Abstract
We demonstrate the first planar Airy light-sheet microscope. Fluorescence light-sheet microscopy has become the method of choice to study large biological samples with cellular or sub-cellular resolution. The propagation-invariant Airy beam enables a ten-fold increase in field-of-view with single-photon excitation; however, the characteristic asymmetry of the light-sheet limits its potential for multi-photon excitation. Here we show how a planar light-sheet can be formed from the curved propagation-invariant Airy beam. The resulting symmetric light sheet excites two-photon fluorescence uniformly across an extended field-of-view without the need for deconvolution. We demonstrate the method for rapid two-photon imaging of large volumes of neuronal tissue.
Highlights
Fluorescence light-sheet microscopy has found rapid adoption in developmental biology and the neurosciences 8
Light-sheets formed by such beams do have a transversal structure of sidelobes that would lead to poor axial resolution, unless the fluorescence excited by the side lobes is blocked 6, or the main lobe is singled out using structured illumination or two-photon Bessel beam excitation 16
A two-photon excitation (2PE) Airy light-sheet was produced by introducing a cubic phase mask in the illumination path of an inverted light-sheet microscope
Summary
Fluorescence light-sheet microscopy has found rapid adoption in developmental biology and the neurosciences 8. Diffraction limits the field-ofview (FOV) in which the light-sheet illumination can be confined to a sufficiently thin section. This leads to a loss in contrast and axial resolution in all but the center of the FOV, while ineffectively exciting fluorescence elsewhere. Tiling multiple acquisitions 7, or swiping the focus of the light-sheet across the FOV may Abbreviations: 2PE, two-photon excitation; 3D, threedimensional; FOV, field-of-view; FWHM, full-width at halfmaximum; EGFP, enhanced green fluorescent protein; NA, numerical aperture; PDMS, Polydimethylsiloxane. Light-sheets formed by such beams do have a transversal structure of sidelobes that would lead to poor axial resolution, unless the fluorescence excited by the side lobes is blocked 6, or the main lobe is singled out using structured illumination or two-photon Bessel beam excitation 16
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