Linked Life Data

16314572

Source:http://linkedlifedata.com/resource/pubmed/id/16314572

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
49
pubmed:dateCreated
2005-12-7
pubmed:abstractText
Fluorescence microscopy is indispensable in many areas of science, but until recently, diffraction has limited the resolution of its lens-based variant. The diffraction barrier has been broken by a saturated depletion of the marker's fluorescent state by stimulated emission, but this approach requires picosecond laser pulses of GW/cm2 intensity. Here, we demonstrate the surpassing of the diffraction barrier in fluorescence microscopy with illumination intensities that are eight orders of magnitude smaller. The subdiffraction resolution results from reversible photoswitching of a marker protein between a fluorescence-activated and a nonactivated state, whereby one of the transitions is accomplished by means of a spatial intensity distribution featuring a zero. After characterizing the switching kinetics of the used marker protein asFP595, we demonstrate the current capability of this RESOLFT (reversible saturable optical fluorescence transitions) type of concept to resolve 50-100 nm in the focal plane. The observed resolution is limited only by the photokinetics of the protein and the perfection of the zero. Our results underscore the potential to finally achieve molecular resolution in fluorescence microscopy by technical optimization.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-10810003, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-10840057, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-10852900, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-10899992, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-12496281, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-12524551, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-12962391, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-14595350, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-14595362, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-15550670, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-15823037, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-15884061, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-15904066, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-15904340, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-15972810, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-15983383, http://linkedlifedata.com/resource/pubmed/commentcorrection/16314572-16135569
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0027-8424
pubmed:author
pubmed:issnType
Print
pubmed:day
6
pubmed:volume
102
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
17565-9
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
2005
pubmed:articleTitle
Breaking the diffraction barrier in fluorescence microscopy at low light intensities by using reversibly photoswitchable proteins.
pubmed:affiliation
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, D-37077 Göttingen, Germany.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't