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Principles of Confocal Microscopy

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Short presentation about optical principles of point scanning confocal microscopy. (5min presentation)

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Principles of Confocal Microscopy

  1. 1. THE PRINCIPLES OF POINT SCANNING CONFOCAL MICROSCOPY NICOLE SALGADO C. University College Dublin Science School MSc Imaging and Microscopy Paola Cognigni and Irene Miguel-Aliaga (2011)
  2. 2. WHAT IS ? POINT SCANNING CONFOCAL MICROSCOPY •Optical imaging technique used to increase optical resolution of a micrograph •Allows optical sectioning of a specimen •Patented by Minsky in 1957 PHOTOMULTIPLIER DETECTOR SCANNING HEAD ACQUISITION SOFTWARE NIKON Aퟣ CONFOCAL MICROSCOPE LASER SYSTEM Nikon Corporation (2014)
  3. 3. HOW DOES IT WORK? THE INSTRUMENT Modified from Tony B. Gines and Michael W. Davidson (2014)
  4. 4. HOW DOES IT WORK? THE OPTICAL PRINCIPLE Creative Commons, Rice University. (2014) Nathan S. Claxton , Thomas J. Fellers , Michael W. Davidson
  5. 5. WHY DO WE USE IT? ADVANTAGES OF CONFOCAL MICROSCOPY •Increased resolution •Optical sectioning •3D reconstruction Nathan S. Claxton , Thomas J. Fellers , Michael W. Davidson Thomas J. Fellers and Michael W. Davidson. (2009)
  6. 6. WHAT ARE THE PRECAUTIONS? •Scanning speed •Photobleaching •Settings of the Photomultiplier detector Brian Herman, Matthew J. Parry-Hill, Ian D. Johnson, and Michael W. Davidson (2014)
  7. 7. REFERENCES •Paola Cognigni and Irene Miguel-Aliaga (2011). Fly poo never looked so beautiful…. Available: http://blog.wellcome.ac.uk/2011/01/06/fly-poo/. Last accessed 30th Sept 2014 •Nikon Corporation (2014). Confocal Microscopes A1+/A1R+. Available: http://www.nikon.com/products/instruments/lineup/bioscience/confocal/singlephoton/a1/. Last accessed 30th Sept 2014. •Tony B. Gines and Michael W. Davidson (2014). LSM 700 Light Pathways in Spectral Imaging. Available: http://zeiss-campus.magnet.fsu.edu/tutorials/spectralimaging/lsm700/indexflash.html. Last accessed 1st Oct 2014. •Kenneth R. Spring, Thomas J. Fellers and Michael W. Davidson (2009). Confocal Microscope Scanning Systems. Available: http://www.olympusconfocal.com/theory/confocalscanningsystems.html. Last accessed 1st Oct 2014. •Australian Microscopy and Microanalysis Research Facility (2014). Confocal Microscopy . Available: http://www.ammrf.org.au/myscope/confocal/confocal/. Last accessed 2nd Oct 2014. •Mortimer Abramowitz and Michael W. Davidson (2012). Photomultiplier Tubes. Available: http://www.olympusmicro.com/primer/flash/photomultiplier/. Last accessed 1st Oct 2014. •Creative Commons, Rice University. (2014). *Extended Topic* Microscopy Enhanced by the Wave Characteristics of Light. Available: http://cnx.org/contents/0cc2b3c8-487e-49ea-925f-ff51cb2a0591@2. Last accessed Oct 1st 2014. •Nathan S. Claxton , Thomas J. Fellers , Michael W. Davidson. (). LASER SCANNING CONFOCAL MICROSCOPY. Available: http://www.olympusfluoview.com/theory/lscmintro.pdf. Last accessed 30th Sept 2014. •Thomas J. Fellers and Michael W. Davidson. (2009). Introduction to Confocal Microscopy. Available: http://www.olympusfluoview.com/theory/confocalintro.html. Last accessed 2nd Oct 2014. •Brian Herman, Matthew J. Parry-Hill, Ian D. Johnson, and Michael W. Davidson (2014). Photobleaching. Available: http://micro.magnet.fsu.edu/primer/java/fluorescence/photobleaching/. Last accessed 2nd October 2014.

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