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. 2021 Sep 27;29(20):32691-32699.
doi: 10.1364/OE.439338.

Incoherent superposition of polychromatic light enables single-shot nondiffracting light-sheet microscopy

Vahid Ebrahimi, Jialei Tang, Kyu Young Han

Incoherent superposition of polychromatic light enables single-shot nondiffracting light-sheet microscopy

Vahid Ebrahimi et al. Opt Express. .

Abstract

We demonstrate single-shot nondiffracting light-sheet microscopy by the incoherent superposition of dispersed polychromatic light sources. We characterized our technique by generating a Bessel light-sheet with a supercontinuum light-source and a C-light-sheet using a diode laser, and demonstrated its applicability to fluorescence microscopy. We emphasize that our method is easily implementable and compatible with the requirements of high-resolution microscopy.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Fig. 1.
Fig. 1.
Working principle of single-shot nondiffracting light-sheet generation by dispersed 1D coherent beams. Incoherent superposition of fields resulting from each different wavelength generates nondiffracting light-sheets such as Bessel light-sheet (top) or C-light-sheet (bottom). FT denotes Fourier transform. Scale bars, 50 μm.
Fig. 2.
Fig. 2.
Experimental scheme for creating 1D coherent beams using polychromatic light. A reflective diffraction grating followed by a cylindrical lens disperses light and focuses them at an annulus placed at the back focal plane of an objective lens.
Fig. 3.
Fig. 3.
Experimental setup of the single-shot nondiffracting light-sheet microscope. A diode laser (top) or a supercontinuum light source (bottom) was used as an excitation source. CAM1-2, cameras; CL, cylindrical lens; L1-8, lenses; SMF, single-mode fiber. a and b denote inner and outer radius, respectively.
Fig. 4.
Fig. 4.
Light-sheet intensity profiles generated by a supercontinuum light source. (a) Bessel beam. (b) Gaussian light-sheet. (c) Bessel light-sheet. (d) Line profiles of Gaussian (blue) and Bessel (red) light-sheets along the x-axis (top) and z-axis (bottom). Scale bars, 100 μm (white) and 1 mm (yellow).
Fig. 5.
Fig. 5.
Single-shot C-light-sheet generation using a diode laser. The beam covers a range of 0.43-0.85mm at the BFP. (a) Intensity profiles of single-shot C-light-sheet. (b) Spectrum of the diode laser. Line profiles in the x-axis (c) and z-axis (d) for the Gaussian light-sheet (blue) and C-light-sheet (red). Scale bars, 25 μm (white) and 400 μm (yellow).
Fig. 6.
Fig. 6.
Fluorescence images of 1 μm beads in 3D hydrogels measured by Gaussian light-sheet (a) and single-shot C-light-sheet microscopy (b-d). The detection objectives were 4×/0.13 for (a, b) and 50×/0.8 for (c, d). All images were maximum intensity projected. Scale bars, 10 μm (white) and 200 μm (yellow).
Fig. 7.
Fig. 7.
Rat-tail collagen labeled with Atto647N imaged by a Gaussian light-sheet (left) and a single-shot C-light-sheet (right). The detection objective lens was 20×/0.5. Scale bar, 150 μm.
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