High-speed and high-content light-sheet fluorescence microscopy

Light-sheet fluorescence microscopy (LSFM) provides low out-of-plane photobleaching and phototoxicity, but usually requires two microscope objective lenses orientated at 90? to one another - one for fluorescence excitation and one for fluorescence detection - making it harder to image samples prepared using conventional mounting methods. Oblique plane microscopy (OPM) is a type of LSFM that has been developed in our laboratory and uses a single high numerical aperture microscope objective to provide both fluorescence excitation and detection whilst maintaining the advantages of LSFM, enabling it to provide high-speed 3D imaging for a range of applications on a conventional fluorescence microscope frame. The speed of OPM imaging can be applied to image a single sample at video volumetric imaging rates. It can also be used to enable higher throughput and time-lapse 3D imaging of arrays of samples arrayed in multi-well plates. This talk will present examples of the application of OPM for high-speed 3D imaging of isolated cardiomyocytes and also examples where the system is being applied to study arrays of multicellular spheroids and organoids in 3D over multiple conditions and over time.

Hugh Sparks

Research Associate

Imperial College London

Dr Hugh Sparks received a masters degree in physics from The University of Manchester and a PhD in physics from Imperial College London. During the PhD, they developed novel time-resolved fluorescence imaging endoscope technologies for biomedical applications. They then worked in Erik Sahai’s Tumor Cell Biology Lab at The Francis Crick Institute to investigate the application of time-resolved fluorescence imaging endoscopy to pre-clinical imaging of drug-target engagement during chemotherapy. Currently they are working as a Cancer Research UK (CRUK) funded postdoctoral researcher in Chris Dunsby’s and Paul French’s photonics labs at Imperial College London and as a visiting researcher at The Institute of Cancer Research (IRC) and The Francis Crick Institute. The primary scope of the work is to develop new tools to enable higher throughput and time-lapse 3D imaging of samples arrayed in multi-well plates for cancer research.

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High-speed and high-content light-sheet fluorescence microscopy
Open to view video.  |   Closed captions available
Open to view video.  |   Closed captions available Light-sheet fluorescence microscopy (LSFM) provides low out-of-plane photobleaching and phototoxicity, but usually requires two microscope objective lenses orientated at 90° to one another – one for fluorescence excitation and one for fluorescence detection – making it harder to image samples prepared using conventional mounting methods. Oblique plane microscopy (OPM) is a type of LSFM that has been developed in our laboratory and uses a single high numerical aperture microscope objective to provide both fluorescence excitation and detection whilst maintaining the advantages of LSFM, enabling it to provide high-speed 3D imaging for a range of applications on a conventional fluorescence microscope frame. The speed of OPM imaging can be applied to image a single sample at video volumetric imaging rates. It can also be used to enable higher throughput and time-lapse 3D imaging of arrays of samples arrayed in multi-well plates. This talk will present examples of the application of OPM for high-speed 3D imaging of isolated cardiomyocytes and also examples where the system is being applied to study arrays of multicellular spheroids and organoids in 3D over multiple conditions and over time.
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