3D print customized sample holders for live light sheet microscopy

Jeandupeux, Emeric; Lobjois, Valérie; Ducommun, Bernard

doi:10.1016/j.bbrc.2015.06.072

Cited by 18 publications

(13 citation statements)

References 9 publications

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“…Confocal or light-sheet imaging in combination with innovative sample preparation methods (45,76) and staining protocols (77) can provide the necessary images. Evaluating these images with single-cell-based segmentation and a subsequent analysis with neighborhood graphs (17) can provide the necessary data to refine the complex models.…”

Section: Outlook: Experiments Inspired By the Mathematical Modelsmentioning

confidence: 99%

An Introduction to Image-Based Systems Biology of Multicellular Spheroids for Experimentalists and Theoreticians

Fischer

2019

Computational Biology

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Multicellular organisms are inherently three-dimensional. This leads to complex intercellular interactions that cannot be reproduced in two-dimensional cell culture. Instead, three-dimensional spheroids, ball-shaped cell aggregates, arise as model systems. Spheroids provide an accurate in vitro representation of the three-dimensional organization of cells in tissues, and compared to a real tissue, they excel with well-defined experimental conditions, easy handling, and suitability for high-quality imaging. Therefore, spheroids are an experimental system that can be readily combined with mathematical modeling. This chapter shows how image-based systems biology is implemented for multicellular spheroids to study three-dimensional cell-cell interactions. The chapter is intended for experimentalists and theoreticians who plan to extend their research by linkage with other disciplines. The relevant concepts for experimental approaches and quantitative imaging are introduced and linked to mathematical models of spheroids. This results in a list of potential systems biology workflows for typical spheroid research areas in cell biology, cancer biology, and bioprinting. In all three areas, there is a large gap between the details of the mathematical models and the

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Section: Outlook: Experiments Inspired By the Mathematical Modelsmentioning

confidence: 99%

An Introduction to Image-Based Systems Biology of Multicellular Spheroids for Experimentalists and Theoreticians

Fischer

2019

Computational Biology

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“…Reproduced from Ref. ( ) with permission from Elsevier. B : Illustrations of the PDMS microfluidic device for spheroid formation, culture, and imaging by SPIM.…”

Section: Parallelized Approaches To Increase Lsfm Throughputmentioning

confidence: 99%

Three‐dimensional imaging flow cytometry through light‐sheet fluorescence microscopy

et al. 2017

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Flow cytometry is the tool of choice for high-speed acquisition and analysis of large cell populations, with the tradeoff of lacking intracellular spatial information. Although in the last decades flow cytometry systems that can actually acquire two-dimensional spatial information were developed, some of the limitations remained though, namely constrains related to sample size and lack of depth or dynamic information. The combination of fluidics and light-sheet illumination has the potential to address these limitations. By having cells travelling with the flowing sheath one can, in a controlled fashion, force them at constant speed through the light-sheet enabling the synchronized acquisition of several optical sections, that is, three-dimensional imaging. This approach has already been used for imaging cellular spheroids, plankton, and zebra-fish embryos. In this review, we discuss the known solutions and standing challenges of performing three-dimensional high-throughput imaging of multicellular biological models using fluidics, while retaining cell and organelle-level resolution. © 2017 International Society for Advancement of Cytometry.

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“…Plant samples can be grown either directly into holders, such as glass capillaries or fluorinated ethylene propylene (FEP) tubes (Maizel et al, 2011;Sena et al, 2011;Kaufmann et al, 2012;Ovečka et al, 2015), or on a solid medium and then later introduced to a holder such a glass capillary or customized holding device (Fig. 1D) (Maizel et al, 2011;Kaufmann et al, 2012;Jeandupeux et al, 2015;Ovečka et al, 2015). An example of this can be seen in the imaging of the initial steps of plant germination, which is performed by positioning seeds inside jellified medium within a glass or FEP tube (Fig.…”

Section: What Are the Specific Challenges Of Light Sheet Microscopy?mentioning

confidence: 99%

Light sheet microscopy and live imaging of plants

Berthet

Maizel

2016

Journal of Microscopy

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SummaryLight sheet fluorescence microscopy (LSFM) is increasingly used to investigate biological processes in animals as well as in plants. LSFM achieves optical sectioning by the selective illumination of a single plane of the sample with a sheet of laser light while simultaneously recording emitted fluorescence orthogonally to the illumination plane. A 3D image of the sample can then be generated with a temporal resolution ranging from seconds to several days, and at scales ranging from subcellular to whole organ. By design, LSFM provides fast imaging, and low phototoxicity, two key criteria for live imaging under physiological conditions. Despite its potential, LSFM remains underutilized in plant biology. This review aims to highlight challenges of live imaging in plants, to describe key steps in using LSFM on live plant samples and finally at providing an overview of published examples of applications of LSFM in plants.

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3D print customized sample holders for live light sheet microscopy

Cited by 18 publications

References 9 publications

An Introduction to Image-Based Systems Biology of Multicellular Spheroids for Experimentalists and Theoreticians

An Introduction to Image-Based Systems Biology of Multicellular Spheroids for Experimentalists and Theoreticians

Three‐dimensional imaging flow cytometry through light‐sheet fluorescence microscopy

Light sheet microscopy and live imaging of plants

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