Photoswitches: Key molecules for subdiffraction-resolution fluorescence imaging and molecular quantification Export

@article{citeulike:4271078, abstract = {Optical microscopes, often referred to as lsquolight microscopesrsquo, use visible light and a system of lenses to provide us with magnified images of small samples. Combined with highly sensitive fluorescence detection techniques and efficient fluorescent probes they allow the non-invasive 3D study of subcellular structures even in living cells or tissue. However, optical microscopes are subject to the diffraction barrier of light which imposes an optical resolution limit of approximately 200 nm in the imaging plane. In the recent past new techniques emerged that break the diffraction barrier and enable structural investigations with so far unmatched resolution. They are all based on the selective switching of fluorophores between a fluorescent and a nonfluorescent state and are therefore generalized under the denotation ldquoPhotoswitching Microscopyrdquo. Here we review recent progress in subdiffraction-resolution fluorescence imaging microscopy using various photoswitchable fluorophores and strategies. Special emphasis will be placed on the design and development of photoswitches and the requirements photoswitches have to fulfill for successful use in photoswitching microscopy. Moreover, we demonstrate how photoswitches can be used advantageously for molecular quantification, i.e. the determination of densities and absolute numbers of proteins located in specific subcellular compartments and discuss concepts how standard organic fluorophores can be used successfully for photoswitching microscopy.}, address = {Applied Laser Physics and Laser Spectroscopy and Bielefeld Institute for Biophysics and Nanosciene, Bielefeld University, Universit\"{a}tsstr. 25, 33615 Bielefeld, Germany; Department of Chemistry, Katholieke Universiteit Leuven, Celestijnenlaan 200F, 3001 Heverlee, Belgium}, author = {Heilemann, M. and Dedecker, P. and Hofkens, J. and Sauer, M.}, citeulike-article-id = {4271078}, citeulike-linkout-0 = {http://dx.doi.org/10.1002/lpor.200810043}, citeulike-linkout-1 = {http://www3.interscience.wiley.com/cgi-bin/abstract/121633922/ABSTRACT}, doi = {10.1002/lpor.200810043}, issn = {1863-8899}, journal = {Laser \& Photonics Review}, keywords = {fluorophores, photoswitchable, review, super-resolution}, number = {1-2}, pages = {180--202}, posted-at = {2009-04-03 20:58:51}, priority = {2}, title = {Photoswitches: Key molecules for subdiffraction-resolution fluorescence imaging and molecular quantification}, url = {http://dx.doi.org/10.1002/lpor.200810043}, volume = {3}, year = {2009} }

TY - JOUR ID - citeulike:4271078 L3 - citeulike-article-id:4271078 N2 - Optical microscopes, often referred to as lsquolight microscopesrsquo, use visible light and a system of lenses to provide us with magnified images of small samples. Combined with highly sensitive fluorescence detection techniques and efficient fluorescent probes they allow the non-invasive 3D study of subcellular structures even in living cells or tissue. However, optical microscopes are subject to the diffraction barrier of light which imposes an optical resolution limit of approximately 200 nm in the imaging plane. In the recent past new techniques emerged that break the diffraction barrier and enable structural investigations with so far unmatched resolution. They are all based on the selective switching of fluorophores between a fluorescent and a nonfluorescent state and are therefore generalized under the denotation ldquoPhotoswitching Microscopyrdquo. Here we review recent progress in subdiffraction-resolution fluorescence imaging microscopy using various photoswitchable fluorophores and strategies. Special emphasis will be placed on the design and development of photoswitches and the requirements photoswitches have to fulfill for successful use in photoswitching microscopy. Moreover, we demonstrate how photoswitches can be used advantageously for molecular quantification, i.e. the determination of densities and absolute numbers of proteins located in specific subcellular compartments and discuss concepts how standard organic fluorophores can be used successfully for photoswitching microscopy. IS - 1-2 JF - Laser & Photonics Review SN - 1863-8899 AD - Applied Laser Physics and Laser Spectroscopy and Bielefeld Institute for Biophysics and Nanosciene, Bielefeld University, Universitätsstr. 25, 33615 Bielefeld, Germany; Department of Chemistry, Katholieke Universiteit Leuven, Celestijnenlaan 200F, 3001 Heverlee, Belgium EP - 202 TI - Photoswitches: Key molecules for subdiffraction-resolution fluorescence imaging and molecular quantification VL - 3 SP - 180 KW - fluorophores KW - photoswitchable KW - review KW - super-resolution AU - Heilemann, M AU - Dedecker, P AU - Hofkens, J AU - Sauer, M PY - 2009/// UR - http://dx.doi.org/10.1002/lpor.200810043 ER -