BioPhotonics workstation: A versatile setup for simultaneous optical manipulation, heat stress, and intracellular pH measurements of a live yeast cell

Aabo, Thomas; Bañas, Andrew Rafael; Glückstad, Jesper; Siegumfeldt, Henrik; Arneborg, Nils

doi:10.1063/1.3625274

Cited by 9 publications

(9 citation statements)

References 22 publications

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“…( pH int ). This result was expected, as the temperature, because of the experimental set-up, did not change within that time period [16]. However, after the first 2 min, s.d.…”

Section: Resultsmentioning

confidence: 69%

“…Once a stable temperature is reached, the chamber is stable within +0.28C. For more details, see Aabo et al [16].…”

Section: Heating Stagementioning

confidence: 99%

“…A non-standard fluorescence microscope was used to acquire images [16]. The excitation source was a monochromator ( polychrome II) from Till photonics.…”

Section: Fluorescence Ratio Imaging Microscopymentioning

confidence: 99%

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Intracellular pH distribution as a cell health indicator in Saccharomyces cerevisiae

Aabo

Glückstad

Siegumfeldt

et al. 2011

J. R. Soc. Interface.

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Internal pH regulation is vital for many cell functions, including transport mechanisms and metabolic enzyme activity. More specifically, transport mechanisms are to a wide degree governed by internal pH distributions. We introduce the term standard deviation of the intracellular pH (s.d.( pH int )) to describe the internal pH distributions. The cellular pH distributional response to external stress such as heat has not previously been determined. In this study, the intracellular pH ( pH i ) and the s.d.( pH int ) of Saccharomyces cerevisiae cells exposed to supralethal temperatures were measured using fluorescence ratio imaging microscopy (FRIM). An exponential decline in pH i was observed after an initial small decline. For the first time, we report the use of FRIM for determining in vivo plasma membrane proton permeability coefficients in yeast. Furthermore, the exponential decay of pH i and the rupture of the cell plasma membrane, as measured by propidium iodide staining, at 708C were not simultaneous but were separated by a significant temporal difference. Finally, a nonlinear relationship between the pH i and s.d.( pH int ) was found; i.e. the s.d.( pH int ) was significantly more sensitive to supralethal temperatures than pH i . s.d.( pH int ) is therefore proposed as an early health/vitality indicator in S. cerevisiae cells exposed to heat stress.

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“…( pH int ). This result was expected, as the temperature, because of the experimental set-up, did not change within that time period [16]. However, after the first 2 min, s.d.…”

Section: Resultsmentioning

confidence: 69%

“…Once a stable temperature is reached, the chamber is stable within +0.28C. For more details, see Aabo et al [16].…”

Section: Heating Stagementioning

confidence: 99%

See 1 more Smart Citation

Intracellular pH distribution as a cell health indicator in Saccharomyces cerevisiae

Aabo

Glückstad

Siegumfeldt

et al. 2011

J. R. Soc. Interface.

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“…3.8(b)). This allows room for a variety of auxiliary applications such as advanced micro-spectroscopic [26] or multi-photon characterization methods or pH mapping of heat stressed cells through fluorescence ratio imaging [70]. This has also allowed direct side view imaging, making three dimensional experiments more intuitive ( Fig.…”

Section: Tradeoffs Of the Bwsmentioning

confidence: 99%

A Next Generation BioPhotonics Workstation

Glückstad

Palima

Tauro

et al. 2011

Optics in the Life Sciences

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, whose contributions remain relevant to our present work. The BRC group that hosted my external stay and fabricated the microtools that we trap, Lόránd Kelemen, Pál Ormos, Gaszton Vizsnyiczai, Badri Aekbote, András Búzás. Collaborators at Aarhus University and Copenhagen University as well as the people at the business development side for helping me appreciate the things I build in the laboratory. Rune Christiansen and the rest of DTU Danchip for fabricating our matched filters. And the Danish Research Council (FTP) for funding. I would also like to express my thanks to my examiners, Tomáš Čižmár and Simon Hanna whose comments have helped improved this thesis. Kirstine Berg-Sørensen, also for the helpful comments and for facilitating practical matters regarding the thesis. For the less technical aspects that nonetheless contribute to the grand scheme of things, as well as my mental health, I also thank my family for understanding my extended absence, my officemates and neighbours for standing in as co-PhDs, my very kind landlady, Birgit, for not minding all those scattered computers, cables and musical instruments, and the very nice or strange people I meet everyday at work.

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“…We tested the idea of optically steerable freestanding waveguides using our BioPhotonics Workstation (BWS). 4 BWS uses real-time reconfigurable counter-propagating beam traps controlled by direct spatial mapping from an addressable light-shaping module. Axial manipulation is achieved by balancing the intensity ratios of the counterpropagating beams.…”

mentioning

confidence: 99%