Phasor-based hyperspectral snapshot microscopy allows fast imaging of live three-dimensional tissues for biomedical applications

摘要

a, b Conventional hyperspectral imaging methods rely on slow spatial or spectral scanning. c Faster snapshot methods exist but often compromise spatial and/or spectral resolution and can be computationally demanding. d In conventional phasor-based hyperspectral imaging, an emission spectrum is acquired in each pixel/voxel that is then computationally transformed to the spectral phasor plot for analysis by a sine/cosine transformation. e Instead, the sine/cosine filter approach to hyperspectral imaging directly transforms the fluorescence emission to spectral phasor coordinates in hardware, no spectral detector and image data post-processing are required. f The resulting substantial improvement in image acquisition and data processing speed allows the generation of 4D data (x, y, z, λ) in seconds taking full advantage of the high light throughput and minimal photobleaching of light sheet microscopy.