Hyperspectral imaging for simultaneous measurements of two FRET biosensors in pancreatic β-cells

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dc.citation.articleNumbere0188789
dc.citation.issueNumber12
dc.citation.journalTitlePLoS ONE
dc.citation.volumeNumber12
dc.contributor.authorElliott, Amicia D.
dc.contributor.authorBedard, Noah
dc.contributor.authorUstione, Alessandro
dc.contributor.authorBaird, Michelle A.
dc.contributor.authorDavidson, Michael W.
dc.contributor.authorTkaczyk, Tomasz
dc.contributor.authorPiston, David W.
dc.date.accessioned2017-12-18T18:19:42Z
dc.date.available2017-12-18T18:19:42Z
dc.date.issued2017
dc.description.abstractFluorescent protein (FP) biosensors based on Förster resonance energy transfer (FRET) are commonly used to study molecular processes in living cells. There are FP-FRET biosensors for many cellular molecules, but it remains difficult to perform simultaneous measurements of multiple biosensors. The overlapping emission spectra of the commonly used FPs, including CFP/YFP and GFP/RFP make dual FRET measurements challenging. In addition, a snapshot imaging modality is required for simultaneous imaging. The Image Mapping Spectrometer (IMS) is a snapshot hyperspectral imaging system that collects high resolution spectral data and can be used to overcome these challenges. We have previously demonstrated the IMS’s capabilities for simultaneously imaging GFP and CFP/YFP-based biosensors in pancreatic β-cells. Here, we demonstrate a further capability of the IMS to image simultaneously two FRET biosensors with a single excitation band, one for cAMP and the other for Caspase-3. We use these measurements to measure simultaneously cAMP signaling and Caspase-3 activation in pancreatic β-cells during oxidative stress and hyperglycemia, which are essential components in the pathology of diabetes.
dc.identifier.citationElliott, Amicia D., Bedard, Noah, Ustione, Alessandro, et al.. "Hyperspectral imaging for simultaneous measurements of two FRET biosensors in pancreatic β-cells." <i>PLoS ONE,</i> 12, no. 12 (2017) Public Library of Science: https://doi.org/10.1371/journal.pone.0188789.
dc.identifier.digitaljournal-pone-0188789
dc.identifier.doihttps://doi.org/10.1371/journal.pone.0188789
dc.identifier.urihttps://hdl.handle.net/1911/98889
dc.language.isoeng
dc.publisherPublic Library of Science
dc.rightsThis is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.
dc.rights.urihttps://creativecommons.org/publicdomain/zero/1.0/
dc.titleHyperspectral imaging for simultaneous measurements of two FRET biosensors in pancreatic β-cells
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpublisher version
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