Microfluidic trap array for massively parallel imaging of Drosophila embryos

Levario, Thomas J.; Zhan, Min; Lim, Bomyi; Shvartsman, Stanislav Y.; Lu, Hang

doi:10.1038/nprot.2013.034

Cited by 56 publications

(47 citation statements)

References 31 publications

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“…Recently, microfluidic devices and Micro-Electrical-Mechanical Systems (MEMS) have been successfully used in many Drosophila assays, [38][39][40][41][42][43][44][45][46] mainly because of their capability to provide scalable and controllable environments for quantitative embryonic [38][39][40][41][42][43] and larval [44][45][46] investigations in flies. These miniaturized devices that are made primarily from the elastomer polydimethylsiloxane (PDMS) have significantly enhanced the throughput, accuracy, and reproducibility of these assays.…”

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confidence: 99%

Agar-polydimethylsiloxane devices for quantitative investigation of oviposition behaviour of adult Drosophila melanogaster

et al. 2015

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Drosophila melanogaster (fruit fly) is a model organism and its behaviours including oviposition (egg-laying) on agar substrates have been widely used for assessment of a variety of biological processes in flies. Physical and chemical properties of the substrate are the dominant factors affecting Drosophila's oviposition, but they have not been investigated precisely and parametrically with the existing manual approaches. As a result, many behavioral questions about Drosophila oviposition, such as the combined effects of the aforementioned substrate properties (e.g., exposure area, sugar content, and stiffness) on oviposition and viability, and their threshold values, are yet to be answered. In this paper, we have devised a simple, easily implementable, and novel methodology that allows for modification of physical and chemical composition of agar substrates in order to quantitatively study survival and oviposition of adult fruit flies in an accurate and repeatable manner. Agar substrates have been modified by surface patterning using single and hexagonally arrayed through-hole polydimethylsiloxane (PDMS) membranes with various diameters and interspacing, as well as by substrate stiffness and sugar content modification via alteration of chemical components. While pure PDMS substrates showed a significant lethal effect on flies, a 0.5 mm diameter through-hole access to agar was found to abruptly increase the survival of adult flies to more than 93%. Flies avoided ovipositing on pure PDMS and on top of substrates with 0.5 mm diameter agar exposure areas. At a hole diameter of 2 mm (i.e., 0.25% exposure area) or larger, eggs were observed to be laid predominately inside the through-holes and along the edges of the PDMS-agar interface, showing a trending increase in site selection with 4 mm (i.e., 1% exposure area threshold) demonstrating natural oviposition rates similar to pure agar. The surfacemodified agar-PDMS hybrid devices and the threshold values reported for the substrate physical and chemical conditions affecting oviposition are novel; therefore, we advocate their use for future in-depth studies of oviposition behaviour in Drosophila melanogaster with accuracy and repeatability. The technique is also useful for development of novel assays for learning and decision-making studies as well as miniaturized devices for self-assembly of eggs and embryonic developmental investigations. V C 2015 AIP Publishing LLC. [http://dx

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confidence: 99%

Agar-polydimethylsiloxane devices for quantitative investigation of oviposition behaviour of adult Drosophila melanogaster

et al. 2015

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“…The traps are connected by a main channel for aggregate loading and perfusion of media and reagents. Loading of individual aggregates into traps is accomplished by a previously demonstrated hydrodynamic loading mechanism 49–51 that is dependent on the ratio of two flow paths in the device (Fig. 1e).…”

Section: Resultsmentioning

confidence: 99%

A microfluidic trap array for longitudinal monitoring and multi-modal phenotypic analysis of individual stem cell aggregates

Jackson-Holmes

McDevitt

2017

Lab Chip

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Three-dimensional pluripotent stem cell (PSC) cultures have the ability to undergo differentiation, self-organization, and morphogenesis to yield complex, in vitro tissue models that recapitulate key elements of native tissues. These tissue models offer a system for studying mechanisms of tissue development, investigating disease mechanisms, and performing drug screening. It remains challenging, however, to standardize PSC aggregate differentiation and morphogenesis methods due to heterogeneity stemming from biological and environmental sources. It is also difficult to monitor and assess large numbers of individual samples longitudinally throughout culture using typical batch-based culture methods. To address these challenges, we have developed a microfluidic platform for culture, longitudinal monitoring, and phenotypic analysis of individual stem cell aggregates. This platform uses a hydrodynamic loading principle to capture pre-formed stem cell aggregates in independent traps. We demonstrated that multi-day culture of aggregates in this platform reduces heterogeneity in phenotypic parameters such as size and morphology. Additionally, we showed that culture and analysis steps can be performed sequentially in the same platform, enabling correlation of multiple modes of analysis for individual samples. We anticipate this platform being applied to improve abilities for phenotypic analysis of PSC aggregate tissues and to facilitate research in standardizing culture systems in order to dually increase the yield and reduce the heterogeneity of PSC-derived tissues.

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“…Recently, Levario et al [47] presented a microfluidic trap array to rapidly orient hundreds of Drosophila embryos. In developmental studies, gradients of diffusible molecules are often used to induce morphogen concentration-dependent patterns.…”

Section: Organismsmentioning

confidence: 99%

High-throughput screening approaches and combinatorial development of biomaterials using microfluidics

2016

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Microfluidics, being a technology characterized by the engineered manipulation of fluids at the submillimeter scale, offers some interesting tools that can advance biomedical research and development. Screening platforms based on microfluidic technologies that allow high-throughput and combinatorial screening may lead to breakthrough discoveries not only in basic research but also relevant to clinical application. This is further strengthened by the fact that reliability of such screens may improve, since microfluidic systems allow close mimicking of physiological conditions. Finally, microfluidic systems are also very promising as micro factories of a new generation of natural or synthetic biomaterials and constructs, with finely controlled properties.

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Microfluidic trap array for massively parallel imaging of Drosophila embryos

Cited by 56 publications

References 31 publications

Agar-polydimethylsiloxane devices for quantitative investigation of oviposition behaviour of adult Drosophila melanogaster

Agar-polydimethylsiloxane devices for quantitative investigation of oviposition behaviour of adult Drosophila melanogaster

A microfluidic trap array for longitudinal monitoring and multi-modal phenotypic analysis of individual stem cell aggregates

High-throughput screening approaches and combinatorial development of biomaterials using microfluidics

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