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Snapshot multicolor fluorescence imaging using double multiplexing of excitation and emission on a single detector

Dorozynska, Karolina LU ; Ek, Simon LU ; Kornienko, Vassily LU ; Andersson, David LU ; Andersson, Alexandra LU ; Ehn, Andreas LU and Kristensson, Elias LU (2021) In Scientific Reports 11(1).
Abstract

Fluorescence-based multispectral imaging of rapidly moving or dynamic samples requires both fast two-dimensional data acquisition as well as sufficient spectral sensitivity for species separation. As the number of fluorophores in the experiment increases, meeting both these requirements becomes technically challenging. Although several solutions for fast imaging of multiple fluorophores exist, they all have one main restriction; they rely solely on spectrally resolving either the excitation- or the emission characteristics of the fluorophores. This inability directly limits how many fluorophores existing methods can simultaneously distinguish. Here we present a snapshot multispectral imaging approach that not only senses the excitation... (More)

Fluorescence-based multispectral imaging of rapidly moving or dynamic samples requires both fast two-dimensional data acquisition as well as sufficient spectral sensitivity for species separation. As the number of fluorophores in the experiment increases, meeting both these requirements becomes technically challenging. Although several solutions for fast imaging of multiple fluorophores exist, they all have one main restriction; they rely solely on spectrally resolving either the excitation- or the emission characteristics of the fluorophores. This inability directly limits how many fluorophores existing methods can simultaneously distinguish. Here we present a snapshot multispectral imaging approach that not only senses the excitation and emission characteristics of the probed fluorophores but also all cross term combinations of excitation and emission. To the best of the authors’ knowledge, this is the only snapshot multispectral imaging method that has this ability, allowing us to even sense and differentiate between light of equal wavelengths emitted from the same fluorescing species but where the signal components stem from different excitation sources. The current implementation of the technique allows us to simultaneously gather 24 different spectral images on a single detector, from which we demonstrate the ability to visualize and distinguish up to nine fluorophores within the visible wavelength range.

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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Scientific Reports
volume
11
issue
1
article number
20454
publisher
Nature Publishing Group
external identifiers
  • scopus:85117377050
  • pmid:34650144
ISSN
2045-2322
DOI
10.1038/s41598-021-99670-6
project
Advanced Laser Diagnostics for Discharge Plasma
language
English
LU publication?
yes
additional info
Publisher Copyright: © 2021, The Author(s).
id
a29856f8-26fa-4cba-8d0e-9654c422f743
date added to LUP
2021-11-19 13:07:08
date last changed
2024-06-15 20:43:44
@article{a29856f8-26fa-4cba-8d0e-9654c422f743,
  abstract     = {{<p>Fluorescence-based multispectral imaging of rapidly moving or dynamic samples requires both fast two-dimensional data acquisition as well as sufficient spectral sensitivity for species separation. As the number of fluorophores in the experiment increases, meeting both these requirements becomes technically challenging. Although several solutions for fast imaging of multiple fluorophores exist, they all have one main restriction; they rely solely on spectrally resolving either the excitation- or the emission characteristics of the fluorophores. This inability directly limits how many fluorophores existing methods can simultaneously distinguish. Here we present a snapshot multispectral imaging approach that not only senses the excitation and emission characteristics of the probed fluorophores but also all cross term combinations of excitation and emission. To the best of the authors’ knowledge, this is the only snapshot multispectral imaging method that has this ability, allowing us to even sense and differentiate between light of equal wavelengths emitted from the same fluorescing species but where the signal components stem from different excitation sources. The current implementation of the technique allows us to simultaneously gather 24 different spectral images on a single detector, from which we demonstrate the ability to visualize and distinguish up to nine fluorophores within the visible wavelength range.</p>}},
  author       = {{Dorozynska, Karolina and Ek, Simon and Kornienko, Vassily and Andersson, David and Andersson, Alexandra and Ehn, Andreas and Kristensson, Elias}},
  issn         = {{2045-2322}},
  language     = {{eng}},
  number       = {{1}},
  publisher    = {{Nature Publishing Group}},
  series       = {{Scientific Reports}},
  title        = {{Snapshot multicolor fluorescence imaging using double multiplexing of excitation and emission on a single detector}},
  url          = {{http://dx.doi.org/10.1038/s41598-021-99670-6}},
  doi          = {{10.1038/s41598-021-99670-6}},
  volume       = {{11}},
  year         = {{2021}},
}