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Review
. 2018 May 14;26(10):13122-13147.
doi: 10.1364/OE.26.013122.

Light sheet approaches for improved precision in 3D localization-based super-resolution imaging in mammalian cells [Invited]

Anna-Karin Gustavsson, Petar N Petrov, W E Moerner
Review

Light sheet approaches for improved precision in 3D localization-based super-resolution imaging in mammalian cells [Invited]

Anna-Karin Gustavsson et al. Opt Express. .

Abstract

The development of imaging techniques beyond the diffraction limit has paved the way for detailed studies of nanostructures and molecular mechanisms in biological systems. Imaging thicker samples, such as mammalian cells and tissue, in all three dimensions, is challenging due to increased background and volumes to image. Light sheet illumination is a method that allows for selective irradiation of the image plane, and its inherent optical sectioning capability allows for imaging of biological samples with reduced background, photobleaching, and photodamage. In this review, we discuss the advantage of combining single-molecule imaging with light sheet illumination. We begin by describing the principles of single-molecule localization microscopy and of light sheet illumination. Finally, we present examples of designs that successfully have married single-molecule super-resolution imaging with light sheet illumination for improved precision in mammalian cells.

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Figures

Fig. 1
Fig. 1
Fundamentals of localization-based super-resolution microscopy. The noisy image (a) of an isolated emitter can be fitted to a model function such as a Gaussian (b) to estimate the center position. The distribution of center position estimates (c) is much narrower than the point spread function. A diffraction-limited image with all molecules fluorescing simultaneously (d) can be super-resolved by sequentially localizing spatially-isolated molecules (e) to produce a reconstruction (f) which shows fine structural details. Localization precision σµx for various signal photons N and background photons per pixel β is plotted in (g), based on Eq. (1) for a 160 nm pixel size and 250 nm diffraction-limited spot size.
Fig. 2
Fig. 2
Experimental demonstrations of engineered point spread functions (PSFs) used for 3D localization microscopy. The arrows (right) represent the applicable axial ranges of the different PSFs, and the range over which the PSFs were imaged. (a) Astigmatic [37]. Scale bar ~0.5 µm. Reprinted from [37]. Reprinted with permission from AAAS. (b) Phase ramp [50]. Reprinted with kind permission from Springer. (c) Double-helix [41]. Scale bar is 2 µm. Reprinted with permission from Ref [41]. (d) Accelerating beam [52]. Scale bar is 1 µm. Reprinted by permission from Macmillan Publishers Ltd: Nature Photonics [52], copyright (2014). (e) Corkscrew [51]. Reprinted with permission from [51]. (f), (g) Tetrapods [54]. Scale bars are 2 µm and 5 µm in (f) and (g), respectively. Reprinted from [54] with permission from the American Chemical Society (http://pubs.acs.org/doi/abs/10.1021%2Facs.nanolett.5b01396). (h) Schematic of the optical design used for PSF engineering when implemented using a reflective element for phase modulation. Figure adapted from Ref [75]. with permission from the Royal Society of Chemistry.
Fig. 3
Fig. 3
Comparison between epi-, confocal, total internal reflection fluorescence (TIRF), and light sheet illumination. Epi- and confocal illumination provides no axial confinement of the irradiation, which causes unnecessary background (epi), photobleaching, and photodamage. Confocal illumination improves contrast by blocking out-of-focus light but requires scanning to build up an image. TIRF provides excellent optical sectioning, but is limited to imaging within a few hundred nm from the coverslip. Light sheet provides an elegant means to illuminate only the sample plane that is imaged, which reduces both background, photobleaching, and photodamage.
Fig. 4
Fig. 4
Simulated beam profiles using the sample-oriented axis convention with x the propagation direction of the beam and z the optical axis of the collection objective. Gaussian yz (a) and xz (b), Bessel yz (c) and xz (d), and Airy yz (e) and xz (f) are shown. The yz profiles of four beams are compared: Gaussian beam formed by cylindrical lens (g), scanned Gaussian beam (h), scanned single-photon Bessel beam (i), and scanned two-photon Bessel beam (j). All scale bars are 10 µm. Each image is scaled to span the color map on the right.
Fig. 5
Fig. 5
Schematics of different designs that can be used for light sheet single-molecule super-resolution imaging with improved contrast. HILO: highly inclined and laminated optical sheet [174]; IML-SPIM: individual molecule localization with selective-plane illumination microscopy [175]; RLSM: reflected light sheet microscopy [176]; LSBM: light-sheet Bayesian microscopy [177]; LLS: lattice light-sheet [154]; soSPIM: single-objective SPIM [178]; SO-LSM: single-objective light-sheet microscopy [141]; TILT3D: tilted light sheet microscopy with 3D PSFs [49]. Schematics are not to scale.
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