Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2014 Sep 5;5(10):3434-42.
doi: 10.1364/BOE.5.003434. eCollection 2014 Oct 1.

A compact Airy beam light sheet microscope with a tilted cylindrical lens

Zhengyi Yang  1 , Martynas Prokopas  1 , Jonathan Nylk  2 , Clara Coll-Lladó  3 , Frank J Gunn-Moore  4 , David E K Ferrier  3 , Tom Vettenburg  1 , Kishan Dholakia  1
Affiliations

A compact Airy beam light sheet microscope with a tilted cylindrical lens

Zhengyi Yang et al. Biomed Opt Express. .

Abstract

Light-sheet imaging is rapidly gaining importance for imaging intact biological specimens. Many of the latest innovations rely on the propagation-invariant Bessel or Airy beams to form an extended light sheet to provide high resolution across a large field of view. Shaping light to realize propagation-invariant beams often relies on complex programming of spatial light modulators or specialized, custom made, optical elements. Here we present a straightforward and low-cost modification to the traditional light-sheet setup, based on the open-access light-sheet microscope OpenSPIM, to achieve Airy light-sheet illumination. This brings wide field single-photon light-sheet imaging to a broader range of endusers. Fluorescent microspheres embedded in agarose and a zebrafish larva were imaged to demonstrate how such a microscope can have a minimal footprint and cost without compromising on imaging quality.

Keywords: (070.0070) Fourier optics and signal processing; (110.0180) Microscopy; (110.1758) Computational imaging; (110.4850) Optical transfer functions; (110.7348) Wavefront encoding; (140.3300) Laser beam shaping; (170.0170) Medical optics and biotechnology; (180.6900) Three-dimensional microscopy; (220.1000) Aberration compensation.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
Schematic of the open-access Airy light-sheet microscope. (a) Experimental setup with tilted cylindrical lens (CL). (b) shows the projection of measured Airy light sheet. (c) The red line shows the beam profile of a cross section on (b). The blue line indicates the corresponding Airy beam profile from fitted model.
Fig. 2
Fig. 2
Influence of the cylindrical lens tilt angle. (a) focal length, (b) optical axis displacement, (c) cubic and, (d) higher order modulation residual, as a function of the lens angle. The line colors correspond to the wavelengths 405nm, 488nm, 532nm, 561nm, and 633nm. At the wavelength of 532nm used in the experiments, the α value is 1.21 at 35°, 4.13 at 40°, and 11.24 at 45°, covering the values useful for Airy beam light sheet microscopy. Almost no higher order terms are present as can be seen from (d). The residual phase modulation has a standard deviation of only 0.026λ, 0.006λ, and 0.061λ, respectively. The corresponding focal lengths are 26mm, 22mm, and 18mm. The axis position shifts by 1.6mm, 1.9mm, and, 2.3mm. The focus position at a wavelength of 488nm differs by less than 1%. As a result, the optics do not need to be adjusted for minor changes in wavelength.
Fig. 3
Fig. 3
Vertical projection of a sample with fluorescent microspheres (∅ = 600nm), before deconvolution (a), and after deconvolution (b). Although before deconvolution the fluorescent microspheres appear blurred in the axial dimension, z, the pattern is relatively independent of the horizontal coordinate, x. Deconvolution using the light sheet model yields comparable resolution throughout the FOV.
Fig. 4
Fig. 4
Light-sheet microscopy scan of the musculature of a zebrafish larva. Two distant sections along the light sheet propagation axis (x) are shown at x = +45 μm (a) and x = −10 μm (c). Two-dimensional sections are shown at y = −40 μm (b) and at z = −10 μm (d). The thickness of each section is 7.4 μm. The transverse view of the musculature (b) shows the diameter of individual muscle fibers, which vary in size. In the longitudinal views of the musculature (c, d), a fragment of the V-shaped myosepta separating two muscle blocks can be appreciated: in (c) the myosepta (dark diagonal line) extends from (y = −20 μm, z = 20 μm) to (y = 10 μm, z = 50 μm), and in (d) extends from (x = 70 μm, z = 15 μm) to (x = 40 μm, z = 45 μm).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Huisken J., Swoger J., Del Bene F., Wittbrodt J., Stelzer E. H. K., “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science 305(5686), 1007–1009 (2004).10.1126/science.1100035 - DOI - PubMed
    1. Engelbrecht C. J., Stelzer E. H. K., “Resolution enhancement in a light-sheet-based microscope (SPIM),” Opt. Lett. 31(10), 1477–1479 (2006).10.1364/OL.31.001477 - DOI - PubMed
    1. Huisken J., Stainier D. Y. R., “Even fluorescence excitation by multidirectional selective plane illumination microscopy (mSPIM),” Opt. Lett. 32(17), 2608–2610 (2007).10.1364/OL.32.002608 - DOI - PubMed
    1. Tomer R., Khairy K., Amat F., Keller P. J., “Quantitative high-speed imaging of entire developing embryos with simultaneous multiview light-sheet microscopy,” Nat. Methods 9, 755–763 (2012).10.1038/nmeth.2062 - DOI - PubMed
    1. Buytaert J. A. N., Dirckx J. J. J., “Design and quantitative resolution measurements of an optical virtual sectioning three-dimensional imaging technique for biomedical specimens, featuring two-micrometer slicing resolution,” J. Biomed. Opt. 12(1), 014039 (2007).10.1117/1.2671712 - DOI - PubMed
  NODES
3d 1
coding 1
Experiments 1
HOME 2
innovation 1
Javascript 1
mac 2
os 51
text 12
twitter 2
Users 1
web 4