Chapter 12: Nanoscale biological fluorescence imaging: breaking the diffraction barrier.
Biological imaging has been limited by the finite resolution of light microscopy. Recent developments in ultra-high-resolution microscopy methods, many of which are based on fluorescence, are breaking the diffraction barrier; it is becoming possible to image intracellular protein distributions with resolution of tens of nanometers or better. Fluorescence photoactivation localization microscopy (FPALM) is an example of such an ultra-high-resolution method which can image living or fixed cells with demonstrated lateral resolution of better than 20 nm. A detailed description of the methods involved in FPALM imaging of biological samples is presented here, accompanied by comparison with existing methods from the literature.
Gould
Travis J
TJ
Department of Physics and Astronomy and Institute for Molecular Biophysics, University of Maine, Orono, Maine 04469, USA.
Hess
Samuel T
ST
D000818:Animals; D002477:Cells / Q000737:chemistry* / Q000648:ultrastructure; D005347:Fibroblasts / Q000737:chemistry / Q000648:ultrastructure; D005454:Fluorescence Polarization / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000032:analysis; D006801:Humans; D008856:Microscopy, Fluorescence / Q000191:economics / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*
D049452:Green Fluorescent Proteins