Sensitive light-sheet microscopy in multiwell plates using an AFM cantilever.

Change log
Ponjavic, Aleks 
Lee, Steven F 

We present a sensitive inverted light sheet microscope, capable of single-molecule fluorescence imaging of cells in 96-well 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 microscopy (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-well plates, paving the way for high-throughput studies.

0601 Biochemistry and Cell Biology, 0299 Other Physical Sciences, Bioengineering, Biotechnology, Generic Health Relevance
Journal Title
Biomed Opt Express
Conference Name
Journal ISSN
Volume Title
The Optical Society
Engineering and Physical Sciences Research Council (EP/L027631/1)
Medical Research Council (MR/M010082/1)
European Commission (277899)
Medical Research Council (MR/P019471/1)
Wellcome Trust (206291/Z/17/Z)
Wellcome Trust (101585/Z/13/Z)
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.