I have read that different optical imaging techniques such as such as wide-field microscopy, confocal microscopy or STED microscopy can theoretically achieve a different spatial resolution.

However, I was only able to find information about the STED microscopy's spatial resolution (5.8 nm) on Wikipedia.

Does anybody know of any references or recommended reading (preferably free) where I can learn more about different optical microscopy methods and how the cope with the diffraction limit?

  • $\begingroup$ Super-resolution light microscopy will give you a spatial resolution on the order of FRET interaction distance (~10 nm) which is phenomenal considering the threshold was thought to be the limited by the diffraction limit. Conventional optical microscopy techniques can typically resolve down to ~200 nm. $\endgroup$ – user560 Apr 2 '12 at 0:57

The theoretical resolution for an optical microscope depends on the wavelength used, but is close to 0.22 um (220 nm). Confocal microscopy does not greatly improve axial resolution; it dramatically improves the in-plane resolution (x and y directions, by excluding extraneous light). Theres not a lot to do to "cope" with that using those techniques. If you need more resolution, you can use the super-resolution techniques you mention, or the good old TEM.

There is some more information on the Nikon website: http://www.microscopyu.com/articles/formulas/formulasresolution.html

and on Olympus website: http://www.olympusconfocal.com/theory/resolutionintro.html


You could also have a look at Leica Microsystems' Science Lab. You will find lots of articles and tutorials on the different microscopy methods, ranging from basic microscopy knowledge to specific know-how, including latest information on STED: www.leica-microsystems.com/science-lab

For example: gCW-STED Microscopy: When the Arrival Time of a Photon Matters http://www.leica-microsystems.com/science-lab/gcw-sted-microscopy-when-the-arrival-time-of-a-photon-matters/

From Molecules to Tissues – Optical Tools for Cancer Research http://www.leica-microsystems.com/science-lab/from-molecules-to-tissues-optical-tools-for-cancer-research/

(Disclosure: I have been involved in the launch of Leica Science Lab)


A couple of review articles you could read up on.

1) Leung BO, Chou KC. Review of super-resolution fluorescence microscopy for biology. Appl Spectrosc. 2011 Sep;65(9):967-80.

2) Huang B, Bates M, Zhuang X. Super-resolution fluorescence microscopy. Annu Rev Biochem. 2009;78:993-1016.

  • 1
    $\begingroup$ Might this not be better as a comment? :) $\endgroup$ – Rory M Apr 4 '12 at 19:46
  • $\begingroup$ Perhaps, but roseck did ask for some references to read, so I provided some appropriate articles. $\endgroup$ – user560 Apr 4 '12 at 19:47

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