Simultaneous monitoring of oxygen consumption and acidification rates of a single zebrafish embryo during embryonic development within a microfluidic device

Huang, Shih-Hao; Huang, Kuo-Sheng; Liou, Yan-Min

doi:10.1007/s10404-016-1841-z

Cited by 12 publications

(18 citation statements)

References 25 publications

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“…Figure 4 shows a schematic of the optical detection system, equipped with a UV LED as the light modulation source, to determine the OCR of a single C. elegans in real time via phase-based phosphorescence lifetime detection. The details of the facility setup and data acquisition were described in our previous work [ 23 , 24 , 25 ]. Briefly, the optical detection system utilized a 3 W high power UV LED (390 nm, Edison Opto Corp., Taipei, Taiwan) as the light modulation source.…”

Section: Methodsmentioning

confidence: 99%

Bioenergetic Health Assessment of a Single Caenorhabditis elegans from Postembryonic Development to Aging Stages via Monitoring Changes in the Oxygen Consumption Rate within a Microfluidic Device

Huang

Lin

2018

Sensors

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Monitoring dynamic changes in oxygen consumption rates (OCR) of a living organism in real time provide an indirect method of monitoring changes in mitochondrial function during development, aging, or malfunctioning processes. In this study, we developed a microfluidic device integrated with an optical detection system to measure the OCR of a single developing Caenorhabditis elegans (C. elegans) from postembryonic development to aging stages in real time via phase-based phosphorescence lifetime measurement. The device consists of two components: an acrylic microwell deposited with an oxygen-sensitive luminescent layer for oxygen (O2) measurement and a microfluidic module with a pneumatically driven acrylic lid to controllably seal the microwell. We successfully measured the basal respiration (basal OCR, in pmol O2/min/worm) of a single C. elegans inside a microwell from the stages of postembryonic development (larval stages) through adulthood to aged adult. Sequentially adding metabolic inhibitors to block bioenergetic pathways allowed us to measure the metabolic profiles of a single C. elegans at key growth and aging stages, determining the following fundamental parameters: basal OCR, adenosine triphosphate (ATP)-linked OCR, maximal OCR, reserve respiratory capacity, OCR due to proton leak, and non-mitochondrial OCR. The bioenergetic health index (BHI) was calculated from these fundamental parameters to assess the bioenergetic health of a single developing C. elegans from the postembryonic development to aging stages. The changes in BHI are correlated to C. elegans development stage, with the highest BHI = 27.5 for 4-day-old adults, which possess well-developed bioenergetic functionality. Our proposed platform demonstrates for the first time the feasibility of assessing the BHI of a single C. elegans from postembryonic development to aging stages inside a microfluidic device and provides the potential for a wide variety of biomedical applications that relate mitochondrial malfunction and diseases.

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Section: Methodsmentioning

confidence: 99%

Bioenergetic Health Assessment of a Single Caenorhabditis elegans from Postembryonic Development to Aging Stages via Monitoring Changes in the Oxygen Consumption Rate within a Microfluidic Device

Huang

Lin

2018

Sensors

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“…To select well‐developed embryos, researchers have developed embryo characterization and evaluation platforms. The oxygen consumption, the utilization rate of glucose and other morphological parameters can be measured during in vitro embryo culture . To assess the viability of embryos, Wu et al designed a chip with a built‐in amperometric detector (Figure A), which can measure the oxygen consumption of a single embryo in situ.…”

Section: Bioengineered Microenvironment For Early Embryo Culture In Vmentioning

confidence: 99%

“…Huang et al prepared a microfluidic device with a dual‐luminous sensor (Figure D), which can simultaneously monitor changes in oxygen consumption and pH values. The device is easy to manufacture and can facilitate high‐throughput monitoring …”

Section: Bioengineered Microenvironment For Early Embryo Culture In Vmentioning

confidence: 99%

“…Copyright 2008, American Chemical Society (C) Three sensors in the microchamber were designed to measure the concentrations of dissolved oxygen, glucose and lactic acid in embryo metabolism . Copyright 2019, Elsevier BV (D) A light modulated microfluidic device was developed for multiple long‐term measurements of oxygen consumption and acid extrusion in embryo metabolism . Copyright 2016, Springer‐Verlag Berlin Heidelberg…”

Section: Bioengineered Microenvironment For Early Embryo Culture In Vmentioning

confidence: 99%

“…Copyright 2019, Elsevier BV (D) A light modulated microfluidic device was developed for multiple long‐term measurements of oxygen consumption and acid extrusion in embryo metabolism . Copyright 2016, Springer‐Verlag Berlin Heidelberg…”

Section: Bioengineered Microenvironment For Early Embryo Culture In Vmentioning

confidence: 99%

See 2 more Smart Citations

Bioengineered microenvironment to culture early embryos

Guo

Wang

et al. 2020

Cell Proliferation

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The abnormalities of early post‐implantation embryos can lead to early pregnancy loss and many other syndromes. However, it is hard to study embryos after implantation due to the limited accessibility. The success of embryo culture in vitro can avoid the challenges of embryonic development in vivo and provide a powerful research platform for research in developmental biology. The biophysical and chemical cues of the microenvironments impart significant spatiotemporal effects on embryonic development. Here, we summarize the main strategies which enable researchers to grow embryos outside of the body while overcoming the implantation barrier, highlight the roles of engineered microenvironments in regulating early embryonic development, and finally discuss the future challenges and new insights of early embryo culture.

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Soft lithography fabrication of index-matched microfluidic devices for reducing artifacts in fluorescence and quantitative phase imaging

Kim

Kim²,

Kim

et al. 2017

Microfluid Nanofluid

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Microfluidic devices are widely used for biomedical applications based on microscopy or other optical detection methods. However, the materials commonly used for microfabrication typically have a high refractive index relative to water, which can create artifacts at device edges and limit applicability to applications requiring high precision imaging or morphological feature detection. Here we present a soft lithography method to fabricate microfluidic devices out of MY133-V2000, a UV-curable, fluorinated polymer with low refractive index that is close to that of water (n = 1.33). The primary challenge in the use of this material (and fluorinated materials in general) is the low adhesion of the fluorinated material; we present several alternative fabrication methods we have tested to improve inter-layer adhesion. The close match between the refractive index of this material and aqueous solutions commonly used in biomedical applications enables fluorescence imaging at microchannel or other microfabricated edges without distortion. The close match in refractive index also enables quantitative phase microscopy (QPM) imaging across the full width of microchannels without error-inducing artifacts for measurement of cell biomass. Overall, our results demonstrate the utility of low-refractive index microfluidics for biological applications requiring high precision optical imaging.

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Simultaneous monitoring of oxygen consumption and acidification rates of a single zebrafish embryo during embryonic development within a microfluidic device

Cited by 12 publications

References 25 publications

Bioenergetic Health Assessment of a Single Caenorhabditis elegans from Postembryonic Development to Aging Stages via Monitoring Changes in the Oxygen Consumption Rate within a Microfluidic Device

Bioenergetic Health Assessment of a Single Caenorhabditis elegans from Postembryonic Development to Aging Stages via Monitoring Changes in the Oxygen Consumption Rate within a Microfluidic Device

Bioengineered microenvironment to culture early embryos

Soft lithography fabrication of index-matched microfluidic devices for reducing artifacts in fluorescence and quantitative phase imaging

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