Light-sheet microscopy
has revolutionized bioimaging by enabling
approximately an order of magnitude reduction in specimen irradiance
compared to confocal imaging. Here, we introduce a light-sheet imaging
system that enables an additional order of magnitude reduction in
specimen irradiance by operating at the Poisson limit. To operate
at this limit, we integrated classical illumination with single-photon
detection and wavelet-based image reconstruction. This integration
enabled brightness quantification and object recognition from fewer
than one detected photon per image pixel, corresponding to more than
10-fold lower irradiance levels than modern systems. We demonstrate
how such photon-sparse imaging can eradicate photobleaching and enable
both dim and bright object imaging, thus, further enhancing the related
gains of light-sheet microscopy.
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