Optical sectioning microscopy with planar or structured illumination.

Department of Biomedical Engineering, Boston University, Boston, Massachusetts, USA.
Nature Methods (Impact Factor: 25.95). 09/2011; 8(10):811-9. DOI: 10.1038/nmeth.1709
Source: PubMed

ABSTRACT A key requirement for performing three-dimensional (3D) imaging using optical microscopes is that they be capable of optical sectioning by distinguishing in-focus signal from out-of-focus background. Common techniques for fluorescence optical sectioning are confocal laser scanning microscopy and two-photon microscopy. But there is increasing interest in alternative optical sectioning techniques, particularly for applications involving high speeds, large fields of view or long-term imaging. In this Review, I examine two such techniques, based on planar illumination or structured illumination. The goal is to describe the advantages and disadvantages of these techniques.

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    • "Techniques like SPIM (Dodt et al., 2007; Keller and Stelzer, 2008), generally called light-sheet microscopy techniques, are gaining popularity because of the high efficiency in accessing volumetric information of the specimen while minimizing photo-bleaching and energy load (Verveer et al., 2007, Holekamp et al., 2008). The advantages of SPIM over confocal was thoroughly discussed (Mertz, 2011). A further development of the SPIM method was proposed by the application of Bessel beams to light sheet microscopy by Planchon et al. (2011). "
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    • "Although the feasibility of a compact confocal microscope for space research has been proven (Beghuin et al., 2005), preference is given to a widefield setup for reasons of simplicity and robustness. Optical sectioning may then be achieved by implementing structured illumination (SIM) (Gustafsson et al., 1999; Mertz, 2011). "
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