Wide-field time-gated photoluminescence microscopy for fast ultrahigh-sensitivity imaging of photoluminescent probes

Razali, WAW; Sreenivasan, VKA; Bradac, C; Connor, M; Goldys, EM; Zvyagin, AV

Wide-field time-gated photoluminescence microscopy for fast ultrahigh-sensitivity imaging of photoluminescent probes

Razali, WAW Sreenivasan, VKA Bradac, C

Connor, M Goldys, EM Zvyagin, AV

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Publication Type:: Journal Article
Citation:: Journal of biophotonics, 2016, 9 (8), pp. 848 - 858
Issue Date:: 2016-08-01

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Field	Value	Language
dc.contributor.author	Razali, WAW	en_US
dc.contributor.author	Sreenivasan, VKA	en_US
dc.contributor.author	Bradac, C https://orcid.org/0000-0002-6673-7238	en_US
dc.contributor.author	Connor, M	en_US
dc.contributor.author	Goldys, EM	en_US
dc.contributor.author	Zvyagin, AV	en_US
dc.date.issued	2016-08-01	en_US
dc.identifier.citation	Journal of biophotonics, 2016, 9 (8), pp. 848 - 858	en_US
dc.identifier.uri	http://hdl.handle.net/10453/122841
dc.description.abstract	© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Fluorescence microscopy is a fundamental technique for the life sciences, where biocompatible and photostable photoluminescence probes in combination with fast and sensitive imaging systems are continually transforming this field. A wide-field time-gated photoluminescence microscopy system customised for ultrasensitive imaging of unique nanoruby probes with long photoluminescence lifetime is described. The detection sensitivity derived from the long photoluminescence lifetime of the nanoruby makes it possible to discriminate signals from unwanted autofluorescence background and laser backscatter by employing a time-gated image acquisition mode. This mode enabled several-fold improvement of the photoluminescence imaging contrast of discrete nanorubies dispersed on a coverslip. It enabled recovery of the photoluminescence signal emanating from discrete nanorubies when covered by a layer of an organic fluorescent dye, which were otherwise invisible without the use of spectral filtering approaches. Time-gated imaging also facilitated high sensitivity detection of nanorubies in a biological environment of cultured cells. Finally, we monitor the binding kinetics of nanorubies to a functionalised substrate, which exemplified a real-time assay in biological fluids. 3D-pseudo colour images of nanorubies immersed in a highly fluorescent dye solution. Nanoruby photoluminescence is subdued by that of the dye in continuous excitation/imaging (left), however it can be recovered by time-gated imaging (right). At the bottom is schematic diagram of nanoruby assay in a biological fluid.	en_US
dc.relation.ispartof	Journal of biophotonics	en_US
dc.relation.isbasedon	10.1002/jbio.201600050	en_US
dc.subject.classification	Optoelectronics & Photonics	en_US
dc.subject.mesh	Cell Line, Tumor	en_US
dc.subject.mesh	Animals	en_US
dc.subject.mesh	Mice	en_US
dc.subject.mesh	Fluorescent Dyes	en_US
dc.subject.mesh	Microscopy, Fluorescence	en_US
dc.subject.mesh	Luminescent Measurements	en_US
dc.subject.mesh	Lasers	en_US
dc.subject.mesh	Light	en_US
dc.title	Wide-field time-gated photoluminescence microscopy for fast ultrahigh-sensitivity imaging of photoluminescent probes	en_US
dc.type	Journal Article
utslib.citation.volume	8	en_US
utslib.citation.volume	9	en_US
utslib.for	0205 Optical Physics	en_US
utslib.for	0304 Medicinal and Biomolecular Chemistry	en_US
utslib.for	1004 Medical Biotechnology	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Mathematical and Physical Sciences
pubs.organisational-group	/University of Technology Sydney/Strength - IBMD - Initiative for Biomedical Devices
utslib.copyright.status	closed_access
pubs.issue	8	en_US
pubs.publication-status	Published	en_US
pubs.volume	9	en_US

Abstract:

© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Fluorescence microscopy is a fundamental technique for the life sciences, where biocompatible and photostable photoluminescence probes in combination with fast and sensitive imaging systems are continually transforming this field. A wide-field time-gated photoluminescence microscopy system customised for ultrasensitive imaging of unique nanoruby probes with long photoluminescence lifetime is described. The detection sensitivity derived from the long photoluminescence lifetime of the nanoruby makes it possible to discriminate signals from unwanted autofluorescence background and laser backscatter by employing a time-gated image acquisition mode. This mode enabled several-fold improvement of the photoluminescence imaging contrast of discrete nanorubies dispersed on a coverslip. It enabled recovery of the photoluminescence signal emanating from discrete nanorubies when covered by a layer of an organic fluorescent dye, which were otherwise invisible without the use of spectral filtering approaches. Time-gated imaging also facilitated high sensitivity detection of nanorubies in a biological environment of cultured cells. Finally, we monitor the binding kinetics of nanorubies to a functionalised substrate, which exemplified a real-time assay in biological fluids. 3D-pseudo colour images of nanorubies immersed in a highly fluorescent dye solution. Nanoruby photoluminescence is subdued by that of the dye in continuous excitation/imaging (left), however it can be recovered by time-gated imaging (right). At the bottom is schematic diagram of nanoruby assay in a biological fluid.

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http://hdl.handle.net/10453/122841