LITE microscopy: Tilted light-sheet excitation of model organisms offers high resolution and low photobleaching

Fadero, Tanner; Gerbich, Therese M.; Rana, K; Suzuki, Aussie; DiSalvo, Matthew; Schaefer, Kristina N.; Heppert, Jennifer K.; Boothby, Thomas E.; Goldstein, Bob; Peifer, Mark; Allbritton, Nancy L.; Gladfelter, Amy S.; Maddox, Amy Shaub; Maddox, Paul S.

doi:10.1083/jcb.201710087

Cited by 73 publications

(50 citation statements)

References 18 publications

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“…Studies that used H. exemplaris strain Z151 prior to the 2018 species disambiguation from H. dujardini (Gasiorek et al 2018) generally referred to the species as H. dujardini, or as Hypsibius cf. dujardini, and in many of these studies the strain was indicated as Z151 and/or sourced from Sciento Bavan et al 2009;Mali et al 2010;Cesari et al 2012;Beltrán-Pardo et al 2013;Horikawa et al 2013;Tenlen et al 2013;Smith and Jockusch 2014;Boothby et al 2015Boothby et al , 2017Gross and Mayer 2015;Kondo et al 2015;Arakawa et al 2016;Bemm et al 2016;Fernandez et al 2016;Hering et al 2016;Hyra et al 2016a,b;Kosztyła et al 2016;Koutsovoulos et al 2016;Levin et al 2016;Smith et al 2016Stec et al 2016;Erdmann et al 2017;Gross et al 2017Gross et al , 2018Vasanthan et al 2017;Yoshida et al 2017Yoshida et al , 2018Fadero et al 2018;Nelson 2018;Rost-Roszkowska et al 2018). One study using Thulinia stephaniae (Hejnol and Schnabel 2005) indicates some contrasting developmental features in a second tardigrade species.…”

Section: Related Speciesmentioning

confidence: 99%

The Emergence of the TardigradeHypsibius exemplarisas a Model System

Goldstein

2018

Cold Spring Harb Protoc

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The success of scientists in revealing biological mechanisms has depended in large part on choosing tractable model systems. In 1997, molecular phylogenetics revealed that two of biology's most tractable models-Caenorhabditis elegans and Drosophila-are much more closely related to each other than had been thought previously. I began to explore whether any of the little-studied members of this branch of the tree of life might serve as a new model for comparative biology that could make use of the rich and ongoing sources of information flowing from C. elegans and Drosophila research. Tardigrades, also known as water bears, make up a phylum of microscopic animals. The tardigrade Hypsibius exemplaris (recently disambiguated from a closely related species, Hypsibius dujardini) can be maintained in laboratories and has a generation time of <2 wk at room temperature. Stocks of animals can be stored frozen and revived. The animals and their embryos are optically clear, and embryos are laid in groups, with each synchronous clutch of embryos laid in a clear molt. We have developed techniques for laboratory study of this system, including methods for microinjection of animals, immunolocalization, in situ hybridization, RNA interference, transcriptomics, and methods for identifying proteins that mediate tolerance to extreme environments. Here, I review the development of this animal as an emerging model system, as well as recent molecular studies aimed at understanding the evolution of developmental mechanisms that underpin the evolution of animal form and at understanding how biological materials can survive extreme environments.

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Section: Related Speciesmentioning

confidence: 99%

The Emergence of the TardigradeHypsibius exemplarisas a Model System

Goldstein

2018

Cold Spring Harb Protoc

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“…SPIM has been used to visualize large living biological systems such as zebrafish, 11,12 Drosophila melanogaster embryos, 13 Caenorhabditis elegans, 14 tumor cell spheroids, 15,16 and Arabidopsis thaliana. 17 Different LSFM configurations have been proposed to perform multicolor 3-D imaging. [18][19][20][21] For instance, Krieger et al performed dual color fluorescence imaging using a single camera and two separate color channels, while Jahr et al used a diffractive unit to spectrally split the images onto a camera in order to obtain hyperspectral images.…”

Section: Introductionmentioning

confidence: 99%

Multicolor fluorescence microscopy using static light sheets and a single-channel detection

et al. 2019

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We present a multicolor fluorescence microscope system, under a selective plane illumination microscopy (SPIM) configuration, using three continuous wave-lasers and a single-channel-detection camera. The laser intensities are modulated with three time-delayed pulse trains that operate synchronously at one third of the camera frame rate, allowing a sequential excitation and an image acquisition of up to three different biomarkers. The feasibility of this imaging acquisition mode is demonstrated by acquiring single-plane multicolor images of living hyphae of Neurospora crassa. This allows visualizing simultaneously the localization and dynamics of different cellular components involved in apical growth in living hyphae. The configuration presented represents a noncommercial, cost-effective alternative microscopy system for the rapid and simultaneous acquisition of multifluorescent images and can be potentially useful for three-dimensional imaging of large biological samples.

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“…The most notable of these techniques is lattice light sheet, where a two-dimensional optical lattice is used to create thin, structured illumination sheets that are dithered during a single exposure [14]. Traditionally LSFM requires two objective lenses, one for illumination and one for detection, placed at a 90-degree angle relative to each other such that the light sheet from the illumination source coincides with the imaging plane of the detection objective [14][15][16][17][18][19][20][21]. Such systems can be less user-friendly than conventional microscopes.…”

Section: Introductionmentioning

confidence: 99%

VIEW-MOD: A Versatile Illumination Engine With a Modular Optical Design for Fluorescence Microscopy

Liu

Hobson

Pimenta

et al. 2019

Preprint

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We developed VIEW-MOD (Versatile Illumination Engine With a Modular Optical Design): a compact, multi-modality microscope, which accommodates multiple illumination schemes including variable angle total internal reflection, point scanning and vertical/horizontal light sheet. This system allows combining and flexibly switching between different illuminations and imaging modes by employing three electrically tunable lenses and two faststeering mirrors. This versatile optics design provides control of 6 degrees of freedom of the illumination source (3 translation, 2 tilt, and beam shape) plus the axial position of the imaging plane. We also developed standalone software with an easy-to-use GUI to calibrate and control the microscope. We demonstrate the applications of this system and software in biosensor imaging, optogenetics and fast 3D volume imaging. This system is ready to fit into complex imaging circumstances requiring precise control of illumination and detection paths, and has a broad scope of usability for a myriad of biological applications. the detection pathway. Through using some or all of the modules available, our system can implement a variety of imaging modalities. Here, we outline the theory and purpose of each module, with the reader directed to Figure 1 for the optical layout.

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LITE microscopy: Tilted light-sheet excitation of model organisms offers high resolution and low photobleaching

Cited by 73 publications

References 18 publications

The Emergence of the TardigradeHypsibius exemplarisas a Model System

The Emergence of the TardigradeHypsibius exemplarisas a Model System

Multicolor fluorescence microscopy using static light sheets and a single-channel detection

VIEW-MOD: A Versatile Illumination Engine With a Modular Optical Design for Fluorescence Microscopy

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