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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?

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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. – user560 Apr 2 '12 at 0:57
up vote 7 down vote accepted

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:

and on Olympus website:

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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:

For example: gCW-STED Microscopy: When the Arrival Time of a Photon Matters

From Molecules to Tissues – Optical Tools for Cancer Research

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

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Thanks for the disclosure =D – Rory M Apr 4 '12 at 19:36

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.

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

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