Combined fluorescence fluctuation and spectrofluorometric measure-ments reveal a red-shifted, near-IR emissive photo-isomerized form of Cyanine 5
type: Dataset
distribution: Landing page for the dataset
description: This folder contains all raw data underlying the results presented in a manuscript, accepted for publication in International Journal of Molecular Sciences, and entitled: Combined fluorescence fluctuation and spectrofluorometric measurements reveal a red-shifted, near-IR emissive photo-isomerized form of Cyanine 5 Authored by: Elin Sandberg 1, Joachim Piguet 1, Haichun Liu 1 and Jerker Widengren 1,* 1 Experimental Biomolecular Physics, Department of Applied Physics, Royal Institute of Technology (KTH), Stockholm, Sweden * To whom correspondence should be addressed. Email: jwideng@kth.se. Tel: +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 Cyanine fluorophores are extensively used in fluorescence spectroscopy and imaging. Upon continuous exciation, especially at excitation conditions used in single-molecule and super-resolution experiments, photo-isomerized states of cyanines easily reach population probabilies of around 50%. Still, effects of photo-isomerization are largely ignored in such experiments. Here, we studied the photo-isomerization of the pentamethine Cyanine 5 (Cy5) by two similar, yet complementary means to follow fluorophore blinking dynamics: fluorescence correlation spectroscopy (FCS) and transient state (TRAST) excitation-modulation spectroscopy. Additionally, we combined TRAST and spectrofluorimetry (spectral-TRAST), whereby emission spectra of Cy5 were recorded upon different rectangular pulse-train excitations. We also developed a framework for analyzing transitions between multiple emissive states in FCS and TRAST experiments, how the brightness of the different states is weighted, and what initial conditions that apply. Our FCS, TRAST and spectral-TRAST experiments showed significant differences in dark state relaxation amplitudes for different spectral detection ranges, which we attribute to an ad…
modified: 2023-01-17T00:00:00Z
issued: 2022-12-06T00:00:00Z
publisher: Royal Institute of Technology
access rights: PUBLIC
qualified attribution: Attribution
  had role: principalInvestigator
  agent: Elin Sandberg
    type: Agent
    account: 0000-0003-3252-694X
    type: PrivateIndividual(s)
qualified attribution: Attribution
  had role: principalInvestigator
  agent: Joachim Piguet
    type: Agent
    account: https://orcid.org/0000-0002-4762-4887
    type: PrivateIndividual(s)
qualified attribution: Attribution
  had role: principalInvestigator
  agent: Haichun Liu
    type: Agent
    account: https://orcid.org/0000-0002-1780-7746
    type: PrivateIndividual(s)
qualified attribution: Attribution
  had role: principalInvestigator
  agent: Jerker Widengren
    type: Agent
    account: https://orcid.org/0000-0003-3200-0374
    type: PrivateIndividual(s)
keyword: Cyanine
keyword: isomerization
keyword: FCS
keyword: TRAST
keyword: Cy5
keyword: single molecule
keyword: emission red-shift
keyword: near-IR
keyword: photo-dynamics
keyword: excitation modulation
language: ENG

Landing page for the dataset
type: Distribution
title: Landningssida för datasetet @sv
access URL: Combined fluorescence fluctuation and spectrofluorometric measure-ments reveal a red-shifted, near-IR emissive photo-isomerized form of Cyanine 5
availability: stable

Royal Institute of Technology
type: Agent
name: Kungliga Tekniska högskolan @sv
type: Academia-ScientificOrganisation
source: 026vcq606