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| Fältnamn | Indikatorer | Metadata |
|---|---|---|
| 000 | 03956naa a2200361 4500 | |
| 001 | oai:DiVA.org:kth-342739 | |
| 003 | SwePub | |
| 008 | 240206s2024 | |||||||||||000 ||eng| | |
| 024 | 7 | a https://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-3427392 URI |
| 024 | 7 | a https://doi.org/10.1021/acs.jpcb.3c069052 DOI |
| 040 | a (SwePub)kth | |
| 041 | a engb eng | |
| 042 | 9 SwePub | |
| 072 | 7 | a ref2 swepub-contenttype |
| 072 | 7 | a art2 swepub-publicationtype |
| 100 | 1 | a Sandberg, Elinu KTH,Kvant- och biofotonik,Albanova University Center, 106 91 Stockholm, Sweden4 aut0 (Swepub:kth)u1643wp3 |
| 245 | 1 0 | a Fluorescence Bar-Coding and Flowmetry Based on Dark State Transitions in Fluorescence Emitters |
| 264 | 1 | b American Chemical Society (ACS),c 2024 |
| 338 | a print2 rdacarrier | |
| 500 | a QC 20240206 | |
| 520 | a Reversible dark state transitions in fluorophores represent a limiting factor in fluorescence-based ultrasensitive spectroscopy, are a necessary basis for fluorescence-based super-resolution imaging, but may also offer additional, largely orthogonal fluorescence-based readout parameters. In this work, we analyzed the blinking kinetics of Cyanine5 (Cy5) as a bar-coding feature distinguishing Cy5 from rhodamine fluorophores having largely overlapping emission spectra. First, fluorescence correlation spectroscopy (FCS) solution measurements on mixtures of free fluorophores and fluorophore-labeled small unilamellar vesicles (SUVs) showed that Cy5 could be readily distinguished from the rhodamines by its reversible, largely excitation-driven trans-cis isomerization. This was next confirmed by transient state (TRAST) spectroscopy measurements, determining the fluorophore dark state kinetics in a more robust manner, from how the time-averaged fluorescence intensity varies upon modulation of the applied excitation light. TRAST was then combined with wide-field imaging of live cells, whereby Cy5 and rhodamine fluorophores could be distinguished on a whole cell level as well as in spatially resolved, multiplexed images of the cells. Finally, we established a microfluidic TRAST concept and showed how different mixtures of free Cy5 and rhodamine fluorophores and corresponding fluorophore-labeled SUVs could be distinguished on-the-fly when passing through a microfluidic channel. In contrast to FCS, TRAST does not rely on single-molecule detection conditions or a high time resolution and is thus broadly applicable to different biological samples. Therefore, we expect that the bar-coding concept presented in this work can offer an additional useful strategy for fluorescence-based multiplexing that can be implemented on a broad range of both stationary and moving samples. | |
| 650 | 7 | a NATURVETENSKAPx Biologix Biofysik0 (SwePub)106032 hsv//swe |
| 650 | 7 | a NATURAL SCIENCESx Biological Sciencesx Biophysics0 (SwePub)106032 hsv//eng |
| 700 | 1 | a Demirbay, Barisu KTH,Kvant- och biofotonik,Albanova University Center, 106 91 Stockholm, Sweden4 aut0 (Swepub:kth)u10bas52 |
| 700 | 1 | a Kulkarni, Abhilashu KTH,Tillämpad fysik,Albanova University Center, 106 91 Stockholm, Sweden4 aut0 (Swepub:kth)u1wrw20i |
| 700 | 1 | a Liu, Haichunu KTH,Kvant- och biofotonik,Albanova University Center, 106 91 Stockholm, Sweden4 aut0 (Swepub:kth)u1mn5cz0 |
| 700 | 1 | a Piguet, Joachim,d 1979-u KTH,Kvant- och biofotonik,Albanova University Center, 106 91 Stockholm, Sweden4 aut0 (Swepub:kth)u1s3pwbd |
| 700 | 1 | a Widengren, Jerkeru KTH,Kvant- och biofotonik,Albanova University Center, 106 91 Stockholm, Sweden4 aut0 (Swepub:kth)u1i3g09c |
| 710 | 2 | a KTHb Kvant- och biofotonik4 org |
| 773 | 0 | t Journal of Physical Chemistry Bd : American Chemical Society (ACS)g 128:1, s. 125-136q 128:1<125-136x 1520-6106x 1520-5207 |
| 856 | 4 | u https://doi.org/10.1021/acs.jpcb.3c06905y Fulltext |
| 856 | 4 8 | u https://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-342739 |
| 856 | 4 8 | u https://doi.org/10.1021/acs.jpcb.3c06905 |
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