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

Thomas Ask Aabo; Andrew Raphael Banás; Jesper Glückstad; Henrik Siegumfeldt; Nils Arneborg

doi:10.1063/1.3625274

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

Thomas Ask Aabo, Andrew Raphael Banás, Jesper Glückstad, Henrik Siegumfeldt, Nils Arneborg

6 Citationer (Scopus)

Abstract

In this study we have modified the BioPhotonics workstation (BWS), which allows for using long working distance objective for optical trapping, to include traditional epi-fluorescence microscopy, using the trapping objectives. We have also added temperature regulation of sample stage, allowing for fast temperature variations while trapping. Using this modified BWS setup, we investigated the internal pH (pHi) response and membrane integrity of an optically trapped Saccharomyces cerevisiae cell at 5 mW subject to increasing temperatures. The pHi of the cell is obtained from the emission of 5-(and-6)-carboxyfluorescein diacetate, succinimidyl ester, at 435 and 485 nm wavelengths, while the permeability is indicated by the fluorescence of propidium iodide. We present images mapping the pHi and permeability of the cell at different temperatures and with enough spatial resolution to localize these attributes within the cell. The combined capability of optical trapping, fluorescence microscopy and temperature regulation offers a versatile tool for biological research.

Originalsprog	Engelsk
Tidsskrift	Review of Scientific Instruments
Vol/bind	82
Udgave nummer	8
Antal sider	6
ISSN	0034-6748
DOI	https://doi.org/10.1063/1.3625274
Status	Udgivet - aug. 2011

Adgang til dokumentet

10.1063/1.3625274

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Citationsformater

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abstract = "In this study we have modified the BioPhotonics workstation (BWS), which allows for using long working distance objective for optical trapping, to include traditional epi-fluorescence microscopy, using the trapping objectives. We have also added temperature regulation of sample stage, allowing for fast temperature variations while trapping. Using this modified BWS setup, we investigated the internal pH (pHi) response and membrane integrity of an optically trapped Saccharomyces cerevisiae cell at 5 mW subject to increasing temperatures. The pHi of the cell is obtained from the emission of 5-(and-6)-carboxyfluorescein diacetate, succinimidyl ester, at 435 and 485 nm wavelengths, while the permeability is indicated by the fluorescence of propidium iodide. We present images mapping the pHi and permeability of the cell at different temperatures and with enough spatial resolution to localize these attributes within the cell. The combined capability of optical trapping, fluorescence microscopy and temperature regulation offers a versatile tool for biological research.",

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AU - Aabo, Thomas Ask

AU - Banás, Andrew Raphael

AU - Glückstad, Jesper

AU - Siegumfeldt, Henrik

AU - Arneborg, Nils

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