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Exploring the Microscopic World: A Beginner's Guide to Microscopy

Discover the fascinating world of microscopy, from light microscopes to electron microscopy, learn the differences between TEM and SEM, and explore the various applications with colorful examples. Keen eyes required!

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Exploring the Microscopic World: A Beginner's Guide to Microscopy

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  1. Microscopy Looking at the small stuff

  2. Microscopes • “micro” means : • Small • “scope” means: • Watch/ look • It was actually developed first as entertainment for the rich

  3. Light Microscope • Focuses visible light rays • Can be used on live or dead specimens • Dyes and fluorescent markers help for viewing • Magnify up to 1000X on the ones we have • Examples: • Compound light microscope • Confocal microscope

  4. Light Microscope

  5. Light Micrographs Plant magnified Human chromosomes 

  6. Confocal Microscope

  7. Confocal micrograph Spirogyra crassa Maximum projectiongreen: Actin. Rhodamin, Label: Phalloidin. Actinred: Chloroplasts. AutofluorescenceImage acquired with the Leica TCS SP2

  8. Electron Microscopy (EM) • Focus beam of electrons on the specimen • Specimens must be dead • Kinds • Transmission Electron Microscope (TEM) • Cross-section of the material • Most detailed of internal of cell • Scanning Electron Microscope (SEM) • Surface of the specimen • Shows texture, but less detail

  9. TEM

  10. TEM

  11. SEM

  12. SEM Ant Pollen Grains

  13. Red Blood Cells

  14. Compare cardiac myocytes Compare to TEM of myoctes on the wall. Maximum projection (overlay) of neonatal cardiac myocytes from rat.green: Myomesin, FITCred: Cadherin, Cy3blue: DNA (nuclei), DRAQ5Image acquired with the Leica TCS SL

  15. Keen eyes • Light microscopes will not show you the nucleus of a cell or something the size of a virus! • Confocal microscopy uses brightly colored markers • TEM is detailed, flat, and black and white • SEM is 3D, textured, and black and white

  16. Cited pictures • Pollen grains (1000x magnification). Paul Fuqua. 2003.unitedstreaming. 29 August 2006http://www.unitedstreaming.com/ • Blood cells, red; microscopic view. U.S. Army. 2006.unitedstreaming. 29 August 2006http://www.unitedstreaming.com/ • http://www-medlib.med.utah.edu/WebPath/HEMEHTML/HEME001.html • www.plymouth.ac.uk/upload/SCI/liverartery.jpg • http://www.confocal-microscopy.com/WebSite/pictures.nsf/(ALLIDs)/23C6531FD6B41E6BC1256A7D0053F8CA/$FILE/drosoegg_app_img_i_cl.jpg • http://www.confocal-microscopy.com/WebSite/pictures.nsf/(ALLIDs)/23C6531FD6B41E6BC1256A7D0053F8CA/$FILE/drosoegg_app_img_i_cl.jpg

  17. Cited Pictures • http://remf.dartmouth.edu/images/insectPart3SEM/image/31formicidae40(extra).jpg • http://mikes-machine.mine.nu/Mysterymoss/Micro3_DSCN6486.jpg • http://www.nims.go.jp/analysis/TEM.jpg • http://www.nims.go.jp/analysis/SEM.jpg • http://cdr.rfmh.org/NewFiles/Components/images/confocal%20microscope.jpg • http://131.229.114.77/microscopy/images/chromosomes2.jpg • http://www.biosci.ohio-state.edu/~plantbio/osu_pcmb/images/Courses/PB101/plant_cell_tem.jpg

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