Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/114092
Citations
Scopus Web of Science® Altmetric
?
?
Type: Journal article
Title: Super-multiplexed fluorescence microscopy via photostability contrast
Author: Orth, A.
Ghosh, R.
Wilson, E.
Doughney, T.
Brown, H.
Reineck, P.
Thompson, J.
Gibson, B.
Citation: Biomedical Optics Express, 2018; 9(7):2943-2954
Publisher: OSA Publishing
Issue Date: 2018
ISSN: 2156-7085
2156-7085
Statement of
Responsibility: 
Antony Orth, Richik N. Ghosh, Emma R. Wilson, Timothy Doughney, Hannah Brown, Philipp Reineck, Jeremy G. Thompson and Brant C. Gibson
Abstract: Fluorescence microscopy is widely used to observe and quantify the inner workings of the cell. Traditionally, multiple types of cellular structures or biomolecules are visualized simultaneously in a sample by using spectrally distinct fluorescent labels. The wide emission spectra of most fluorophores limits spectral multiplexing to four or five labels in a standard fluorescence microscope. Further multiplexing requires another dimension of contrast. Here, we show that photostability differences can be used to distinguish between fluorescent labels. By combining photobleaching characteristics with a novel unmixing algorithm, we resolve up to three fluorescent labels in a single spectral channel and unmix fluorescent labels with nearly identical emission spectra. We apply our technique to organic dyes, autofluorescent biomolecules and fluorescent proteins. Our approach has the potential to triple the multiplexing capabilities of any digital widefield or confocal fluorescence microscope with no additional hardware, making it readily accessible to a wide range of researchers.
Keywords: (110.4234) Multispectral and hyperspectral imaging; (170.2520) Fluorescence microscopy
Rights: © 2018 Optical Society of America under the terms of the OSA Open Access Publishing Agreement
RMID: 0030094632
DOI: 10.1364/BOE.9.002943
Grant ID: http://purl.org/au-research/grants/arc/CE140100003
http://purl.org/au-research/grants/arc/FT110100225
Appears in Collections:Medicine publications

Files in This Item:
There are no files associated with this item.


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.