Editing Optical microscope (section)

====Localization microscopy SPDMphymod====
[[File:3D Dual Color Super Resolution Microscopy Cremer 2010.png|thumb|600px|alt=3D Dual Color Super Resolution Microscopy Cremer from 2010|3D dual color super resolution microscopy with Her2 and Her3 in breast cells, standard dyes: Alexa 488, Alexa 568 LIMON]]
SPDM (spectral precision distance microscopy), the basic localization microscopy technology is a light optical process of [[fluorescence microscopy]] which allows position, distance and angle measurements on "optically isolated" particles (e.g. molecules) well below the theoretical [[limit of resolution]] for light microscopy. "Optically isolated" means that at a given point in time, only a single particle/molecule within a region of a size determined by conventional optical resolution (typically approx. 200–250&nbsp;nm [[diameter]]) is being registered. This is possible when [[molecules]] within such a region all carry different spectral markers (e.g. different colors or other usable differences in the [[light emission]] of different particles).<ref>{{cite journal |doi=10.1007/s00340-008-3152-x|title=SPDM: light microscopy with single-molecule resolution at the nanoscale|year=2008|journal=Applied Physics B|volume=93|issue=1|pages=1–12|last1=Lemmer|first1=P.|last2=Gunkel|first2=M.|last3=Baddeley|first3=D.|last4=Kaufmann|first4=R.|last5=Urich|first5=A.|last6=Weiland|first6=Y.|last7=Reymann|first7=J.|last8=Müller|first8=P.|last9=Hausmann|first9=M.|last10=Cremer|first10=C.|bibcode=2008ApPhB..93....1L|s2cid=13805053 }}</ref><ref>{{cite book|doi=10.1117/12.260797|chapter=Comparative study of three-dimensional localization accuracy in conventional, confocal laser scanning and axial tomographic fluorescence light microscopy|year=1996|last1=Bradl|first1=Joachim|editor5-first=Pierre M|editor5-last=Viallet|editor4-first=Katarina|editor4-last=Svanberg|editor3-first=Herbert|editor3-last=Schneckenburger|editor2-first=Warren S|editor2-last=Grundfest|editor1-first=Irving J|editor1-last=Bigio|title=Optical Biopsies and Microscopic Techniques|volume=2926|pages=201–206|series=Optical Biopsies and Microscopic Techniques|s2cid=55468495 }}</ref><ref>{{cite journal|author1=Heintzmann, R.|author2=Münch, H.|author3=Cremer, C.|year=1997|title=High-precision measurements in epifluorescent microscopy – simulation and experiment|journal=Cell Vision|volume=4|pages=252–253|url=http://www.kip.uni-heidelberg.de/AG_Cremer/sites/default/files/Bilder/pdf_1997/CellVisionVol4No2Heintzmann.pdf|url-status=live|archive-url=https://web.archive.org/web/20160216030456/http://www.kip.uni-heidelberg.de/AG_Cremer/sites/default/files/Bilder/pdf_1997/CellVisionVol4No2Heintzmann.pdf|archive-date=16 February 2016}}</ref><ref>Cremer, Christoph; Hausmann, Michael; Bradl, Joachim and Rinke, Bernd "Method and devices for measuring distances between object structures", {{US patent|6424421}} priority date 23 December 1996</ref>

Many standard fluorescent dyes like [[Green fluorescent protein|GFP]], Alexa dyes, Atto dyes, Cy2/Cy3 and fluorescein molecules can be used for localization microscopy, provided certain photo-physical conditions are present. Using this so-called SPDMphymod (physically modifiable fluorophores) technology a single laser wavelength of suitable intensity is sufficient for nanoimaging.<ref>{{cite journal|author=Manuel Gunkel|pmid=19548231|year=2009|title=Dual color localization microscopy of cellular nanostructures|volume=4|issue=6|pages=927–38|doi=10.1002/biot.200900005|journal=Biotechnology Journal|s2cid=18162278 |display-authors=etal|url=https://hal.archives-ouvertes.fr/hal-00494027/file/PEER_stage2_10.1002%252Fbiot.200900005.pdf |archive-url=https://web.archive.org/web/20190503232308/https://hal.archives-ouvertes.fr/hal-00494027/file/PEER_stage2_10.1002%252Fbiot.200900005.pdf |archive-date=2019-05-03 |url-status=live}}</ref>