Analysis of mitochondrial DNA in microfluidic systems

Taylor, Patricia M.; Manage, Dammika P.; Helmle, Karmon; Zheng, Yao; Glerum, D. Moira; Backhouse, C.

doi:10.1016/j.jchromb.2005.05.025

Cited by 11 publications

(8 citation statements)

References 18 publications

(20 reference statements)

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“…To explore these two possibilities, we grew hippocampal neurons in microfluidic chambers [22] that enable independent manipulation of the fluid environment surrounding cell bodies and axons, as illustrated in Figure 5. Before addition of hydrogen peroxide, there was extensive mitochondrial transport in both the cell body and axonal compartments.…”

Section: Resultsmentioning

confidence: 99%

Oxidative stress inhibits axonal transport: implications for neurodegenerative diseases

Fang

Bourdette

Banker

2012

Mol Neurodegeneration

111

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BackgroundReactive oxygen species (ROS) released by microglia and other inflammatory cells can cause axonal degeneration. A reduction in axonal transport has also been implicated as a cause of axonal dystrophies and neurodegeneration, but there is a paucity of experimental data concerning the effects of ROS on axonal transport. We used live cell imaging to examine the effects of hydrogen peroxide on the axonal transport of mitochondria and Golgi-derived vesicles in cultured rat hippocampal neurons.ResultsHydrogen peroxide rapidly inhibited axonal transport, hours before any detectable changes in mitochondrial morphology or signs of axonal degeneration. Mitochondrial transport was affected earlier and was more severely inhibited than the transport of Golgi-derived vesicles. Anterograde vesicle transport was more susceptible to peroxide inhibition than retrograde transport. Axonal transport partially recovered following removal of hydrogen peroxide and local application of hydrogen peroxide inhibited transport, suggesting that the effects were not simply a result of nerve cell death. Sodium azide, an ATP synthesis blocker, had similar effects on axonal transport, suggesting that ATP depletion may contribute to the transport inhibition due to hydrogen peroxide.ConclusionsThese results indicate that inhibition of axonal transport is an early consequence of exposure to ROS and may contribute to subsequent axonal degeneration.

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Section: Resultsmentioning

confidence: 99%

Oxidative stress inhibits axonal transport: implications for neurodegenerative diseases

Fang

Bourdette

Banker

2012

Mol Neurodegeneration

111

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“…Figure 7 illustrates two types of CE chips and their operations. The CE chips are extensively used in medical research such as DNA analysis [ 44 ], infectious disease diagnostics [ 45 ] and sample purification [ 46 ]. Now some companies such as Micronit Microfluidics BV in The Netherlands, microLIQUID in USA, and Micralyne Inc. in Canada supply CE chips and kits for clinical diagnostics.…”

Section: Applications Of Opto-microfluidic Sensorsmentioning

confidence: 99%

Microfabrication and Applications of Opto-Microfluidic Sensors

Zhang

Men

Chen

2011

Sensors

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A review of research activities on opto-microfluidic sensors carried out by the research groups in Canada is presented. After a brief introduction of this exciting research field, detailed discussion is focused on different techniques for the fabrication of opto-microfluidic sensors, and various applications of these devices for bioanalysis, chemical detection, and optical measurement. Our current research on femtosecond laser microfabrication of optofluidic devices is introduced and some experimental results are elaborated. The research on opto-microfluidics provides highly sensitive opto-microfluidic sensors for practical applications with significant advantages of portability, efficiency, sensitivity, versatility, and low cost.

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“…Results indicated that the digestions were completed within 1-10 min and subsequent separations in the agarose-filled channels (4 cm effective length) were completed in 5-10 min (20 min total analysis time) [48]. Recently, an on-chip treatment of mtDNA employing restriction enzymes was used by Taylor et al [49] to differentiate homogeneous and heterogeneous populations of mtDNAs. The procedure was carried out on a straight channel 8.0 cm commercial glass chip (Micralyne) and included excising a mtDNA sequence from plasmid DNA via enzymatic digestion with EcoRI to linearize the plasmid loops into dsDNA and denaturing/renaturing to form duplexes of the digested DNA all taking place in a single sample well.…”

Section: Rflpmentioning

confidence: 99%

“…The generated products were labeled with an intercalating dye (SYTOX Orange) that was present in the separation matrix (Genescan polymer containing 10% glycerol) during electrophoretic separation (7.6 cm effective separation channel length). According to the authors, the method could be performed in about 45 min, whereas conventional methods would require days to perform this same type of assay [49].…”

Section: Rflpmentioning

confidence: 99%

High resolution DNA separations using microchip electrophoresis

Sinville

Soper

2007

J of Separation Science

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Planar microfluidic devices have emerged as effective tools for the electrophoretic separation of a variety of different DNA inputs. The advancement of this miniaturized platform was inspired initially by demands placed on electrophoretic performance metrics by the human genome project and has provided a viable alternative to slab gel and even capillary formats due to its ability to offer high resolution separations of nucleic acid materials in a fraction of the time associated with its predecessors, consumption of substantially less sample and reagents while maintaining the ability to perform many separations in parallel for realizing ultra-high throughputs. Another compelling advantage of this separation platform is that it offers the potential for integrating front-end sample preprocessing steps onto the separation device eliminating the need for manual sample handling. This review aims to compile a recent survey of various electrophoretic separations using either glass or polymer-based microchips in the areas of genotyping and DNA sequencing as well as those involving the growing field of DNA-based forensics.

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Analysis of mitochondrial DNA in microfluidic systems

Cited by 11 publications

References 18 publications

Oxidative stress inhibits axonal transport: implications for neurodegenerative diseases

Oxidative stress inhibits axonal transport: implications for neurodegenerative diseases

Microfabrication and Applications of Opto-Microfluidic Sensors

High resolution DNA separations using microchip electrophoresis

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