost factor Cost factor
  56. MAGNIFICATION AND MICROSCOPY IN CLINICAL PRACTICE A Quantum leap……!!!!
  57. FROM “LOUPES” TO “SCOPES”…..• A fascinating moment in dentistry !!!!........ A pinnacle of technology…..• First ever use of magnification and illumination in endodontics – MAGNIFYING LOUPES (surgical telescopes) WITH FIBREOPTIC HEADLAMP• 1953 – Carl Zeiss company – binocular operating microscope• 1981 – Dentiscope – first dental operating microscope• 1993 – surgical operating microscope• Microscope Assisted Precision Dentistry
  58. SURGICAL OPERATING MICROSCOPE …. ANATOMY• body tube• Eyepiece and binoculars• Magnification changers and power zoom changers• Objective lenses – focal lengths 100 – 400 mm
  59. THE TRIFECTA ….. Magnify, illuminate and instrument• SURGICAL TELESCOPES : 2 – 3.5 x magnification• AVAILABLE MAGNIFICATIONS…..• HOW MUCH MAGNIFICATION DO WE REALLY NEED….usable power..?
  60. BASIC INFORMATION RELATING MAGNIFICATION TO OTHER FACTORS…….• f objective lens o magnification factor - magnification – ↓magnification -↓field of view – field of view – ↓illumination o power eyepiece - magnification -↓field of view• f binoculars – magnification o magnification - ↓depth of field – ↓ field of view
  61. ILLUMINATION IN OPERATING MICROSCOPE  Light source  xenon halogen bulb  quartz halogen bulb  Path of light and the “stereoscopic effect”  The Beam splitter  “co axial” illumination – “GALILEAN OPTICS”
  62. MICRO INSTRUMENTS• Micro mirrors, explorers, pluggers, burnishers, scalpels• Ultrasonic instruments – CPR, BUC, CT tips
  63. DOCUMENTATION AND ACCESSORIES….. • Production of quality images / videos / slides • Photo and cine adapters • Handles and LCD screen
  64. Operating positions……
  65. Mandibular rightMandibular left
  66. Mandibular anteriorMaxillary anterior
  67. Maxillary rightMaxillary left
  68. APPLICATIONS……..IN RESTORATIVE DENTISTRY IN ENDODONTICS • Microfractures • Diagnosis – untreated canals • Locating hidden canals • Caries diagnosis – • Calcified canals microdentistry • Perforation repair • Broken instrument retrieval • Final examination • Surgical endodontics • Patient education
  69. To conclude…..• Knowledge of physics of microscopes - essential to choose microscopes for research• Clinical practice with operating microscopes ….not a fancy, but a necessity!
  70. THANK YOU !!

×