Experimental approach to extend the range for counting fluorescent molecules based on photon-antibunching
In single-molecule fluorescence spectroscopy photon-antibunching is frequently used to prove the occurrence of single fluorophores. Furthermore, the relative frequency of coincident photon pairs was also used to determine the number of fluorophores in the diffraction limited observation volume of a...
Gespeichert in:
| Hauptverfasser: | , , , |
|---|---|
| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
06 Jul 2010
|
| In: |
Physical chemistry, chemical physics
Year: 2010, Jahrgang: 12, Heft: 35, Pages: 10295-10300 |
| ISSN: | 1463-9084 |
| DOI: | 10.1039/C0CP00363H |
| Online-Zugang: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1039/C0CP00363H Verlag, lizenzpflichtig, Volltext: https://pubs.rsc.org/en/content/articlelanding/2010/cp/c0cp00363h 
|
| Verfasserangaben: | Haisen Ta, Alexander Kiel, Michael Wahl and Dirk-Peter Herten |
MARC
| LEADER | 00000caa a2200000 c 4500 | ||
|---|---|---|---|
| 001 | 1859416055 | ||
| 003 | DE-627 | ||
| 005 | 20240311124138.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 230913s2010 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1039/C0CP00363H |2 doi | |
| 035 | |a (DE-627)1859416055 | ||
| 035 | |a (DE-599)KXP1859416055 | ||
| 035 | |a (OCoLC)1425873777 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rda | ||
| 041 | |a eng | ||
| 084 | |a 30 |2 sdnb | ||
| 100 | 1 | |a Ta, Haisen |e VerfasserIn |0 (DE-588)141019859 |0 (DE-627)703859102 |0 (DE-576)322225817 |4 aut | |
| 245 | 1 | 0 | |a Experimental approach to extend the range for counting fluorescent molecules based on photon-antibunching |c Haisen Ta, Alexander Kiel, Michael Wahl and Dirk-Peter Herten |
| 264 | 1 | |c 06 Jul 2010 | |
| 300 | |a 6 | ||
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Gesehen am 13.09.2023 | ||
| 520 | |a In single-molecule fluorescence spectroscopy photon-antibunching is frequently used to prove the occurrence of single fluorophores. Furthermore, the relative frequency of coincident photon pairs was also used to determine the number of fluorophores in the diffraction limited observation volume of a confocal microscope. However, the ability to count fluorophores is so far limited to ∼3 molecules due to saturation of the calibration curve with increasing number of fluorophores. Recently, we introduced a novel theoretical framework for counting the number of emitting molecules by analyzing photon-distributions acquired with a confocal microscope using four single-photon detectors. Here, we present the experimental realization of the proposed scheme in a confocal setup using novel multi-channel photon-counting electronics and DNA constructs that were labelled with five fluorophores. Our experimental results give a clear correlation between the number of estimated fluorophores and the number of bleaching steps for DNA probes conjugated with five ATTO647N labels with an error of ∼20%. Moreover, we could acquire experimental data for up to 15 fluorophores indicating the simultaneous occurrence of three DNA probes. Our experiments put into perspective that the analysis of photon-distributions acquired with four detection channels is suited to count the number of fluorescently labelled molecules in larger aggregates or clusters with potential for applications in molecular and cell biology and for time-resolved analysis of multi-chromophoric compounds in material sciences. | ||
| 700 | 1 | |a Kiel, Alexander |e VerfasserIn |0 (DE-588)135883172 |0 (DE-627)572477236 |0 (DE-576)300698828 |4 aut | |
| 700 | 1 | |a Wahl, Michael |e VerfasserIn |4 aut | |
| 700 | 1 | |a Herten, Dirk-Peter |e VerfasserIn |0 (DE-588)1020454768 |0 (DE-627)691223882 |0 (DE-576)360394051 |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Physical chemistry, chemical physics |d Cambridge : RSC Publ., 1999 |g 12(2010), 35, Seite 10295-10300 |h Online-Ressource |w (DE-627)270125175 |w (DE-600)1476244-4 |w (DE-576)080160328 |x 1463-9084 |7 nnas |a Experimental approach to extend the range for counting fluorescent molecules based on photon-antibunching |
| 773 | 1 | 8 | |g volume:12 |g year:2010 |g number:35 |g pages:10295-10300 |g extent:6 |a Experimental approach to extend the range for counting fluorescent molecules based on photon-antibunching |
| 776 | 0 | 8 | |i Erscheint auch als |n Druck-Ausgabe |a Ta, Haisen |t Experimental approach to extend the range for counting fluorescent molecules based on photon-antibunching |d 2010 |w (DE-627)1629524085 |w (DE-576)36067822X |
| 856 | 4 | 0 | |u https://doi.org/10.1039/C0CP00363H |x Verlag |x Resolving-System |z lizenzpflichtig |3 Volltext |
| 856 | 4 | 0 | |u https://pubs.rsc.org/en/content/articlelanding/2010/cp/c0cp00363h |x Verlag |z lizenzpflichtig |3 Volltext |
| 951 | |a AR | ||
| 992 | |a 20230913 | ||
| 993 | |a Article | ||
| 994 | |a 2010 | ||
| 998 | |g 1020454768 |a Herten, Dirk-Peter |m 1020454768:Herten, Dirk-Peter |d 120000 |d 700000 |d 718000 |e 120000PH1020454768 |e 700000PH1020454768 |e 718000PH1020454768 |k 0/120000/ |k 0/700000/ |k 1/700000/718000/ |p 4 |y j | ||
| 998 | |g 135883172 |a Kiel, Alexander |m 135883172:Kiel, Alexander |d 700000 |d 718000 |e 700000PK135883172 |e 718000PK135883172 |k 0/700000/ |k 1/700000/718000/ |p 2 | ||
| 998 | |g 141019859 |a Ta, Haisen |m 141019859:Ta, Haisen |d 120000 |e 120000PT141019859 |k 0/120000/ |p 1 |x j | ||
| 999 | |a KXP-PPN1859416055 |e 4375633107 | ||
| BIB | |a Y | ||
| SER | |a journal | ||
| JSO | |a {"person":[{"family":"Ta","display":"Ta, Haisen","given":"Haisen","role":"aut"},{"role":"aut","display":"Kiel, Alexander","given":"Alexander","family":"Kiel"},{"role":"aut","given":"Michael","display":"Wahl, Michael","family":"Wahl"},{"role":"aut","family":"Herten","display":"Herten, Dirk-Peter","given":"Dirk-Peter"}],"name":{"displayForm":["Haisen Ta, Alexander Kiel, Michael Wahl and Dirk-Peter Herten"]},"origin":[{"dateIssuedDisp":"06 Jul 2010","dateIssuedKey":"2010"}],"recId":"1859416055","title":[{"title_sort":"Experimental approach to extend the range for counting fluorescent molecules based on photon-antibunching","title":"Experimental approach to extend the range for counting fluorescent molecules based on photon-antibunching"}],"language":["eng"],"physDesc":[{"extent":"6 S."}],"id":{"doi":["10.1039/C0CP00363H"],"eki":["1859416055"]},"type":{"bibl":"article-journal","media":"Online-Ressource"},"note":["Gesehen am 13.09.2023"],"relHost":[{"type":{"bibl":"periodical","media":"Online-Ressource"},"pubHistory":["1.1999 -"],"titleAlt":[{"title":"PCCP"}],"language":["eng"],"recId":"270125175","disp":"Experimental approach to extend the range for counting fluorescent molecules based on photon-antibunchingPhysical chemistry, chemical physics","note":["Gesehen am 20.02.24"],"part":{"year":"2010","pages":"10295-10300","volume":"12","text":"12(2010), 35, Seite 10295-10300","extent":"6","issue":"35"},"id":{"zdb":["1476244-4"],"issn":["1463-9084"],"eki":["270125175"]},"physDesc":[{"extent":"Online-Ressource"}],"title":[{"title":"Physical chemistry, chemical physics","title_sort":"Physical chemistry, chemical physics","subtitle":"PCCP ; a journal of European chemical societies"}],"corporate":[{"display":"Royal Society of Chemistry","role":"isb"}],"origin":[{"publisherPlace":"Cambridge","dateIssuedKey":"1999","dateIssuedDisp":"1999-","publisher":"RSC Publ."}],"name":{"displayForm":["RSC, Royal Society of Chemistry"]}}]} | ||
| SRT | |a TAHAISENKIEXPERIMENT0620 | ||