A difference imaging technique for monitoring real-time changes in morphology within the cell, tissue, and organism spatial domain

McLamore, Eric S.; Stensberg, Matthew; Yale, Gowri; Ochoa‐Acuña, Hugo; Sepúlveda, Marı́a S.; Sun, Xiaoming; Akkuş, Ozan; Porterfield, Diane

doi:10.1117/12.851694

Cited by 4 publications

(4 citation statements)

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“…Root elongation rate (RER) was measured in 4‐day maize roots before and after external addition of 15 μ m IAA using an image subtraction technique (McLamore et al. , 2010a).…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…DVIT software was used to control video imaging hardware and image acquisition. Pixel changes were recorded at a 1 kHz acquisition rate in the visible spectra, and image subtraction was conducted against a reference image according to McLamore et al (2010a). Processed images were displayed against a grey background, and no convolution kernels or image alteration were used during post-processing.…”

Section: Root Growth Rate Measurementsmentioning

confidence: 99%

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Non-invasive quantification of endogenous root auxin transport using an integrated flux microsensor technique

et al. 2010

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SUMMARYIndole-3-acetic acid (IAA) is a primary phytohormone that regulates multiple aspects of plant development. Because polar transport of IAA is an essential determinant of organogenesis and dynamic tropic growth, methods to monitor IAA movement in vivo are in demand. A self-referencing electrochemical microsensor was optimized to non-invasively measure endogenous IAA flux near the surface of Zea mays roots without the addition of exogenous IAA. Enhanced sensor surface modification, decoupling of acquired signals, and integrated flux analyses were combined to provide direct, real time quantification of endogenous IAA movement in B73 maize inbred and brachytic2 (br2) auxin transport mutant roots. BR2 is localized in epidermal and hypodermal tissues at the root apex. br2 roots exhibit reduced shootward IAA transport at the root apex in radiotracer experiments and reduced gravitropic growth. IAA flux data indicates that maximal transport occurs in the distal elongation zone of maize roots, and net transport in/out of br2 roots was decreased compared to B73. Integration of short term real time flux data in this zone revealed oscillatory patterns, with B73 exhibiting shorter oscillatory periods and greater amplitude than br2. IAA efflux and influx were inhibited using 1-N-naphthylphthalamic acid (NPA), and 2-naphthoxyacetic acid (NOA), respectively. A simple harmonic oscillation model of these data produced a correlation between modeled and measured values of 0.70 for B73 and 0.69 for br2. These results indicate that this technique is useful for real-time IAA transport monitoring in surface tissues and that this approach can be performed simultaneously with current live imaging techniques.

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“…Root elongation rate (RER) was measured in 4‐day maize roots before and after external addition of 15 μ m IAA using an image subtraction technique (McLamore et al. , 2010a).…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Root Growth Rate Measurementsmentioning

confidence: 99%

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Non-invasive quantification of endogenous root auxin transport using an integrated flux microsensor technique

et al. 2010

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“…Root growth rate was measured using a high-resolution optical difference imaging technique (McLamore et al 2010b). High-resolution imaging was conducted using a Pulnix progressive scan camera/zoomscope (Sunnyvale, CA) on the stage incorporated with the optrode (see supplemental Figure S1).…”

Section: Phytoplankton Mat Oxygen Fluxmentioning

confidence: 99%

Self-referencing optrodes for measuring spatially resolved, real-time metabolic oxygen flux in plant systems

McLamore

Jaroch

Chatni

et al. 2010

Planta

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The ability to non-invasively measure metabolic oxygen flux is a very important tool for physiologists interested in a variety of questions ranging from basic metabolism, growth/development, and stress adaptation. Technologies for measuring oxygen concentration near the surface of cells/tissues include electrochemical and optical techniques. A wealth of knowledge was gained using these tools for quantifying real-time physiology. Fiber-optic microprobes (optrodes) have recently been developed for measuring oxygen in a variety of biomedical and environmental applications. We have adopted the use of these optical microsensors for plant physiology applications, and used the microsensors in an advanced sensing modality known as self-referencing. Self-referencing is a non-invasive microsensor technique used for measuring real-time flux of analytes. This paper demonstrates the use of optical microsensors for non-invasively measuring rhizosphere oxygen flux associated with respiration in plant roots, as well as boundary layer oxygen flux in phytoplankton mats. Highly sensitive/selective optrodes had little to no hysteresis/calibration drift during experimentation, and an extremely high signal-to-noise ratio. We have used this new tool to compare various aspects of rhizosphere oxygen flux for roots of Glycine max, Zea mays, and Phaseolus vulgaris, and also mapped developmentally relevant profiles and distinct temporal patterns. We also characterized real-time respiratory patterns during inhibition of cytochrome and alternative oxidase pathways via pharmacology. Boundary layer oxygen flux was also measured for a phytoplankton mat during dark:light cycling and exposure to pharamacological inhibitors. This highly sensitive technology enables non-invasive study of oxygen transport in plant systems under physiologically relevant conditions.

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Shear‐induced detachment of biofilms from hollow fiber silicone membranes

Huang

McLamore

Chuang

et al. 2012

Biotech & Bioengineering

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A suite of techniques was utilized to evaluate the correlation between biofilm physiology, fluid-induced shear stress, and detachment in hollow fiber membrane aerated bioreactors. Two monoculture species biofilms were grown on silicone fibers in a hollow fiber membrane aerated bioreactors (HfMBR) to assess detachment under laminar fluid flow conditions. Both physiology (biofilm thickness and roughness) and nutrient mass transport data indicated the presence of a steady state mature biofilm after 3 weeks of development. Surface shear stress proved to be an important parameter for predicting passive detachment for the two biofilms. The average shear stress at the surface of Nitrosomonas europaea biofilms (54.5 ± 3.2 mPa) was approximately 20% higher than for Pseudomonas aeruginosa biofilms (45.8 ± 7.7 mPa), resulting in higher biomass detachment. No significant difference in shear stress was measured between immature and mature biofilms of the same species. There was a significant difference in detached biomass for immature vs. mature biofilms in both species. However, there was no difference in detachment rate between the two species.

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A difference imaging technique for monitoring real-time changes in morphology within the cell, tissue, and organism spatial domain

Cited by 4 publications

References 0 publications

Non-invasive quantification of endogenous root auxin transport using an integrated flux microsensor technique

Non-invasive quantification of endogenous root auxin transport using an integrated flux microsensor technique

Self-referencing optrodes for measuring spatially resolved, real-time metabolic oxygen flux in plant systems

Shear‐induced detachment of biofilms from hollow fiber silicone membranes

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