Continuous-flow C. elegans fluorescence expression analysis with real-time image processing through microfluidics

Caenorhabditis elegans ( C. elegans ) has been a popular model organism for several decades since its first discovery of the huge research potential for modeling human diseases and genetics. Sorting is an important means of providing stage- or age-synchronized worm populations for many worm-based bioassays. However, conventional manual techniques for C. elegans sorting are tedious and inefficient, and commercial complex object parametric analyzer and sorter is too expensive and bulky for most laboratories. Recently, the development of lab-on-a-chip (microfluidics) technology has greatly facilitated C. elegans studies where large numbers of synchronized worm populations are required and advances of new designs, mechanisms, and automation algorithms. Most previous reviews have focused on the development of microfluidic devices but lacked the summaries and discussion of the biological research demands of C. elegans , and are hard to read for worm researchers. We aim to comprehensively review the up-to-date microfluidic-assisted C. elegans sorting developments from several angles to suit different background researchers, i.e., biologists and engineers. First, we highlighted the microfluidic C. elegans sorting devices' advantages and limitations compared to the conventional commercialized worm sorting tools. Second, to benefit the engineers, we reviewed the current devices from the perspectives of active or passive sorting, sorting strategies, target populations, and sorting criteria. Third, to benefit the biologists, we reviewed the contributions of sorting to biological research. We expect, by providing this comprehensive review, that each researcher from this multidisciplinary community can effectively find the needed information and, in turn, facilitate future research.

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Section: Microfluidic Sorting Of C Elegans and Dev...mentioning

confidence: 99%

Microfluidic-Assisted Caenorhabditis elegans Sorting: Current Status and Future Prospects

Yuan

Duan

et al. 2023

Cyborg Bionic Syst

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“…The above devices rely on physical or thermal contact with animals in order to immobilize them. Recently, Yan et al [ 90 ] developed an approach in which individual worms with different levels of GFP expression were encapsulated in oil droplets and analyzed fluorescently in an automated setup. The system provided 100% accuracy and an analysis speed of 0.5 s per worm, although resolution was poor.…”

Section: Microfluidic Devices To Perform Fast Automated and Highmentioning

confidence: 99%

Microfluidic Approaches for Manipulating, Imaging, and Screening C. elegans

Gupta

Rezai

2016

Micromachines

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The nematode C. elegans (worm) is a small invertebrate animal widely used in studies related to fundamental biological processes, disease modelling, and drug discovery. Due to their small size and transparent body, these worms are highly suitable for experimental manipulations. In recent years several microfluidic devices and platforms have been developed to accelerate worm handling, phenotypic studies and screens. Here we review major tools and briefly discuss their usage in C. elegans research.

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“…As PDMS microfluidic chambers can be produced via imprinting, chamber size and dimensions can be customized to suit specific experimental conditions, allowing precise control over the culture environment. Unlike in vivo models, an in vitro PDMS microfluidic platform allows acquisition of cellular responses at a single cell level 22,26–30 . The malleability of PDMS microfluidic chambers not only allows micro-scaled volumes, which reduces the amount of culture medium required, but also facilitates the design of various models to simulate biophysical mechanics, such as shear stress, tension, and compression 22,26,29,31,32 .…”

Section: Introductionmentioning

confidence: 99%

An in vitro pressure model towards studying the response of primary retinal ganglion cells to elevated hydrostatic pressures

Mak

Chan

et al. 2019

Sci Rep

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Glaucoma is a leading cause of blindness characterized by progressive degeneration of retinal ganglion cells (RGCs). A well-established risk factor for the development and progression of glaucoma is elevation of intraocular pressure (IOP). However, how elevated IOP leads to RGC degeneration remains poorly understood. Here, we fabricate a facile, tunable hydrostatic pressure platform to study the effect of increased hydrostatic pressure on RGC axon and total neurite length, cell body area, dendritic branching, and cell survival. The hydrostatic pressure can be adjusted by varying the height of a liquid reservoir attached to a three-dimensional (3D)-printed adapter. The proposed platform enables long-term monitoring of primary RGCs in response to various pressure levels. Our results showed pressure-dependent changes in the axon length, and the total neurite length. The proportion of RGCs with neurite extensions significantly decreased by an average of 38 ± 2% (mean ± SEM) at pressures 30 mmHg and above (p < 0.05). The axon length and total neurite length decreased at a rate of 1.65 ± 0.18 μm and 4.07 ± 0.34 μm, respectively (p < 0.001), for each mmHg increase in pressure after 72 hours pressure treatment. Dendritic branching increased by 0.20 ± 0.05 intersections/day at pressures below 25 mmHg, and decreased by 0.07 ± 0.01 intersections/day at pressures above 25 mmHg (p < 0.001). There were no significant changes in cell body area under different levels of hydrostatic pressure (p ≥ 0.05). Application of this model will facilitate studies on the biophysical mechanisms that contribute to the pathophysiology of glaucoma and provide a channel for the screening of potential pharmacological agents for neuroprotection.

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Continuous-flow C. elegans fluorescence expression analysis with real-time image processing through microfluidics

Cited by 18 publications

References 44 publications

Microfluidic-Assisted Caenorhabditis elegans Sorting: Current Status and Future Prospects

Microfluidic-Assisted Caenorhabditis elegans Sorting: Current Status and Future Prospects

Microfluidic Approaches for Manipulating, Imaging, and Screening C. elegans

An in vitro pressure model towards studying the response of primary retinal ganglion cells to elevated hydrostatic pressures

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