We report on the recently emerging (laser) light-sheet-based fluorescence microscopy field (LSFM). The techniques used in this field allow to study and visualize biomedical objects nondestructively in high resolution through virtual optical sectioning with sheets of laser light. Fluorescence originating in the cross-section of the sheet and sample is recorded orthogonally with a camera. In this paper, the first implementation of LSFM to image biomedical tissue in three dimensions—orthogonal-plane fluorescence optical sectioning microscopy (OPFOS)—is discussed. Since then many similar and derived methods have surfaced, (SPIM, ultramicroscopy, HR-OPFOS, mSPIM, DSLM, TSLIM, etc.) which we all briefly discuss. All these optical sectioning methods create images showing histological detail. We illustrate the applicability of LSFM on several specimen types with application in biomedical and life sciences. 1. Introduction Serial (mechanical) histological sectioning (SHS) creates physical slices of fixed, stained, and embedded tissues which are then imaged with an optical microscope in unsurpassed submicrometer resolution. Obtaining these slices is however extremely work intensive, requires physical (one-time and one-directional) slicing and thus destruction of the specimen. A 2D sectional image reveals lots of histologically relevant information, but a data stack and its 3D reconstruction are even more essential for the morphological interpretation of complex structures, because they give additional insight in the anatomy. The SHS method requires semiautomatic to manual image registration to align all recorded 2D slices into order to get realistic 3D reconstructions. Often dedicated image processing of the sections is needed because of the geometrical distortions from the slicing. A valuable alternative to achieve sectional imaging and three-dimensional modeling of anatomic structures can be found in the little known and relatively recent field of microscopy called (laser) light-sheet-based fluorescence microscopy or LSFM. These nondestructive methods generate registered optical sections in real-time through bio(medical) samples ranging from microscopic till macroscopic size. LSFM can reveal both bone and soft tissue at a micrometer resolution, thus showing a large amount of histological detail as well. The first account of the LSFM idea was published by Voie et al. in 1993 and applied to image the inner ear cochlea of guinea pig [1]. Their method was called orthogonal-plane fluorescence optical sectioning (OPFOS) microscopy or tomography. The motivations for
References
[1]
A. H. Voie, D. H. Burns, and F. A. Spelman, “Orthogonal-plane fluorescence optical sectioning: three-dimensional imaging of macroscopic biological specimens,” Journal of Microscopy, vol. 170, no. 3, pp. 229–236, 1993.
[2]
W. Spalteholz, über das Durchsichtigmachen von Menschlichen und Tierischen Pr？paraten, Verlag Hirzel, 1911.
[3]
H. Siedentopf and R. A. Zsigmondy, “Uber sichtbarmachung und gr？？enbestimmung ultramikoskopischer teilchen, mit besonderer anwendung auf goldrubingl？ser,” Annalen der Physik, vol. 315, pp. 1–39, 1903.
[4]
J. A. N. Buytaert and J. J. J. Dirckx, “Tomographic imaging of macroscopic biomedical objects in high resolution and three dimensions using orthogonal-plane fluorescence optical sectioning,” Applied Optics, vol. 48, no. 5, pp. 941–948, 2009.
[5]
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, vol. 305, no. 5686, pp. 1007–1009, 2004.
[6]
J. Huisken and D. Y. R. Stainier, “Even fluorescence excitation by multidirectional selective plane illumination microscopy (mSPIM),” Optics Letters, vol. 32, no. 17, pp. 2608–2610, 2007.
[7]
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,” Journal of Biomedical Optics, vol. 12, no. 1, Article ID 014039, 2007.
[8]
H. U. Dodt, U. Leischner, A. Schierloh et al., “Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain,” Nature Methods, vol. 4, no. 4, pp. 331–336, 2007.
[9]
R. Hofman, J. M. Segenhout, J. A. N. Buytaert, J. J. J. Dirckx, and H. P. Wit, “Morphology and function of Bast's valve: additional insight in its functioning using 3D-reconstruction,” European Archives of Oto-Rhino-Laryngology, vol. 265, no. 2, pp. 153–157, 2008.
[10]
J. Buytaert, E. Descamps, D. Adriaens, and J. Dirckx, “Orthogonal-plane fluorescence optical sectioning : a technique for 3-D imaging of biomedical specimens,” in Microscopy: Science, Technology, Applications and Education, A. Méndez-Vilas and J. Díaz, Eds., pp. 1356–1365, Formatex, 2010.
[11]
P. A. Santi, “Light sheet fluorescence microscopy: a review,” The Journal of Histochemistry and Cytochemistry, vol. 59, no. 2, pp. 129–138, 2011.
[12]
A. H. Voie, “Imaging the intact guinea pig tympanic bulla by orthogonal-plane fluorescence optical sectioning microscopy,” Hearing Research, vol. 171, no. 1-2, pp. 119–128, 2002.
[13]
S. P. Tinling, R. T. Giberson, and R. S. Kullar, “Microwave exposure increases bone demineralization rate independent of temperature,” Journal of Microscopy, vol. 215, no. 3, pp. 230–235, 2004.
[14]
C. F. A. Culling, “Spalteholz technique,” in Handbook of Histopathological and Histochemical Techniques, pp. 550–552, Butterworths, 1974.
[15]
V. Ermolayev, M. Friedrich, R. Nozadze et al., “Ultramicroscopy reveals axonal transport impairments in cortical motor neurons at prion disease,” Biophysical Journal, vol. 96, no. 8, pp. 3390–3398, 2009.
[16]
J. A. N. Buytaert, W. H. M. Salih, M. Dierick, P. Jacobs, and J. J. J. Dirckx, “Realistic 3-D computer model of the gerbil middle ear, featuring accurate morphology of bone and soft tissue structures,” Journal of the Association for Research in Otolaryngology. In press.
[17]
S. Van Wassenbergh, G. Roos, A. Genbrugge et al., “Suction is kid's play: extremely fast suction in newborn seahorses,” Biology Letters, vol. 5, no. 2, pp. 200–203, 2009.
[18]
R. Hofman, J. M. Segenhout, and H. P. Wit, “Three-dimensional reconstruction of the guinea pig inner ear, comparison of OPFOS and light microscopy, applications of 3D reconstruction,” Journal of Microscopy, vol. 233, no. 2, pp. 251–257, 2009.
[19]
J. Lane and Z. A. Ralis, “Changes in dimensions of large cancellous bone specimens during histological preparation as measured on slabs from human femoral heads,” Calcified Tissue International, vol. 35, no. 1, pp. 1–4, 1983.
[20]
A. H. Voie and F. A. Spelman, “Three-dimensional reconstruction of the cochlea from two-dimensional images of optical sections,” Computerized Medical Imaging and Graphics, vol. 19, no. 5, pp. 377–384, 1995.
[21]
E. Fuchs, J. S. Jaffe, R. A. Long, and F. Azam, “Thin laser light sheet microscope for microbial oceanography,” Optics Express, vol. 10, no. 2, pp. 145–154, 2002.
[22]
R. A. Zsigmondy, Properties of Colloids, Nobel lecture in chemistry, Elsevier, Amsterdam, The Netherlands, 1926.
[23]
E. H. K. Stelzer, S. Lindek, S. Albrecht et al., “A new tool for the observation of embryos and other large specimens: confocal theta fluorescence microscopy,” Journal of Microscopy, vol. 179, no. 1, pp. 1–10, 1995.
[24]
S. Lindek and E. H. K. Stelzer, “Confocal theta microscopy and 4Pi-confocal theta microscopy,” in Three-Dimensional Microscopy, SPIE International Society for Optical Engineering, vol. 2184 of Proceedings of the SPIE, p. 188, 1994.
[25]
T. F. Holekamp, D. Turaga, and T. E. Holy, “Fast three-dimensional fluorescence imaging of activity in neural populations by objective-coupled planar illumination microscopy,” Neuron, vol. 57, no. 5, pp. 661–672, 2008.
[26]
C. Dunsby, “Optically sectioned imaging by oblique plane microscopy,” Optics Express, vol. 16, no. 25, pp. 20306–20316, 2008.
[27]
P. J. Keller, A. D. Schmidt, J. Wittbrodt, and E. H. K. Stelzer, “Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy,” Science, vol. 322, no. 5904, pp. 1065–1069, 2008.
[28]
P. A. Santi, S. B. Johnson, M. Hillenbrand, P. Z. GrandPre, T. J. Glass, and J. R. Leger, “Thin-sheet laser imaging microscopy for optical sectioning of thick tissues,” BioTechniques, vol. 46, no. 4, pp. 287–294, 2009.
[29]
J. Mertz and J. Kim, “Scanning light-sheet microscopy in the whole mouse brain with HiLo background rejection,” Journal of Biomedical Optics, vol. 15, no. 1, p. 016027, 2010.
