Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates

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Title: Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates
Authors: Maioli, V
Chennell, G
Sparks, H
Lana, T
Kumar, S
Carling, D
Sardini, A
Dunsby, CW
Item Type: Journal Article
Abstract: Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals. As a proof-of-principle, the ssOPM system is also used to acquire a dose response curve with the concentration of glucose in the culture medium being varied across 42 wells of a 96-well plate with the whole acquisition taking 9 min. The 3-D image data enable the FRET ratio to be measured as a function of distance from the surface of the spheroid. Overall, the results demonstrate the capability of the OPM system to measure spatio-temporal changes in FRET ratio in 3-D in multicellular spheroids over time in a multi-well plate format.
Issue Date: 25-Nov-2016
Date of Acceptance: 1-Nov-2016
URI: http://hdl.handle.net/10044/1/42274
DOI: https://dx.doi.org/10.1038/srep37777
ISSN: 2045-2322
Publisher: Nature Publishing Group
Journal / Book Title: Scientific Reports
Volume: 6
Sponsor/Funder: The Royal Society
Engineering & Physical Science Research Council (EPSRC)
Biotechnology and Biological Sciences Research Council (BBSRC)
Engineering & Physical Science Research Council (E
Biotechnology and Biological Sciences Research Council (BBSRC)
Funder's Grant Number: N/A
EP/H03238X/1
BB/I023801/1
EP/K503733/1
BB/M006786/1
Keywords: Science & Technology
Multidisciplinary Sciences
Science & Technology - Other Topics
RESONANCE ENERGY-TRANSFER
OBLIQUE PLANE MICROSCOPY
IN-VITRO MODEL
CARDIAC MYOCYTES
DRUG DISCOVERY
LIVING CELLS
3D
CULTURES
PROTEIN
FRET
Publication Status: Published
Article Number: 37777
Appears in Collections:Physics
Photonics
Clinical Sciences
Molecular Sciences
Faculty of Medicine
Faculty of Natural Sciences



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