Part of the book series: Neuromethods ((NM,volume 86))

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Abstract

Galileo Galilei invented the first microscope “occhiolino,” by combining a concave and a convex lens in 1600s. Robert Hooke and Anton van Leeuwenhoek later modified it to look at living things. Since then, light microscopy has gained immense popularity and has been pushing the limits of optical technology. The race to improve the power of seeing continued by introduction of new techniques, more and more powerful lenses, better optical corrections, stronger light sources, higher-sensitivity detectors, and assembly of cutting-edge systems. However, the wish for a better peek at the cellular world hit a wall in the 1990s, already envisioned by Ernst Abbe in 1873. Diffraction, which enables an image to be formed in the first place, is also a barrier that obscures the finer details of cells and biomolecules. This chapter presents a discussion of resolution, briefly introducing some of the fundamental concepts such as diffraction and contrast.

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References

  1. Karlsson RC (2009) Milestone 1: The beginning. Nature Cell Biology 11:S6. doi:10.1038/ncb1938

  2. Hooke R (1665) Micrographia: or, some physiological descriptions of minute bodies made by magnifying glasses. With observations and inquiries thereupon. John Martyn and James Allestry, London

    Google Scholar 

  3. van Leeuwenhoek A (1977) The select works of anthony van leeuwenhoek: containing his microscopical discoveries in many of the works of nature. Arno Press, New York

    Google Scholar 

  4. Chalfie M, Tu Y, Euskirchen G et al (1994) Green fluorescent protein as a marker for gene expression. Science 263:802–805

    Article  CAS  PubMed  Google Scholar 

  5. Tsien RY (1989) Fluorescent probes of cell signaling. Annu Rev Neurosci 12:227–253. doi:10.1146/annurev.ne.12.030189.001303

    Article  CAS  PubMed  Google Scholar 

  6. Murphy DB, Davidson MW (2012) Fundamentals of light microscopy and electronic imaging, 2nd edn. Wiley-Blackwell, Hoboken

    Book  Google Scholar 

  7. Rayleigh L (1896) XV. On the theory of optical images, with special reference to the microscope. Philos Mag 42:167–195. doi:10.1080/14786449608620902

    Article  Google Scholar 

  8. Pawley JB (2006) Handbook of biological confocal microscopy, 3rd edn. Springer, New York

    Book  Google Scholar 

  9. Abbe E (1873) Beiträge zur Theorie des Mikroskops und der mikroskopischen Wahrnehmung. Archiv f mikrosk Anatomie 9:413–418. doi:10.1007/BF02956173

    Article  Google Scholar 

  10. Linfoot EH, Wolf E (1953) Diffraction images in systems with an annular aperture. Proc Phys Soc B 66:145–149. doi:10.1088/0370-1301/66/2/312

    Article  Google Scholar 

  11. Stelzer EHK (1998) Contrast, resolution, pixelation, dynamic range and signal-to-noise ratio: fundamental limits to resolution in fluorescence light microscopy. J Microsc 189:15–24

    Article  Google Scholar 

  12. Hell SW, Wichmann J (1994) Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. Opt Lett 19:780–782. doi:10.1364/OL.19.000780

    Article  CAS  PubMed  Google Scholar 

  13. Heintzmann R, Cremer CG (1999) Laterally modulated excitation microscopy: improvement of resolution by using a diffraction grating. In: Altshuler GB, Benaron DA, Ehrenberg B, et al. (eds) SPIE Proceedings. SPIE, pp 185–196

    Google Scholar 

  14. Gustafsson MGL (2005) Nonlinear structured-illumination microscopy: wide-field fluorescence imaging with theoretically unlimited resolution. Proc Natl Acad Sci U S A 102:13081–13086. doi:10.1073/pnas.0406877102

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  15. Hell S, Stelzer EHK (1992) Properties of a 4Pi confocal fluorescence microscope. J Opt Soc Am A 9:2159. doi:10.1364/JOSAA.9.002159

    Article  Google Scholar 

  16. Gustafsson MG, Agard DA, Sedat JW (1999) I5M: 3D widefield light microscopy with better than 100 nm axial resolution. J Microsc 195:10–16. doi:10.1046/j.1365-2818.1999.00576.x

    Article  CAS  PubMed  Google Scholar 

  17. Rust MJ, Bates M, Zhuang X (2006) Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat Methods 3:793–796. doi:10.1038/nmeth929

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  18. Betzig E, Patterson GH, Sougrat R et al (2006) Imaging intracellular fluorescent proteins at nanometer resolution. Science 313:1642–1645. doi:10.1126/science.1127344

    Article  CAS  PubMed  Google Scholar 

  19. Hess ST, Girirajan TPK, Mason MD (2006) Ultra-high resolution imaging by fluorescence photoactivation localization microscopy. Biophys J 91:4258–4272. doi:10.1529/biophysj.106.091116

    Article  CAS  PubMed Central  PubMed  Google Scholar 

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Saka, S.K. (2014). Light Microscopy and Resolution. In: Fornasiero, E., Rizzoli, S. (eds) Super-Resolution Microscopy Techniques in the Neurosciences. Neuromethods, vol 86. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-983-3_1

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  • DOI: https://doi.org/10.1007/978-1-62703-983-3_1

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-982-6

  • Online ISBN: 978-1-62703-983-3

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