Sensitive light-sheet microscopy in multiwell plates using an AFM cantilever.
Biomedical optics express
Optical Society of America
MetadataShow full item record
Ponjavic, A., Ye, Y., Laue, E., Lee, S., & Klenerman, D. (2018). Sensitive light-sheet microscopy in multiwell plates using an AFM cantilever.. Biomedical optics express, 9 (12), 5863-5880. https://doi.org/10.1364/boe.9.005863
We present a sensitive inverted light sheet microscope, capable of single-molecule fluorescence imaging of cells in 96 multiwell plates. Light sheet microscope designs are often complex and costly, requiring custom-made sample chambers that are incompatible with standard cell culture samples. To overcome this limitation, we have developed single-objective cantilever selective plane illumination (socSPIM), which introduces a light sheet through the objective lens of an inverted microscope using an AFM tip. We demonstrate the effectiveness of this setup by performing 3D imaging of nuclear pore complexes, as well as live whole-cell 3D imaging of lysosomes and super-resolution imaging of the T-cell membrane. The unique advantage offered by socSPIM is the minimal footprint of the cantilever, which allowed us to perform super-resolution reflected light-sheet microscopy by PAINT in 96 multiwell plates, paving the way for high-throughput studies.
The authors acknowledge funding support from Royal Society (UF120277, RP150066); Engineering and Physical Sciences Research Council (EPSRC) (EP/L027631/1); and Wellcome Trust (206291/Z/17/Z). Yu Ye was funded by a Sir Henry Wellcome Fellowship.
EC FP7 CP (277899)
Wellcome Trust (206291/Z/17/Z)
Wellcome Trust (101585/Z/13/Z)
External DOI: https://doi.org/10.1364/boe.9.005863
This record's URL: https://www.repository.cam.ac.uk/handle/1810/286273
Attribution 4.0 International
Licence URL: https://creativecommons.org/licenses/by/4.0/