Continuous live cell imaging using dark field microscopy

Zeng, Yang; Rui, Cao; Zhu, Jie; Zhao, Wei; Zhang, Ce

doi:10.1039/d2ay00043a

Cited by 2 publications

(2 citation statements)

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“…The droplet shape can then be adjusted by tuning the hydrophobicity of the substrate (i.e., the contact angle) via polyvinyl alcohol processing following plasma treatment (Trantidou et al, 2017 ). To prevent unwanted evaporation, the droplet fluidic device was positioned in a sealed box at 100% RH, and a temperature of 28.5°C to maintain a healthy environment for zebrafish (Zeng et al, 2022 ).…”

Section: Resultsmentioning

confidence: 99%

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A fluidic platform for mobility evaluation of zebrafish with gene deficiency

Jia

Feng

et al. 2023

Front. Mol. Neurosci.

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IntroductionZebrafish is a suitable animal model for molecular genetic tests and drug discovery due to its characteristics including optical transparency, genetic manipulability, genetic similarity to humans, and cost-effectiveness. Mobility of the zebrafish reflects pathological conditions leading to brain disorders, disrupted motor functions, and sensitivity to environmental challenges. However, it remains technologically challenging to quantitively assess zebrafish's mobility in a flowing environment and simultaneously monitor cellular behavior in vivo.MethodsWe herein developed a facile fluidic device using mechanical vibration to controllably generate various flow patterns in a droplet housing single zebrafish, which mimics its dynamically flowing habitats.ResultsWe observe that in the four recirculating flow patterns, there are two equilibrium stagnation positions for zebrafish constrained in the droplet, i.e., the “source” with the outward flow and the “sink” with the inward flow. Wild-type zebrafish, whose mobility remains intact, tend to swim against the flow and fight to stay at the source point. A slight deviation from streamline leads to an increased torque pushing the zebrafish further away, whereas zebrafish with motor neuron dysfunction caused by lipin-1 deficiency are forced to stay in the “sink,” where both their head and tail align with the flow direction. Deviation angle from the source point can, therefore, be used to quantify the mobility of zebrafish under flowing environmental conditions. Moreover, in a droplet of comparable size, single zebrafish can be effectively restrained for high-resolution imaging.ConclusionUsing the proposed methodology, zebrafish mobility reflecting pathological symptoms can be quantitively investigated and directly linked to cellular behavior in vivo.

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Section: Resultsmentioning

confidence: 99%

“…Repeated knocking of the metal spring on the glass slide causes vibration, which propagates across the slides and reaches the droplet. Consequently, mechanically caused deformation of the droplet leads to variations in the internal flow patterns (Zeng et al, 2022 ).…”

Section: Methodsmentioning

confidence: 99%