Fully automated cellular-resolution vertebrate screening platform with parallel animal processing

Chang, Tsung-Yao; Pardo-Martín, Carlos; Allalou, Amin; Wählby, Carolina; Yanik, Mehmet Fatih

doi:10.1039/c1lc20849g

Cited by 103 publications

(81 citation statements)

References 12 publications

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“…If a higher imaging resolution is needed, than the setup is adapted in a way that the large particle flow cytometer technology can be used for pre-screening and subsequently analyze the samples at a medium throughput at a higher resolution. This can be done using the Vertebrate Automated Screening Technology 17,18 . This device can automatically collect live or fixed embryos between 2 and 7 days post fertilization from a multi well plate or bulk container, image 360° through a capillary using CLSM or stereo microscopy and dispose again in 2 bulk containers allowing manual sorting of the embryos based on the microscopic images.…”

Section: Discussionmentioning

confidence: 99%

Establishment and Optimization of a High Throughput Setup to Study Staphylococcus epidermidis and Mycobacterium marinum Infection as a Model for Drug Discovery

Veneman

Marín-Juez

Sonneville

et al. 2014

JoVE

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Zebrafish are becoming a valuable tool in the preclinical phase of drug discovery screenings as a whole animal model with high throughput screening possibilities. They can be used to bridge the gap between cell based assays at earlier stages and in vivo validation in mammalian models, reducing, in this way, the number of compounds passing through to testing on the much more expensive rodent models. In this light, in the present manuscript is described a new high throughput pipeline using zebrafish as in vivo model system for the study of Staphylococcus epidermidis and Mycobacterium marinum infection. This setup allows the generation and analysis of large number of synchronous embryos homogenously infected. Moreover the flexibility of the pipeline allows the user to easily implement other platforms to improve the resolution of the analysis when needed. The combination of the zebrafish together with innovative high throughput technologies opens the field of drug testing and discovery to new possibilities not only because of the strength of using a whole animal model but also because of the large number of transgenic lines available that can be used to decipher the mode of action of new compounds.

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Section: Discussionmentioning

confidence: 99%

Establishment and Optimization of a High Throughput Setup to Study Staphylococcus epidermidis and Mycobacterium marinum Infection as a Model for Drug Discovery

Veneman

Marín-Juez

Sonneville

et al. 2014

JoVE

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“…12 Furthermore, potential organ-specific drug toxicity problems can be rapidly identified using zebrafish transgenic models. [13][14] Compound library screens can also be performed with a remarkable throughput in contrats to conventional rodent models [15][16][17] Notwithstanding this, laboratory automation of in-situ analysis using zebrafish embryo and larvae is still deeply in its infancy. The major hurdle to widespread deployment of zebrafish embryo and larvae in large-scale drug development projects is lack of enabling high-throughput analytical platforms.…”

Section: Introductionmentioning

confidence: 99%

An integrated micromechanical large particle in flow sorter (MILPIS)

et al. 2015

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At present, the major hurdle to widespread deployment of zebrafish embryo and larvae in large-scale drug development projects is lack of enabling high-throughput analytical platforms. In order to spearhead drug discovery with the use of zebrafish as a model, platforms need to integrate automated pre-test sorting of organisms (to ensure quality control and standardization) and their in-test positioning (suitable for high-content imaging) with modules for flexible drug delivery. The major obstacle hampering sorting of millimetre sized particles such as zebrafish embryos on chip-based devices is their substantial diameter (above one millimetre), mass (above one milligram), which both lead to rapid gravitational-induced sedimentation and high inertial forces. Manual procedures associated with sorting hundreds of embryos are very monotonous and as such prone to significant analytical errors due to operator's fatigue. In this work, we present an innovative design of a micromechanical large particle in-flow sorter (MILPIS) capable of analysing, sorting and dispensing living zebrafish embryos for drug discovery applications. The system consisted of a microfluidic network, revolving micromechanical receptacle actuated by robotic servomotor and opto-electronic sensing module. The prototypes were fabricated in poly(methyl methacrylate) (PMMA) transparent thermoplastic using infrared laser micromachining. Elements of MILPIS were also fabricated in an optically transparent VisiJet resin using 3D stereolithography (SLA) processes (ProJet 7000HD, 3D Systems). The device operation was based on a rapidly revolving miniaturized mechanical receptacle. The latter function was to hold and position individual fish embryos for (i) interrogation, (ii) sorting decision-making and (iii) physical sorting..The system was designed to separate between fertilized (LIVE) and non-fertilized (DEAD) eggs, based on optical transparency using infrared (IR) emitters and receivers embedded in the system. Digital oscilloscope were used to distinguish the diffraction signals from IR sensors when both LIVE and DEAD embryos were flow through in the chip. Image process analysis were also used as detection module to track DEAD embryos as it flowed in the channel. ; phone 61 3 992 57157; fax 61 3 992 57110; http://www.rmit.edu.au/staff/donald-wlodkowic

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“…Relatively simple real-time image processing routines are applied to support, for example, automated sorting or selective image acquisition. Examples include the development of flowbased systems for high-throughput imaging and laser microsurgery of larvae ''on the fly,'' 59 workflows for the automatic injection of bacteria into the yolk, 22 image-based sorting robots, 33,61 a screening platform with automated manipulation of zebrafish including orienting and positioning regions of interest within the microscope's field of view, 25 and automated zooming into ROI for heartbeat detection. 67 …”

Section: Figmentioning

confidence: 99%

Automated Processing of Zebrafish Imaging Data: A Survey

et al. 2013

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Due to the relative transparency of its embryos and larvae, the zebrafish is an ideal model organism for bioimaging approaches in vertebrates. Novel microscope technologies allow the imaging of developmental processes in unprecedented detail, and they enable the use of complex image-based read-outs for highthroughput/high-content screening. Such applications can easily generate Terabytes of image data, the handling and analysis of which becomes a major bottleneck in extracting the targeted information. Here, we describe the current state of the art in computational image analysis in the zebrafish system. We discuss the challenges encountered when handling high-content image data, especially with regard to data quality, annotation, and storage. We survey methods for preprocessing image data for further analysis, and describe selected examples of automated image analysis, including the tracking of cells during embryogenesis, heartbeat detection, identification of dead embryos, recognition of tissues and anatomical landmarks, and quantification of behavioral patterns of adult fish. We review recent examples for applications using such methods, such as the comprehensive analysis of cell lineages during early development, the generation of a three-dimensional brain atlas of zebrafish larvae, and high-throughput drug screens based on movement patterns. Finally, we identify future challenges for the zebrafish image analysis community, notably those concerning the compatibility of algorithms and data formats for the assembly of modular analysis pipelines.

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Fully automated cellular-resolution vertebrate screening platform with parallel animal processing

Cited by 103 publications

References 12 publications

Establishment and Optimization of a High Throughput Setup to Study <em>Staphylococcus epidermidis</em> and <em>Mycobacterium marinum</em> Infection as a Model for Drug Discovery

Establishment and Optimization of a High Throughput Setup to Study <em>Staphylococcus epidermidis</em> and <em>Mycobacterium marinum</em> Infection as a Model for Drug Discovery

An integrated micromechanical large particle in flow sorter (MILPIS)

Automated Processing of Zebrafish Imaging Data: A Survey

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