Light sheet fluorescence microscopy

Last updated

The principle setup of a light sheet fluorescence microscope. Spim prinziple en.svg
The principle setup of a light sheet fluorescence microscope.

Light sheet fluorescence microscopy (LSFM) is a fluorescence microscopy technique with an intermediate-to-high [1] optical resolution, but good optical sectioning capabilities and high speed. In contrast to epifluorescence microscopy only a thin slice (usually a few hundred nanometers to a few micrometers) of the sample is illuminated perpendicularly to the direction of observation. For illumination, a laser light-sheet is used, i.e. a laser beam which is focused only in one direction (e.g. using a cylindrical lens). A second method uses a circular beam scanned in one direction to create the lightsheet. As only the actually observed section is illuminated, this method reduces the photodamage and stress induced on a living sample. Also the good optical sectioning capability reduces the background signal and thus creates images with higher contrast, comparable to confocal microscopy. Because light sheet fluorescence microscopy scans samples by using a plane of light instead of a point (as in confocal microscopy), it can acquire images at speeds 100 to 1,000 times faster than those offered by point-scanning methods.

Contents

Comparison of different microscopy illumination modalities (LSFM: light sheet fluorescence microscopy, WF: widefield microscopy, CF: confocal microscopy). Light sheet fluorescence microscopy combines good z-sectioning (as confocal) and illuminates only the observed plane Lsfm lightsheetinsample.svg
Comparison of different microscopy illumination modalities (LSFM: light sheet fluorescence microscopy, WF: widefield microscopy, CF: confocal microscopy). Light sheet fluorescence microscopy combines good z-sectioning (as confocal) and illuminates only the observed plane

This method is used in cell biology [2] and for microscopy of intact, often chemically cleared, organs, embryos, and organisms. [3]

Starting in 1994, light sheet fluorescence microscopy was developed as orthogonal plane fluorescence optical sectioning microscopy or tomography (OPFOS) [4] mainly for large samples and later as the selective/single plane illumination microscopy (SPIM) also with sub-cellular resolution. [5] This introduced an illumination scheme into fluorescence microscopy, which has already been used successfully for dark field microscopy under the name ultramicroscopy. [6]

Setup

Basic setup

Illustration of different light sheet fluorescence microscope implementations. See text for details. Legend: CAM=camera, TL=tube lens, F=filter, DO=detection objective, S=sample, SC=sample chamber, PO=projection objective, CL=cylindrical lens, SM=scanning mirror Lsfm methods.svg
Illustration of different light sheet fluorescence microscope implementations. See text for details. Legend: CAM=camera, TL=tube lens, F=filter, DO=detection objective, S=sample, SC=sample chamber, PO=projection objective, CL=cylindrical lens, SM=scanning mirror

In this type of microscopy, [7] the illumination is done perpendicularly to the direction of observation (see schematic image at the top of the article). The expanded beam of a laser is focused in only one direction by a cylindrical lens, or by a combination of a cylindrical lens and a microscope objective as the latter is available in better optical quality and with higher numerical aperture than the first. This way a thin sheet of light or lightsheet is created in the focal region that can be used to excite fluorescence only in a thin slice (usually a few micrometers thin) of the sample.

The fluorescence light emitted from the lightsheet is then collected perpendicularly with a standard microscope objective and projected onto an imaging sensor (usually a CCD, electron-multiplying CCD or CMOS camera). In order to let enough space for the excitation optics/lightsheet an observation objective with high working distance is used. In most light sheet fluorescence microscopes the detection objective and sometimes also the excitation objective are fully immersed in the sample buffer, so usually the sample and excitation/detection optics are embedded into a buffer-filled sample chamber, which can also be used to control the environmental conditions (temperature, carbon dioxide level ...) during the measurement. The sample mounting in light sheet fluorescence microscopy is described below in more detail.

As both the excitation lightsheet and the focal plane of the detection optics have to coincide to form an image, focusing different parts of the sample can not be done by translating the detection objective, but usually the whole sample is translated and rotated instead.

Extensions of the basic idea

In recent years, several extensions to this scheme have been developed:

Sample mounting

Different types of sample mounting for light sheet fluorescence microscopy: embryo embedded in hanging gel cylinder, plant growing in supported gel cylinder, adherent cells on glass, liquid sample in a sample bag Lsfm samplemounting.svg
Different types of sample mounting for light sheet fluorescence microscopy: embryo embedded in hanging gel cylinder, plant growing in supported gel cylinder, adherent cells on glass, liquid sample in a sample bag

The separation of the illumination and detection beampaths in light sheet fluorescence microscopy (except in oblique plane microscopy) creates a need for specialized sample mounting methods. To date most light sheet fluorescence microscopes are built in such a way that the illumination and detection beampath lie in a horizontal plane (see illustrations above), thus the sample is usually hanging from the top into the sample chamber or is resting on a vertical support inside the sample chamber. Several methods have been developed to mount all sorts of samples:

Some light sheet fluorescence microscopes have been developed where the sample is mounted as in standard microscopy (e.g. cells grow horizontally on the bottom of a petri dish) and the excitation and detection optics are constructed in an upright plane from above. This also allows combining a light sheet fluorescence microscope with a standard inverted microscope and avoids the requirement for specialized sample mounting procedures. [20] [30] [31] [32]

Image properties

Typical imaging modes

Most light sheet fluorescence microscopes are used to produce 3D images of the sample by moving the sample through the image plane. If the sample is larger than the field of view of the image sensor, the sample also has to be shifted laterally. An alternative approach is to move the image plane through the sample to create the image stack. [32]

Long experiments can be carried out, for example with stacks recorded every 10 sec–10 min over the timespan of days. This allows study of changes over time in 3D, or so-called 4D microscopy.

After the image acquisition the different image stacks are registered to form one single 3D dataset. Multiple views of the sample can be collected, either by interchanging the roles of the objectives [32] or by rotating the sample. [8] Having multiple views can yield more information than a single stack; for example occlusion of some parts of the sample may be overcome. Multiple views also improves 3D image resolution by overcoming poor axial resolution as described below.

Some studies also use a selective plane illumination microscope to image only one slice of the sample, but at much higher temporal resolution. This allows e.g. to observe the beating heart of a zebra fish embryo in real-time. [33] Together with fast translation stages for the sample a high-speed 3D particle tracking has been implemented. [34]

Power of resolution

The lateral resolution of a selective plane illumination microscope is comparable to that of a standard (epi) fluorescence microscope, as it is determined fully by the detection objective and the wavelength of the detected light (see Abbe limit). E.g. for detection in the green spectral region around 525 nm, a resolution of 250–500 nm can be reached. [7] The axial resolution is worse than the lateral (about a factor of 4), but it can be improved by using a thinner lightsheet in which case nearly isotropic resolution is possible. [20] Thinner light sheets are either thin only in a small region (for Gaussian beams) or else specialized beam profiles such as Bessel beams must be used (besides added complexity, such schemes add side lobes which can be detrimental). [13] Alternatively, isotropic resolution can be achieved by computationally combining 3D image stacks taken from the same sample under different angles. Then the depth-resolution information lacking in one stack is supplied from another stack; for example with two orthogonal stacks the (poor-resolution) axial direction in one stack is a (high-resolution) lateral direction in the other stack.

The lateral resolution of light sheet fluorescence microscopy can be improved beyond the Abbe limit, by using super resolution microscopy techniques, e.g. with using the fact, that single fluorophores can be located with much higher spatial precision than the nominal resolution of the used optical system (see stochastic localization microscopy techniques). [23] In Structured Illumination Light Sheet Microscopy, structured illumination techniques have been applied to further improve the optical sectioning capacity of light sheet fluorescence microscopy. [24]

Stripe artifacts

Top: stripe artifacts in light sheet fluorescence microscopy. Bottom: Reducing stripe artifacts by pivoting Lsfm stripes.svg
Top: stripe artifacts in light sheet fluorescence microscopy. Bottom: Reducing stripe artifacts by pivoting

As the illumination typically penetrates the sample from one side, obstacles lying in the way of the lightsheet can disturb its quality by scattering and/or absorbing the light. This typically leads to dark and bright stripes in the images. If parts of the samples have a significantly higher refractive index (e.g. lipid vesicles in cells), they can also lead to a focussing effect resulting in bright stripes behind these structures. To overcome this artifact, the lightsheets can e.g. be "pivoting". That means that the lightsheet's direction of incidence is changed rapidly (~1 kHz rate) by a few degrees (~10°), so light also hits the regions behind the obstacles. Illumination can also be performed with two (pivoted) lightsheets (see above) to further reduce these artifacts. [8] Alternatively, the Variational Stationary Noise Remover (VSNR) algorithm has been developed and is available as a free Fiji plugin. [35]

History

At the beginning of the 20th century, R. A. Zsigmondy introduced the ultramicroscope as a new illumination scheme into dark-field microscopy. Here sunlight or a white lamp is used to illuminate a precision slit. The slit is then imaged by a condensor lens into the sample to form a lightsheet. Scattering (sub-diffractive) particles can be observed perpendicularly with a microscope. This setup allowed the observation of particles with sizes smaller than the microscope's resolution and led to a Nobel prize for Zsigmondy in 1925. [36]

The first application of this illumination scheme for fluorescence microscopy was published in 1993 by Voie et al. under the name orthogonal-plane fluorescence optical sectioning (OPFOS). [4] for imaging of the internal structure of the cochlea. The resolution at that time was limited to 10 µm laterally and 26 µm longitudinally but at a sample size in the millimeter range. The orthogonal-plane fluorescence optical sectioning microscope used a simple cylindrical lens for illumination. Further development and improvement of the selective plane illumination microscope started in 2004. [5] After this publication by Huisken et al. the technique found wide application and is still adapted to new measurement situations today (see above). Since 2010 a first ultramicroscope with fluorescence excitation and limited resolution [37] and since 2012 a first selective plane illumination microscope are available commercially. [38] A good overview about the development of selective plane illumination microscopy is given in ref. [39] During 2012 also open source projects have started to appear that freely publish complete construction plans for light sheet fluorescence microscopes and also the required software suites. [40] [41] [42] [43]

Applications

Selective plane illumination microscopy/light sheet fluorescence microscopy is often used in developmental biology, where it enables long-time (several days) observations of embryonic development (even with full lineage tree reconstruction). [5] [44] Selective plane illumination microscopy can also be combined with techniques, like fluorescence correlation spectroscopy to allow spatially resolved mobility measurements of fluorescing particles (e.g. fluorescent beads, quantum dots or fluorescent proteins) inside living biological samples. [20] [21]

Strongly scattering biological tissue such as brain or kidney has to be chemically fixed and cleared before it can be imaged in a selective plane illumination microscope. [45] Special tissue clearing techniques have been developed for this purpose, e.g. 3DISCO, CUBIC and CLARITY. Depending on the index of refraction of the cleared sample, matching immersion fluids and special long-distance objectives must be used during imaging.

Related Research Articles

<span class="mw-page-title-main">Microscopy</span> Viewing of objects which are too small to be seen with the naked eye

Microscopy is the technical field of using microscopes to view objects and areas of objects that cannot be seen with the naked eye. There are three well-known branches of microscopy: optical, electron, and scanning probe microscopy, along with the emerging field of X-ray microscopy.

<span class="mw-page-title-main">Microscope</span> Scientific instrument

A microscope is a laboratory instrument used to examine objects that are too small to be seen by the naked eye. Microscopy is the science of investigating small objects and structures using a microscope. Microscopic means being invisible to the eye unless aided by a microscope.

<span class="mw-page-title-main">Optical microscope</span> Microscope that uses visible light

The optical microscope, also referred to as a light microscope, is a type of microscope that commonly uses visible light and a system of lenses to generate magnified images of small objects. Optical microscopes are the oldest design of microscope and were possibly invented in their present compound form in the 17th century. Basic optical microscopes can be very simple, although many complex designs aim to improve resolution and sample contrast.

A total internal reflection fluorescence microscope (TIRFM) is a type of microscope with which a thin region of a specimen, usually less than 200 nanometers can be observed.

<span class="mw-page-title-main">Fluorescence microscope</span> Optical microscope that uses fluorescence and phosphorescence

A fluorescence microscope is an optical microscope that uses fluorescence instead of, or in addition to, scattering, reflection, and attenuation or absorption, to study the properties of organic or inorganic substances. "Fluorescence microscope" refers to any microscope that uses fluorescence to generate an image, whether it is a simple set up like an epifluorescence microscope or a more complicated design such as a confocal microscope, which uses optical sectioning to get better resolution of the fluorescence image.

<span class="mw-page-title-main">Confocal microscopy</span> Optical imaging technique

Confocal microscopy, most frequently confocal laser scanning microscopy (CLSM) or laser scanning confocal microscopy (LSCM), is an optical imaging technique for increasing optical resolution and contrast of a micrograph by means of using a spatial pinhole to block out-of-focus light in image formation. Capturing multiple two-dimensional images at different depths in a sample enables the reconstruction of three-dimensional structures within an object. This technique is used extensively in the scientific and industrial communities and typical applications are in life sciences, semiconductor inspection and materials science.

<span class="mw-page-title-main">Two-photon excitation microscopy</span> Fluorescence imaging technique

Two-photon excitation microscopy is a fluorescence imaging technique that is particularly well-suited to image scattering living tissue of up to about one millimeter in thickness. Unlike traditional fluorescence microscopy, where the excitation wavelength is shorter than the emission wavelength, two-photon excitation requires simultaneous excitation by two photons with longer wavelength than the emitted light. The laser is focused onto a specific location in the tissue and scanned across the sample to sequentially produce the image. Due to the non-linearity of two-photon excitation, mainly fluorophores in the micrometer-sized focus of the laser beam are excited, which results in the spatial resolution of the image. This contrasts with confocal microscopy, where the spatial resolution is produced by the interaction of excitation focus and the confined detection with a pinhole.

A 4Pi microscope is a laser scanning fluorescence microscope with an improved axial resolution. With it the typical range of the axial resolution of 500–700 nm can be improved to 100–150 nm, which corresponds to an almost spherical focal spot with 5–7 times less volume than that of standard confocal microscopy.

<span class="mw-page-title-main">Near-field scanning optical microscope</span>

Near-field scanning optical microscopy (NSOM) or scanning near-field optical microscopy (SNOM) is a microscopy technique for nanostructure investigation that breaks the far field resolution limit by exploiting the properties of evanescent waves. In SNOM, the excitation laser light is focused through an aperture with a diameter smaller than the excitation wavelength, resulting in an evanescent field on the far side of the aperture. When the sample is scanned at a small distance below the aperture, the optical resolution of transmitted or reflected light is limited only by the diameter of the aperture. In particular, lateral resolution of 6 nm and vertical resolution of 2–5 nm have been demonstrated.

Winfried Denk is a German physicist. He built the first two-photon microscope while he was a graduate student in Watt W. Webb's lab at Cornell University, in 1989.

<span class="mw-page-title-main">Optical sectioning</span> Imaging of focal planes within a thick sample

Optical sectioning is the process by which a suitably designed microscope can produce clear images of focal planes deep within a thick sample. This is used to reduce the need for thin sectioning using instruments such as the microtome. Many different techniques for optical sectioning are used and several microscopy techniques are specifically designed to improve the quality of optical sectioning.

Super-resolution microscopy is a series of techniques in optical microscopy that allow such images to have resolutions higher than those imposed by the diffraction limit, which is due to the diffraction of light. Super-resolution imaging techniques rely on the near-field or on the far-field. Among techniques that rely on the latter are those that improve the resolution only modestly beyond the diffraction-limit, such as confocal microscopy with closed pinhole or aided by computational methods such as deconvolution or detector-based pixel reassignment, the 4Pi microscope, and structured-illumination microscopy technologies such as SIM and SMI.

Photo-activated localization microscopy and stochastic optical reconstruction microscopy (STORM) are widefield fluorescence microscopy imaging methods that allow obtaining images with a resolution beyond the diffraction limit. The methods were proposed in 2006 in the wake of a general emergence of optical super-resolution microscopy methods, and were featured as Methods of the Year for 2008 by the Nature Methods journal. The development of PALM as a targeted biophysical imaging method was largely prompted by the discovery of new species and the engineering of mutants of fluorescent proteins displaying a controllable photochromism, such as photo-activatible GFP. However, the concomitant development of STORM, sharing the same fundamental principle, originally made use of paired cyanine dyes. One molecule of the pair, when excited near its absorption maximum, serves to reactivate the other molecule to the fluorescent state.

Endomicroscopy is a technique for obtaining histology-like images from inside the human body in real-time, a process known as ‘optical biopsy’. It generally refers to fluorescence confocal microscopy, although multi-photon microscopy and optical coherence tomography have also been adapted for endoscopic use. Commercially available clinical and pre-clinical endomicroscopes can achieve a resolution on the order of a micrometre, have a field-of-view of several hundred μm, and are compatible with fluorophores which are excitable using 488 nm laser light. The main clinical applications are currently in imaging of the tumour margins of the brain and gastro-intestinal tract, particularly for the diagnosis and characterisation of Barrett’s Esophagus, pancreatic cysts and colorectal lesions. A number of pre-clinical and transnational applications have been developed for endomicroscopy as it enables researchers to perform live animal imaging. Major pre-clinical applications are in gastro-intestinal tract, toumour margin detection, uterine complications, ischaemia, live imaging of cartilage and tendon and organoid imaging.

Lattice light-sheet microscopy is a modified version of light sheet fluorescence microscopy that increases image acquisition speed while decreasing damage to cells caused by phototoxicity. This is achieved by using a structured light sheet to excite fluorescence in successive planes of a specimen, generating a time series of 3D images which can provide information about dynamic biological processes.

Wide-field multiphoton microscopy refers to an optical non-linear imaging technique tailored for ultrafast imaging in which a large area of the object is illuminated and imaged without the need for scanning. High intensities are required to induce non-linear optical processes such as two-photon fluorescence or second harmonic generation. In scanning multiphoton microscopes the high intensities are achieved by tightly focusing the light, and the image is obtained by beam scanning. In wide-field multiphoton microscopy the high intensities are best achieved using an optically amplified pulsed laser source to attain a large field of view (~100 μm). The image in this case is obtained as a single frame with a CCD without the need of scanning, making the technique particularly useful to visualize dynamic processes simultaneously across the object of interest. With wide-field multiphoton microscopy the frame rate can be increased up to a 1000-fold compared to multiphoton scanning microscopy. Wide-field multiphoton microscopes are not yet commercially available, but working prototypes exist in several optics laboratories.

Super-resolution dipole orientation mapping (SDOM) is a form of fluorescence polarization microscopy (FPM) that achieved super resolution through polarization demodulation. It was first described by Karl Zhanghao and others in 2016. Fluorescence polarization (FP) is related to the dipole orientation of chromophores, making fluorescence polarization microscopy possible to reveal structures and functions of tagged cellular organelles and biological macromolecules. In addition to fluorescence intensity, wavelength, and lifetime, the fourth dimension of fluorescence—polarization—can also provide intensity modulation without the restriction to specific fluorophores; its investigation in super-resolution microscopy is still in its infancy.

Super-resolution photoacoustic imaging is a set of techniques used to enhance spatial resolution in photoacoustic imaging. Specifically, these techniques primarily break the optical diffraction limit of the photoacoustic imaging system. It can be achieved in a variety of mechanisms, such as blind structured illumination, multi-speckle illumination, or photo-imprint photoacoustic microscopy in Figure 1.

Three-photon microscopy (3PEF) is a high-resolution fluorescence microscopy based on nonlinear excitation effect. Different from two-photon excitation microscopy, it uses three exciting photons. It typically uses 1300 nm or longer wavelength lasers to excite the fluorescent dyes with three simultaneously absorbed photons. The fluorescent dyes then emit one photon whose energy is three times the energy of each incident photon. Compared to two-photon microscopy, three-photon microscopy reduces the fluorescence away from the focal plane by , which is much faster than that of two-photon microscopy by . In addition, three-photon microscopy employs near-infrared light with less tissue scattering effect. This causes three-photon microscopy to have higher resolution than conventional microscopy.

Structured illumination light sheet microscopy (SI-LSM) is an optical imaging technique used for achieving volumetric imaging with high temporal and spatial resolution in all three dimensions. It combines the ability of light sheet microscopy to maintain spatial resolution throughout relatively thick samples with the higher axial and spatial resolution characteristic of structured illumination microscopy. SI-LSM can achieve lateral resolution below 100 nm in biological samples hundreds of micrometers thick.

References

  1. ↑ Fadero, T.C.; et al. (2018). "LITE microscopy: Tilted light-sheet excitation of model organisms offers high resolution and low photobleaching". The Journal of Cell Biology. 217 (5): 1869–1882. doi:10.1083/jcb.201710087. PMC   5940309 . PMID   29490939.
  2. ↑ Keller, Philipp J.; Stelzer, Ernst H. K. (2006). "Lichtscheiben-Mikroskopie in der molekularen Zellbiophysik" (PDF). Laborwelt. 7 (5): 18–21. Archived from the original (PDF) on 20 January 2013. Retrieved 23 April 2013.
  3. ↑ Tomer, Raju; Lovett-Barron, Matthew; Kauvar, Isaac; Andalman, Aaron; Burns, Vanessa M.; Sankaran, Sethuraman; Grosenick, Logan; Broxton, Michael; Yang, Samuel; Deisseroth, Karl (2015). "SPED Light Sheet Microscopy: Fast Mapping of Biological System Structure and Function". Cell. 163 (7): 1796–1806. doi:10.1016/j.cell.2015.11.061. ISSN   0092-8674. PMC   4775738 . PMID   26687363.
  4. 1 2 A. H. Voie; D. H. Burns; F. A. Spelman (June 1993). "Orthogonal-plane fluorescence optical sectioning: Three-dimensional imaging of macroscopic biological specimens". Journal of Microscopy. 170 (3): 229–236. doi:10.1111/j.1365-2818.1993.tb03346.x. ISSN   0022-2720. PMID   8371260. S2CID   2901024.
  5. 1 2 3 Huisken, J.; Swoger, J.; Del Bene, F.; Wittbrodt, J.; Stelzer, E. H. (2004). "Optical sectioning deep inside live embryos by selective plane illumination microscopy". Science . 305 (5686): 1007–1009. Bibcode:2004Sci...305.1007H. CiteSeerX   10.1.1.456.2250 . doi:10.1126/science.1100035. PMID   15310904. S2CID   3213175.
  6. ↑ Timo Mappes; Norbert Jahr; Andrea Csaki; Nadine Vogler; Juergen Popp; Wolfgang Fritzsche (5 November 2012). "The Invention of Immersion Ultramicroscopy in 1912-The Birth of Nanotechnology?". Angewandte Chemie International Edition. 51 (45): 11208–11212. doi:10.1002/anie.201204688. ISSN   1433-7851. PMID   23065955.
  7. 1 2 Greger, K; Swoger, J; Stelzer, EH (February 2007). "Basic building units and properties of a fluorescence single plane illumination microscope". Rev Sci Instrum. 78 (2): 023705–023705–7. Bibcode:2007RScI...78b3705G. doi: 10.1063/1.2428277 . PMID   17578115.
  8. 1 2 3 Huisken, Jan; Stainier, Didier Y. R. (2007). "Even fluorescence excitation by multidirectional selective plane illumination microscopy (mSPIM)". Optics Letters. 32 (17): 2608–10. Bibcode:2007OptL...32.2608H. doi:10.1364/OL.32.002608. ISSN   0146-9592. PMID   17767321. S2CID   15231468.(subscription required)
  9. ↑ Tomer, Raju; Khairy, Khaled; Amat, Fernando; Keller, Philipp J (3 June 2012). "Quantitative high-speed imaging of entire developing embryos with simultaneous multiview light-sheet microscopy". Nature Methods. 9 (7): 755–763. doi:10.1038/nmeth.2062. ISSN   1548-7091. PMID   22660741. S2CID   14191130.(subscription required)
  10. ↑ Krzic, Uros; Gunther, Stefan; Saunders, Timothy E; Streichan, Sebastian J; Hufnagel, Lars (3 June 2012). "Multiview light-sheet microscope for rapid in toto imaging". Nature Methods. 9 (7): 730–733. doi:10.1038/nmeth.2064. ISSN   1548-7091. PMID   22660739. S2CID   13388657.(subscription required)
  11. ↑ Keller, P. J.; Schmidt, A. D.; Wittbrodt, J.; Stelzer, E.H.K. (14 November 2008). "Reconstruction of Zebrafish Early Embryonic Development by Scanned Light Sheet Microscopy" (PDF). Science. 322 (5904): 1065–1069. Bibcode:2008Sci...322.1065K. doi:10.1126/science.1162493. ISSN   0036-8075. PMID   18845710. S2CID   7594561.
  12. ↑ Fahrbach, F. O.; Rohrbach, A. (November 2010). "A line scanned light-sheet microscope with phase shaped self-reconstructing beams". Optics Express. 18 (23): 24229–24244. Bibcode:2010OExpr..1824229F. doi: 10.1364/oe.18.024229 . PMID   21164769.
  13. 1 2 Planchon, T. A.; Gao, L.; Milkie, D. E.; Davidson, M. W.; Galbraith, J. A.; Galbraith, C. G.; Betzig, E. (2011). "Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination". Nature Methods. 8 (5): 417–423. doi:10.1038/nmeth.1586. PMC   3626440 . PMID   21378978.
  14. ↑ Vettenburg, Tom; Dalgarno, Heather I C; Nylk, Jonathan; Coll-Lladó, Clara; Ferrier, David E K; Čižmár, Tomáš; Gunn-Moore, Frank J; Dholakia, Kishan (2014). "Light-sheet microscopy using an Airy beam" (PDF). Nature Methods. 11 (5): 541–544. doi:10.1038/nmeth.2922. hdl: 10023/5521 . PMID   24705473. S2CID   205422713.
  15. ↑ Kafian, Hosein; Lalenejad, Meelad; Moradi-Mehr, Sahar; Akbari Birgani, Shiva; Abdollahpour, Daryoush (2020). "Light-Sheet Fluorescence Microscopy with Scanning Non-diffracting beams". Scientific Reports. 10 (1): 8501. Bibcode:2020NatSR..10.8501K. doi: 10.1038/s41598-020-63847-2 . PMC   7244762 . PMID   32444603.
  16. ↑ Nylk, Jonathan; McCluskey, Kaley; Preciado, Miguel A.; Mazilu, Michael; Yang, Zhengyi; Gunn-Moore, Frank J.; Aggarwal, Sanya; Tello, Javier A.; Ferrier, David E. K. (1 April 2018). "Light-sheet microscopy with attenuation-compensated propagation-invariant beams". Science Advances. 4 (4): eaar4817. arXiv: 1708.02612 . Bibcode:2018SciA....4.4817N. doi:10.1126/sciadv.aar4817. PMC   5938225 . PMID   29740614.
  17. ↑ Dunsby, C. (2008). "Optically sectioned imaging by oblique plane microscopy". Optics Express. 16 (25): 20306–16. Bibcode:2008OExpr..1620306D. doi:10.1364/OE.16.020306. hdl: 10044/1/53595 . ISSN   1094-4087. PMID   19065169.
  18. ↑ Zeno Lavagnino; Francesca Cella Zanacchi; Emiliano Ronzitti; Alberto Diaspro (2013). "Two-photon excitation selective plane illumination microscopy (2PE-SPIM) of highly scattering samples: characterization and application". Optics Express. 21 (5): 5998–6008. Bibcode:2013OExpr..21.5998L. doi: 10.1364/OE.21.005998 . ISSN   1094-4087. PMID   23482168.
  19. ↑ Wolf S, Supatto W, Debregeas G, Mahou P, Kruglik SG, Sintes J, Beaurepaire E, Candelier R (May 2015). "Whole-brain functional imaging with two-photon light-sheet microscopy". Correspondence. Nature Methods . 12 (5): 379–80. doi:10.1038/nmeth.3371. PMID   25924070. S2CID   19746295.
  20. 1 2 3 4 Capoulade, J.; Wachsmuth, M.; Hufnagel, L.; Knop, M. (2011). "Quantitative fluorescence imaging of protein diffusion and interaction in living cells". Nature Biotechnology. 29 (9): 835–839. doi:10.1038/nbt.1928. PMID   21822256. S2CID   10493584.
  21. 1 2 3 Wohland, T.; Shi, X.; Sankaran, J.; Stelzer, E. H. (May 2010). "Single plane illumination fluorescence correlation spectroscopy (SPIM-FCS) probes inhomogeneous three-dimensional environments". Optics Express. 18 (10): 10627–10641. Bibcode:2010OExpr..1810627W. doi: 10.1364/oe.18.010627 . PMID   20588915.
  22. ↑ Klaus Greger; Manuel J. Neetz; Emmanuel G. Reynaud; Ernst H.K. Stelzer (2011). "Three-dimensional Fluorescence Lifetime Imaging with a Single Plane Illumination Microscope provides an improved Signal to Noise Ratio". Optics Express. 19 (21): 20743–50. Bibcode:2011OExpr..1920743G. doi: 10.1364/OE.19.020743 . ISSN   1094-4087. PMID   21997084.
  23. 1 2 Francesca Cella Zanacchi; Zeno Lavagnino; Michela Perrone Donnorso; Alessio Del Bue; Laura Furia; Mario Faretta; Alberto Diaspro (9 October 2011). "Live-cell 3D super-resolution imaging in thick biological samples". Nature Methods. 8 (12): 1047–1049. doi:10.1038/nmeth.1744. ISSN   1548-7091. PMID   21983925. S2CID   205420075.
  24. 1 2 Jerome Mertz; Jinhyun Kim (2010). "Scanning light-sheet microscopy in the whole mouse brain with HiLo background rejection". Journal of Biomedical Optics. 15 (1): 016027–016027–7. Bibcode:2010JBO....15a6027M. doi: 10.1117/1.3324890 . ISSN   1083-3668. PMC   2917465 . PMID   20210471.
  25. ↑ Friedrich, Mike; Gan, Qiang; Ermolayev, Vladimir; Harms, Gregory S. (2011). "STED-SPIM: Stimulated Emission Depletion Improves Sheet Illumination Microscopy Resolution". Biophysical Journal. 100 (8): L43–5. Bibcode:2011BpJ...100L..43F. doi: 10.1016/j.bpj.2010.12.3748 . PMC   3077687 . PMID   21504720..
  26. ↑ Fadero TC, et al. 2017. "LITE microscopy: a technique for high numerical aperture, low photobleaching fluorescence imaging", bioRxiv, doi : 10.1101/181644.
  27. ↑ Jorand R, et al. 2012. "Deep and Clear Optical Imaging of Thick Inhomogeneous Samples", PLOS one, doi : 10.1371/journal.pone.0035795.
  28. ↑ Jorand R, 2013. "Amélioration des voies de détection et d'illumination d'un microscope SPIM pour l'imagerie 3D des sphéroïdes", theses.fr.
  29. ↑ Maizel, Alexis; von Wangenheim, Daniel; Federici, Fernán; Haseloff, Jim; Stelzer, Ernst H.K. (October 2011). "High-resolution live imaging of plant growth in near physiological bright conditions using light sheet fluorescence microscopy". The Plant Journal. 68 (2): 377–385. doi: 10.1111/j.1365-313X.2011.04692.x . ISSN   0960-7412. PMID   21711399.
  30. ↑ Terrence F. Holekamp; Diwakar Turaga; Timothy E. Holy (13 March 2008). "Fast Three-Dimensional Fluorescence Imaging of Activity in Neural Populations by Objective-Coupled Planar Illumination Microscopy". Neuron. 57 (5): 661–672. doi: 10.1016/j.neuron.2008.01.011 . ISSN   0896-6273. PMID   18341987. S2CID   9571663.
  31. ↑ Y. Wu; A. Ghitani; R. Christensen; A. Santella; Z. Du; G. Rondeau; Z. Bao; D. Colon-Ramos; H. Shroff (25 October 2011). "Inverted selective plane illumination microscopy (iSPIM) enables coupled cell identity lineaging and neurodevelopmental imaging in Caenorhabditis elegans". Proceedings of the National Academy of Sciences. 108 (43): 17708–17713. Bibcode:2011PNAS..10817708W. doi: 10.1073/pnas.1108494108 . ISSN   0027-8424. PMC   3203761 . PMID   22006307.
  32. 1 2 3 Wu, Yicong; Wawrzusin, Peter; Senseney, Justin; Fischer, Robert S; Christensen, Ryan; Santella, Anthony; York, Andrew G; Winter, Peter W; Waterman, Clare M; Bao, Zhirong; Colón-Ramos, Daniel A; McAuliffe, Matthew; Shroff, Hari (2013). "Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy". Nature Biotechnology. 31 (11): 1032–1038. doi:10.1038/nbt.2713. ISSN   1087-0156. PMC   4105320 . PMID   24108093.
  33. ↑ "Webpage Huisken Lab". Archived from the original on 7 July 2014.
  34. ↑ Jörg G. Ritter; Roman Veith; Jan-Peter Siebrasse; Ulrich Kubitscheck (2008), "High-contrast single-particle tracking by selective focal plane illumination microscopy", Optics Express, vol. 16, no. 10, pp. 7142–52, Bibcode:2008OExpr..16.7142R, doi: 10.1364/OE.16.007142 , ISSN   1094-4087, PMID   18545417
  35. ↑ Jerome Fehrenbach; Pierre Weiss; Corinne Lorenzo (2012). "Variational Algorithms to Remove Stationary Noise: Applications to Microscopy Imaging" (PDF). IEEE Transactions on Image Processing. 21 (10): 4420–4430. Bibcode:2012ITIP...21.4420F. doi:10.1109/TIP.2012.2206037. ISSN   1057-7149. PMID   22752131. S2CID   6828193.
  36. ↑ Nobel lecture of R. A. Zsigmondy: Properties of colloids (including a short explanation of the ultramicroscope)
  37. ↑ press release of LaVision Biotech Archived 24 December 2013 at the Wayback Machine (accessed 2012-11-04)
  38. ↑ Carl Zeiss press release about the Lightsheet Z.1 Light Sheet Microscope System (accessed 2012-11-15)
  39. ↑ P. A. Santi (1 February 2011). "Light Sheet Fluorescence Microscopy: A Review". Journal of Histochemistry & Cytochemistry. 59 (2): 129–138. doi:10.1369/0022155410394857. ISSN   0022-1554. PMC   3201139 . PMID   21339178.
  40. ↑ OpenSPIM project webpage (accessed 2013-06-08)
  41. ↑ Peter G Pitrone; Johannes Schindelin; Luke Stuyvenberg; Stephan Preibisch; Michael Weber; Kevin W Eliceiri; Jan Huisken; Pavel Tomancak (9 June 2013). "OpenSPIM: an open-access light-sheet microscopy platform". Nature Methods. 10 (7): 598–9. arXiv: 1302.1987 . doi:10.1038/nmeth.2507. ISSN   1548-7091. PMC   7450513 . PMID   23749304.
  42. ↑ The OpenSPIN project webpage (accessed 2013-06-08)
  43. ↑ Emilio J Gualda, Tiago Vale, Pedro Almada, Jos A Feij, Gabriel G Martins, Nuno Moreno (9 June 2013). "OpenSpinMicroscopy: an open-source integrated microscopy platform". Nature Methods. 10 (7): 599–600. doi:10.1038/nmeth.2508. ISSN   1548-7091. PMID   23749300. S2CID   27935584.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  44. ↑ Verveer, P. J.; Swoger, J.; Pampaloni, F.; Greger, K.; Marcello, M.; Stelzer, E. H. (2007). "High-resolution three-dimensional imaging of large specimens with light sheet-based microscopy". Nature Methods . 4 (4): 311–313. doi:10.1038/nmeth1017. PMID   17339847. S2CID   12440781.
  45. ↑ Jährling, Nina; Becker, Klaus; Wegenast-Braun, Bettina M.; Grathwohl, Stefan A.; Jucker, Mathias; Dodt, Hans-Ulrich (27 May 2015). Vitorica, Javier (ed.). "Cerebral β-Amyloidosis in Mice Investigated by Ultramicroscopy". PLOS ONE. 10 (5): e0125418. Bibcode:2015PLoSO..1025418J. doi: 10.1371/journal.pone.0125418 . ISSN   1932-6203. PMC   4446269 . PMID   26017149.
  46. ↑ Corinne Lorenzo; Céline Frongia; Raphael Jorand; Jérome Fehrenbach; Pierre Weiss; Amina Maandhui; Guillaume Gay; Bernard Ducommun; Valérie Lobjois (2011). "Live cell division dynamics monitoring in 3D large spheroid tumor models using light sheet microscopy". Cell Division. 6 (1): 22. doi: 10.1186/1747-1028-6-22 . ISSN   1747-1028. PMC   3274476 . PMID   22152157.
  47. ↑ Daisuke Takao; Atsushi Taniguchi; Takaaki Takeda; Seiji Sonobe; Shigenori Nonaka; Alexandre J. Kabla (5 December 2012), "High-Speed Imaging of Amoeboid Movements Using Light-Sheet Microscopy", PLOS ONE, vol. 7, no. 12, p. e50846, Bibcode:2012PLoSO...750846T, doi: 10.1371/journal.pone.0050846 , ISSN   1932-6203, PMC   3515486 , PMID   23227214

Further reading

This page is based on this Wikipedia article
Text is available under the CC BY-SA 4.0 license; additional terms may apply.
Images, videos and audio are available under their respective licenses.