Dataset to "Fluorescence Bar-coding and Flowmetry based on Dark State Transitions in Fluorescence Emitters"
type: Dataset
description: This folder contains all raw data underlying the results presented in a manuscript, submitted for publication to Journal of Physical Chemistry B, and entitled: Fluorescence Bar-coding and Flowmetry based on Dark State Transitions in Fluorescence Emitters Authored by: B. Demirbay‡, E. Sandberg‡, J. Piguet, J. Widengren* Royal Institute of Technology (KTH), Experimental Biomolecular Physics, Dept. Applied Physics, Albanova University Center 106 91 Stockholm, Sweden ‡Contributed equally. *Corresponding Author: Email: jwideng@kth.se, Phone: +46-8-7907813 The data files are grouped into the different techniques used to generate them, and refer to the figures/tables in the manuscript where the extracted results are presented. ABSTRACT Reversible dark state transitions in fluorophores represent a limiting factor in fluorescence-based ultrasensitive spectroscopy, is 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 establ…
conditions of access: https://publications.europa.eu/resource/authority/access-right/NON_PUBLIC
conditions of access: info:eu-repo/semantics/restrictedAccess
creative work status: completed
  type: DefinedTerm
  in defined term set: http://publications.europa.eu/resource/authority/dataset-status
  term code: COMPLETED
creator: Baris
  type: Person
  affiliation: Demirbay
  family name: Baris
creator: Elin
  type: Person
  affiliation: Sandberg
  family name: Elin
creator: Joachim
  type: Person
  affiliation: Piguet
  family name: Joachim
creator: Jerker
  type: Person
  affiliation: Widengren
  family name: Jerker
date modified: 2023-06-07
date published: 2023-06-05
is part of: https://zenodo.org/communities/kth/
keywords: Physical Sciences
  type: DefinedTerm
  term code: 103
keywords: photo-isomerization, transient state monitoring, fluorescence correlation spectroscopy, multiplexing, cellular imaging, microfluidics
publisher: Royal Institute of Technology
version: 0

PropertyValue
property ID: DOI
value: 10.5281/zenodo.8006451

PropertyValue
property ID: URL
value: https://zenodo.org/records/8006451

PropertyValue
property ID: oai
value: oai:zenodo.org:8006451

Royal Institute of Technology
type: Organization