Fluorescence correlation spectroscopy with a total internal reflection fluorescence STED microscope (TIRF-STED-FCS)
We characterize a novel fluorescence microscope which combines the high spatial discrimination of a total internal reflection epi-fluorescence (epi-TIRF) microscope with that of stimulated emission depletion (STED) nanoscopy. This combination of high axial confinement and dynamic-active lateral spat...
Saved in:
| Main Authors: | , |
|---|---|
| Format: | Article (Journal) |
| Language: | English |
| Published: |
17 February 2012
|
| In: |
Optics express
Year: 2012, Volume: 20, Issue: 5, Pages: 5243-5263 |
| ISSN: | 1094-4087 |
| DOI: | 10.1364/OE.20.005243 |
| Online Access: | Verlag, Volltext: http://dx.doi.org/10.1364/OE.20.005243 Verlag, Volltext: https://www.osapublishing.org/oe/abstract.cfm?uri=oe-20-5-5243 
|
| Author Notes: | Marcel Leutenegger, Christian Ringemann, Theo Lasser, Stefan W. Hell, and Christian Eggeling |
MARC
| LEADER | 00000caa a2200000 c 4500 | ||
|---|---|---|---|
| 001 | 1582393397 | ||
| 003 | DE-627 | ||
| 005 | 20220815042934.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 181029s2012 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1364/OE.20.005243 |2 doi | |
| 035 | |a (DE-627)1582393397 | ||
| 035 | |a (DE-576)512393397 | ||
| 035 | |a (DE-599)BSZ512393397 | ||
| 035 | |a (OCoLC)1341022010 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rda | ||
| 041 | |a eng | ||
| 084 | |a 30 |2 sdnb | ||
| 100 | 1 | |a Leutenegger, Marcel |e VerfasserIn |0 (DE-588)1170115756 |0 (DE-627)1037103521 |0 (DE-576)512393362 |4 aut | |
| 245 | 1 | 0 | |a Fluorescence correlation spectroscopy with a total internal reflection fluorescence STED microscope (TIRF-STED-FCS) |c Marcel Leutenegger, Christian Ringemann, Theo Lasser, Stefan W. Hell, and Christian Eggeling |
| 264 | 1 | |c 17 February 2012 | |
| 300 | |a 21 | ||
| 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 29.10.2018 | ||
| 520 | |a We characterize a novel fluorescence microscope which combines the high spatial discrimination of a total internal reflection epi-fluorescence (epi-TIRF) microscope with that of stimulated emission depletion (STED) nanoscopy. This combination of high axial confinement and dynamic-active lateral spatial discrimination of the detected fluorescence emission promises imaging and spectroscopy of the structure and function of cell membranes at the macro-molecular scale. Following a full theoretical description of the sampling volume and the recording of images of fluorescent beads, we exemplify the performance and limitations of the TIRF-STED nanoscope with particular attention to the polarization state of the laser excitation light. We demonstrate fluorescence correlation spectroscopy (FCS) with the TIRF-STED nanoscope by observing the diffusion of dye molecules in aqueous solutions and of fluorescent lipid analogs in supported lipid bilayers in the presence of background signal. The nanoscope reduced the out-of-focus background signal. A lateral resolution down to 40-50 nm was attained which was ultimately limited by the low lateral signal-to-background ratio inherent to the confocal epi-TIRF scheme. Together with the estimated axial confinement of about 55 nm, our TIRF-STED nanoscope achieved an almost isotropic and less than 1 attoliter small all-optically induced measurement volume. | ||
| 650 | 4 | |a Circular polarization | |
| 650 | 4 | |a Confocal laser scanning microscopy | |
| 650 | 4 | |a Confocal microscopy | |
| 650 | 4 | |a Fluorescence microscopy | |
| 650 | 4 | |a Fluorescence spectroscopy | |
| 650 | 4 | |a High numerical aperture optics | |
| 700 | 1 | |a Hell, Stefan |d 1962- |e VerfasserIn |0 (DE-588)139815716 |0 (DE-627)613785681 |0 (DE-576)313195889 |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Optics express |d Washington, DC : Optica, 1997 |g 20(2012), 5, Seite 5243-5263 |h Online-Ressource |w (DE-627)302718370 |w (DE-600)1491859-6 |w (DE-576)079876579 |x 1094-4087 |7 nnas |a Fluorescence correlation spectroscopy with a total internal reflection fluorescence STED microscope (TIRF-STED-FCS) |
| 773 | 1 | 8 | |g volume:20 |g year:2012 |g number:5 |g pages:5243-5263 |g extent:21 |a Fluorescence correlation spectroscopy with a total internal reflection fluorescence STED microscope (TIRF-STED-FCS) |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1364/OE.20.005243 |x Verlag |x Resolving-System |3 Volltext |
| 856 | 4 | 0 | |u https://www.osapublishing.org/oe/abstract.cfm?uri=oe-20-5-5243 |x Verlag |3 Volltext |
| 951 | |a AR | ||
| 992 | |a 20181029 | ||
| 993 | |a Article | ||
| 994 | |a 2012 | ||
| 998 | |g 139815716 |a Hell, Stefan |m 139815716:Hell, Stefan |d 130000 |d 130001 |d 700000 |d 718000 |e 130000PH139815716 |e 130001PH139815716 |e 700000PH139815716 |e 718000PH139815716 |k 0/130000/ |k 1/130000/130001/ |k 0/700000/ |k 1/700000/718000/ |p 4 | ||
| 999 | |a KXP-PPN1582393397 |e 3030037452 | ||
| BIB | |a Y | ||
| SER | |a journal | ||
| JSO | |a {"name":{"displayForm":["Marcel Leutenegger, Christian Ringemann, Theo Lasser, Stefan W. Hell, and Christian Eggeling"]},"id":{"eki":["1582393397"],"doi":["10.1364/OE.20.005243"]},"origin":[{"dateIssuedKey":"2012","dateIssuedDisp":"17 February 2012"}],"relHost":[{"physDesc":[{"extent":"Online-Ressource"}],"name":{"displayForm":["Optica"]},"id":{"zdb":["1491859-6"],"eki":["302718370"],"issn":["1094-4087"]},"origin":[{"publisher":"Optica ; Optical Society of America, OSA","dateIssuedKey":"1997","dateIssuedDisp":"1997-","publisherPlace":"Washington, DC ; Washington, DC"}],"corporate":[{"role":"isb","roleDisplay":"Herausgebendes Organ","display":"Optica"},{"roleDisplay":"Herausgebendes Organ","display":"Optical Society of America","role":"isb"}],"language":["eng"],"recId":"302718370","disp":"Fluorescence correlation spectroscopy with a total internal reflection fluorescence STED microscope (TIRF-STED-FCS)Optics express","note":["Gesehen am 10.01.2022","Herausgebendes Organ bis 2021: Optical Society of America"],"type":{"media":"Online-Ressource","bibl":"periodical"},"part":{"pages":"5243-5263","issue":"5","year":"2012","extent":"21","volume":"20","text":"20(2012), 5, Seite 5243-5263"},"pubHistory":["1.1997 -"],"title":[{"title":"Optics express","subtitle":"the international electronic journal of optics","title_sort":"Optics express"}]}],"physDesc":[{"extent":"21 S."}],"person":[{"family":"Leutenegger","given":"Marcel","display":"Leutenegger, Marcel","roleDisplay":"VerfasserIn","role":"aut"},{"family":"Hell","given":"Stefan","display":"Hell, Stefan","roleDisplay":"VerfasserIn","role":"aut"}],"title":[{"title":"Fluorescence correlation spectroscopy with a total internal reflection fluorescence STED microscope (TIRF-STED-FCS)","title_sort":"Fluorescence correlation spectroscopy with a total internal reflection fluorescence STED microscope (TIRF-STED-FCS)"}],"recId":"1582393397","language":["eng"],"type":{"media":"Online-Ressource","bibl":"article-journal"},"note":["Gesehen am 29.10.2018"]} | ||
| SRT | |a LEUTENEGGEFLUORESCEN1720 | ||