title: Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates.
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.
authors.affiliation: Photonics Group, Department of Physics, Imperial College London, United Kingdom.
authors.forename: Vincent
authors.initials: V
authors.lastname: Maioli
authors.affiliation: Photonics Group, Department of Physics, Imperial College London, United Kingdom.|MRC Clinical Sciences Centre (CSC), Du Cane Road, London, United Kingdom.|Institute of Clinical Sciences (ICS), Faculty of Medicine, Imperial College London, Du Cane Road, London, United Kingdom.
authors.forename: George
authors.initials: G
authors.lastname: Chennell
authors.affiliation: Photonics Group, Department of Physics, Imperial College London, United Kingdom.
authors.forename: Hugh
authors.initials: H
authors.lastname: Sparks
authors.affiliation: MRC Clinical Sciences Centre (CSC), Du Cane Road, London, United Kingdom.|Institute of Clinical Sciences (ICS), Faculty of Medicine, Imperial College London, Du Cane Road, London, United Kingdom.
authors.forename: Tobia
authors.initials: T
authors.lastname: Lana
authors.affiliation: Photonics Group, Department of Physics, Imperial College London, United Kingdom.
authors.forename: Sunil
authors.initials: S
authors.lastname: Kumar
authors.affiliation: MRC Clinical Sciences Centre (CSC), Du Cane Road, London, United Kingdom.|Institute of Clinical Sciences (ICS), Faculty of Medicine, Imperial College London, Du Cane Road, London, United Kingdom.
authors.forename: David
authors.initials: D
authors.lastname: Carling
authors.affiliation: MRC Clinical Sciences Centre (CSC), Du Cane Road, London, United Kingdom.|Institute of Clinical Sciences (ICS), Faculty of Medicine, Imperial College London, Du Cane Road, London, United Kingdom.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Sardini
authors.affiliation: Photonics Group, Department of Physics, Imperial College London, United Kingdom.|Centre for Pathology, Faculty of Medicine, Imperial College London, United Kingdom.
authors.forename: Chris
authors.initials: C
authors.lastname: Dunsby
mesh_terms: D015374:Biosensing Techniques; D031541:Fluorescence Resonance Energy Transfer; D005947:Glucose / Q000032:analysis*; D057809:HEK293 Cells; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D018874:Spheroids, Cellular
chemical_list: D005947:Glucose