Title: Time‐resolved identification of individual mononucleotide molecules in aqueous solution with pulsed semiconductor lasers
Abstract: BioimagingVolume 6, Issue 1 p. 14-24 Research ArticleFree Access Time-resolved identification of individual mononucleotide molecules in aqueous solution with pulsed semiconductor lasers Markus Sauer, Markus Sauer [email protected] Physikalisch-Chemisches Institut, Universität Heidelberg, Im Neuenheimer Feld 253, 69120 Heidelberg, GermanySearch for more papers by this authorJutta Arden-Jacob, Jutta Arden-Jacob Institut für Physikalische Chemie, Universität-Gesamthochschule Siegen, Adolf-Reichwein-Straße 3, 57068 Siegen, GermanySearch for more papers by this authorKarl H Drexhage, Karl H Drexhage Institut für Physikalische Chemie, Universität-Gesamthochschule Siegen, Adolf-Reichwein-Straße 3, 57068 Siegen, GermanySearch for more papers by this authorFlorian Göbel, Florian Göbel Physikalisch-Chemisches Institut, Universität Heidelberg, Im Neuenheimer Feld 253, 69120 Heidelberg, GermanySearch for more papers by this authorUlrike Lieberwirth, Ulrike Lieberwirth Physikalisch-Chemisches Institut, Universität Heidelberg, Im Neuenheimer Feld 253, 69120 Heidelberg, GermanySearch for more papers by this authorKlaus Mühlegger, Klaus Mühlegger Boehringer Mannheim GmbH, Werk Penzberg, Nonnenwald 2, 82377 Penzberg, GermanySearch for more papers by this authorRalph Müller, Ralph Müller Physikalisch-Chemisches Institut, Universität Heidelberg, Im Neuenheimer Feld 253, 69120 Heidelberg, GermanySearch for more papers by this authorJürgen Wolfrum, Jürgen Wolfrum Physikalisch-Chemisches Institut, Universität Heidelberg, Im Neuenheimer Feld 253, 69120 Heidelberg, GermanySearch for more papers by this authorChristoph Zander, Christoph Zander Physikalisch-Chemisches Institut, Universität Heidelberg, Im Neuenheimer Feld 253, 69120 Heidelberg, Germany Institut für Physikalische Chemie, Universität-Gesamthochschule Siegen, Adolf-Reichwein-Straße 3, 57068 Siegen, GermanySearch for more papers by this author Markus Sauer, Markus Sauer [email protected] Physikalisch-Chemisches Institut, Universität Heidelberg, Im Neuenheimer Feld 253, 69120 Heidelberg, GermanySearch for more papers by this authorJutta Arden-Jacob, Jutta Arden-Jacob Institut für Physikalische Chemie, Universität-Gesamthochschule Siegen, Adolf-Reichwein-Straße 3, 57068 Siegen, GermanySearch for more papers by this authorKarl H Drexhage, Karl H Drexhage Institut für Physikalische Chemie, Universität-Gesamthochschule Siegen, Adolf-Reichwein-Straße 3, 57068 Siegen, GermanySearch for more papers by this authorFlorian Göbel, Florian Göbel Physikalisch-Chemisches Institut, Universität Heidelberg, Im Neuenheimer Feld 253, 69120 Heidelberg, GermanySearch for more papers by this authorUlrike Lieberwirth, Ulrike Lieberwirth Physikalisch-Chemisches Institut, Universität Heidelberg, Im Neuenheimer Feld 253, 69120 Heidelberg, GermanySearch for more papers by this authorKlaus Mühlegger, Klaus Mühlegger Boehringer Mannheim GmbH, Werk Penzberg, Nonnenwald 2, 82377 Penzberg, GermanySearch for more papers by this authorRalph Müller, Ralph Müller Physikalisch-Chemisches Institut, Universität Heidelberg, Im Neuenheimer Feld 253, 69120 Heidelberg, GermanySearch for more papers by this authorJürgen Wolfrum, Jürgen Wolfrum Physikalisch-Chemisches Institut, Universität Heidelberg, Im Neuenheimer Feld 253, 69120 Heidelberg, GermanySearch for more papers by this authorChristoph Zander, Christoph Zander Physikalisch-Chemisches Institut, Universität Heidelberg, Im Neuenheimer Feld 253, 69120 Heidelberg, Germany Institut für Physikalische Chemie, Universität-Gesamthochschule Siegen, Adolf-Reichwein-Straße 3, 57068 Siegen, GermanySearch for more papers by this author First published: 11 May 2001 https://doi.org/10.1002/1361-6374(199803)6:1<14::AID-BIO3>3.0.CO;2-OCitations: 46AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Abstract We applied a short-pulse diode laser emitting at 640 nm with a repetition rate of 56 MHz in combination with a confocal microscope to study bursts of fluorescence photons from individual differently labeled mononucleotide molecules in water. Two newly synthesized dyes, an oxazine dye (MR121) and a rhodamine dye (JA53), and two commercially available dyes, a carbocyanine dye (Cy5) and a bora-diaza-indacene dye (Bodipy630/650), were used as fluorescent labels. The time-resolved fluorescence signals of individual mononucleotiode molecules in water were analyzed and identified by a maximum likelihood estimator (MLE). Taking only those single molecule transits which contain more than 30 collected photoelectrons, the two labeled mononucleotide molecules, Cy5-dCTP and Bodipy-dUTP, can be identified by time-resolved fluorescence spectroscopy with a probability of correct classification of greater than 99%. Our results show that at least three differently labeled mononucleotide molecules can be identified in a common aqueous solution. We obtain an overall classification probability of 90% for the time-resolved identification of Cy5-dCTP, MR121-dUTP and Bodipy-dUTP molecules via their characteristic fluorescence lifetimes of 1.05 ± 0.33 ns (Cy5-dCTP), 2.07 ± 0.59 ns (MR121-dUTP) and 3.88 ± 1.71 ns (Bodipy-dUTP). Citing Literature Volume6, Issue1Special Issue: Single Molecule Detection: Basics and Applications in Life SciencesMarch 1998Pages 14-24 RelatedInformation