Table 1.
Overview of fluorescence light-sheet techniques in comparison with epifluorescence, confocal and two-photon microscopy
| Technique | Unique features | Advantages | Disadvantages | Ideal application | References |
|---|---|---|---|---|---|
| Fluorescence microscopy | |||||
| Epifluorescence microscopy | Uniform illumination of the sample | Easy to use, low cost | No optical sectioning | Screening, thin sections | |
| Confocal microscopy | Point illumination (laser focus) and point detection (pinhole) | Optical sectioning, improved resolution (√2) | Slow scanning, low penetration | Medium-thick sections, surface of embryos | Pawley, 2006 |
| Two-photon microscopy | True 3D imaging, infrared laser | Low scattering, excellent penetration, reduced bleaching | Slow scanning, low resolution, high cost, high power beam | Deep penetration in scattering tissue | Helmchen and Denk, 2005 |
|
Fluorescence light-sheet microscopy
|
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| Orthogonal-plane fluorescence optical sectioning (OPFOS) | Single-sided light-sheet illumination | Optical sectioning | Shadows, stripes | Fluorescently stained samples | Voie et al., 1993 |
| High-resolution orthogonal-plane fluorescence optical sectioning (HROPFOS) | Like OPFOS, light-sheet translated | Thin light-sheet of constant thickness | Slow, shadows, stripes | Like OPFOS | Buytaert and Dirckx, 2007 |
| 3D light scanning macrography | Extended depth of field | Large samples | Only surface | Surface scanning of flies | Huber et al., 2001 |
| Thin light-sheet microscope (TLSM) | Light-sheet illuminates tank of medium from the side | Improved contrast in aqueous suspensions | Low resolution | Aquatic microbes | Fuchs et al., 2002 |
| Selective plane illumination microscopy (SPIM) | Single-sided light-sheet illumination, sample rotation about vertical axis | Fast acquisition, low photo-bleaching, good penetration, MVR | Shadows, stripes | Extended time-lapse imaging of millimetre-sized embryos, high speed movies | Huisken et al., 2004 |
| Objective coupled planar illumination (OCPI) microscopy | Single-sided light-sheet illumination, attached to detection lens | Fast stack acquisition, no sample movement, conventional sample | Shadows, stripes | Conventionally prepared samples, fast 3D imaging | Holekamp et al., 2008 |
| Ultramicroscopy | Dual-sided light-sheet illumination, upright detection path | Sealed chamber with clearing solution | No index-matched optics, stripes | Large fixed and cleared specimens | Dodt et al., 2007 |
| Digital scanned laser light-sheet fluorescence microscopy (DSLM) | Like SPIM, light-sheet generated by beam scanning | Like SPIM, uniform, adjustable light-sheet | Like SPIM | Like SPIM | Keller et al., 2008 |
| Multidirectional SPIM (mSPIM) | Like SPIM, pivoting light-sheet, two alternating illumination arms | Like SPIM, reduced stripes, reduced shading | Space constraints | Like SPIM, especially scattering and absorbing tissue | Huisken and Stainier, 2007 |
| Highly inclined and laminated optical sheet (HILO) microscopy | Single lens for oblique light-sheet illumination and detection | Single lens | Very narrow FOV | Single cell microscopy close to the coverslip | Tokunaga et al., 2008 |
| Oblique plane microscopy (OPM) | Like HILO, matching, tilted image plane | Like HILO, extended FOV | Extensive optics | Like HILO | Dunsby, 2008 |