We recently demonstrated that Microscopy with Self-Reconstructing Beams (MISERB) increases both image quality and penetration depth of illumination beams in strongly scattering media. Based on the concept of line scanned light-sheet microscopy, we present an add-on module to a standard inverted microscope using a scanned beam that is shaped in phase and amplitude by a spatial light modulator. We explain technical details of the setup as well as of the holograms for the creation, positioning and scaling of static light-sheets, Gaussian beams and Bessel beams. The comparison of images from identical sample areas illuminated by different beams allows a precise assessment of the interconnection between beam shape and image quality. The superior propagation ability of Bessel beams through inhomogeneous media is demonstrated by measurements on various scattering media.

© 2010 OSA

1. F. Pampaloni, E. G. Reynaud, and E. H. K. Stelzer, “The third dimension bridges the gap between cell culture and live tissue,” Nat. Rev. Mol. Cell Biol. 8(10), 839–845 (2007). [CrossRef] [PubMed]
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3. A. H. Voie, D. H. Burns, and F. A. Spelman, “Orthogonal-plane fluorescence optical sectioning: three-dimensional imaging of macroscopic biological specimens,” J. Microsc. 170(Pt 3), 229–236 (1993). [CrossRef] [PubMed]
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10. W. T. Welford, “Use of Annular Apertures to Increase Focal Depth,” J. Opt. Soc. Am. 50(8), 749–753 (1960). [CrossRef]
11. J. W. Y. Lit and R. Tremblay, “Focal Depth of a Transmitting Axicon,” J. Opt. Soc. Am. 63(4), 445–449 (1973). [CrossRef]
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13. Z. Bouchal, J. Wagner, and M. Chlup, “Self-reconstruction of a distorted nondiffracting beam,” Opt. Commun. 151(4-6), 207–211 (1998). [CrossRef]
14. V. Garcés-Chávez, D. McGloin, H. Melville, W. Sibbett, and K. Dholakia, “Simultaneous micromanipulation in multiple planes using a self-reconstructing light beam,” Nature 419(6903), 145–147 (2002). [CrossRef] [PubMed]
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18. R. D. L. Hanes, M. C. Jenkins, and S. U. Egelhaaf, “Combined holographic-mechanical optical tweezers: construction, optimization, and calibration,” Rev. Sci. Instrum. 80(8), 083703 (2009). [CrossRef] [PubMed]
19. C. W. McCutchen, “Generalized Aperture and the Three-Dimensional Diffraction Image,” J. Opt. Soc. Am. 54(2), 240–244 (1964). [CrossRef]
20. Y. Roichman and D. G. Grier, “Projecting extended optical traps with shape-phase holography,” Opt. Lett. 31(11), 1675–1677 (2006). [CrossRef] [PubMed]
21. A. Jesacher, C. Maurer, A. Schwaighofer, S. Bernet, and M. Ritsch-Marte, “Near-perfect hologram reconstruction with a spatial light modulator,” Opt. Express 16(4), 2597–2603 (2008). [CrossRef] [PubMed]
22. M. Agour, E. Kolenovic, C. Falldorf, and C. von Kopylow, “Suppression of higher diffraction orders and intensity improvement of optically reconstructed holograms from a spatial light modulator,” J. Opt. A 11, 105405 (2009). [CrossRef]
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Appl. Opt.

• E. G. van Putten, I. M. Vellekoop, and A. P. Mosk, “Spatial amplitude and phase modulation using commercial twisted nematic LCDs,” Appl. Opt. 47(12), 2076–2081 (2008). [CrossRef] [PubMed]

Development

• J. Huisken and D. Y. Stainier, “Selective plane illumination microscopy techniques in developmental biology,” Development 136(12), 1963–1975 (2009). [CrossRef] [PubMed]

HFSP J

• E. G. Reynaud, U. Krzic, K. Greger, and E. H. K. Stelzer, “Light sheet-based fluorescence microscopy: more dimensions, more photons, and less photodamage,” HFSP J 2(5), 266–275 (2008). [CrossRef]

J. Biomed. Opt.

• J. A. N. Buytaert and J. J. J. Dirckx, “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). [CrossRef] [PubMed]

J. Microsc.

• A. H. Voie, D. H. Burns, and F. A. Spelman, “Orthogonal-plane fluorescence optical sectioning: three-dimensional imaging of macroscopic biological specimens,” J. Microsc. 170(Pt 3), 229–236 (1993). [CrossRef] [PubMed]

J. Opt. A

• M. Agour, E. Kolenovic, C. Falldorf, and C. von Kopylow, “Suppression of higher diffraction orders and intensity improvement of optically reconstructed holograms from a spatial light modulator,” J. Opt. A 11, 105405 (2009). [CrossRef]

J. Opt. Soc. Am.

• C. W. McCutchen, “Generalized Aperture and the Three-Dimensional Diffraction Image,” J. Opt. Soc. Am. 54(2), 240–244 (1964). [CrossRef]
• W. T. Welford, “Use of Annular Apertures to Increase Focal Depth,” J. Opt. Soc. Am. 50(8), 749–753 (1960). [CrossRef]
• J. W. Y. Lit and R. Tremblay, “Focal Depth of a Transmitting Axicon,” J. Opt. Soc. Am. 63(4), 445–449 (1973). [CrossRef]

Nat. Methods

• V. Ntziachristos, “Going deeper than microscopy: the optical imaging frontier in biology,” Nat. Methods 7(8), 603–614 (2010). [CrossRef] [PubMed]
• H. U. Dodt, U. Leischner, A. Schierloh, N. Jährling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgänsberger, and K. Becker, “Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain,” Nat. Methods 4(4), 331–336 (2007). [CrossRef] [PubMed]

Nat. Photonics

• F. O. Fahrbach, P. Simon, and A. Rohrbach, “Microscopy with self-reconstructing beams,” Nat. Photonics (2010), doi: . [CrossRef]

Nat. Rev. Mol. Cell Biol.

• F. Pampaloni, E. G. Reynaud, and E. H. K. Stelzer, “The third dimension bridges the gap between cell culture and live tissue,” Nat. Rev. Mol. Cell Biol. 8(10), 839–845 (2007). [CrossRef] [PubMed]

Nature

• V. Garcés-Chávez, D. McGloin, H. Melville, W. Sibbett, and K. Dholakia, “Simultaneous micromanipulation in multiple planes using a self-reconstructing light beam,” Nature 419(6903), 145–147 (2002). [CrossRef] [PubMed]

Opt. Commun.

• Z. Bouchal, J. Wagner, and M. Chlup, “Self-reconstruction of a distorted nondiffracting beam,” Opt. Commun. 151(4-6), 207–211 (1998). [CrossRef]

Opt. Express

• A. Jesacher, C. Maurer, A. Schwaighofer, S. Bernet, and M. Ritsch-Marte, “Near-perfect hologram reconstruction with a spatial light modulator,” Opt. Express 16(4), 2597–2603 (2008). [CrossRef] [PubMed]

Opt. Lett.

• Y. Roichman and D. G. Grier, “Projecting extended optical traps with shape-phase holography,” Opt. Lett. 31(11), 1675–1677 (2006). [CrossRef] [PubMed]
• J. Huisken and D. Y. R. Stainier, “Even fluorescence excitation by multidirectional selective plane illumination microscopy (mSPIM),” Opt. Lett. 32(17), 2608–2610 (2007). [CrossRef] [PubMed]
• A. Rohrbach, “Artifacts resulting from imaging in scattering media: a theoretical prediction,” Opt. Lett. 34(19), 3041–3043 (2009). [CrossRef] [PubMed]

Phys. Rev. Lett.

• J. Durnin, J. J. Miceli, and J. H. Eberly, “Diffraction-free beams,” Phys. Rev. Lett. 58(15), 1499–1501 (1987). [CrossRef] [PubMed]

Rev. Sci. Instrum.

• R. D. L. Hanes, M. C. Jenkins, and S. U. Egelhaaf, “Combined holographic-mechanical optical tweezers: construction, optimization, and calibration,” Rev. Sci. Instrum. 80(8), 083703 (2009). [CrossRef] [PubMed]

Science

• P. J. Keller, A. D. Schmidt, J. Wittbrodt, and E. H. Stelzer, “Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy,” Science 322(5904), 1065–1069 (2008). [CrossRef] [PubMed]
• J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science 305(5686), 1007–1009 (2004). [CrossRef] [PubMed]

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