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

    Research output: Contribution to journalJournal articleResearchpeer-review

    6 Citations (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.

    Original languageEnglish
    JournalReview of Scientific Instruments
    Volume82
    Issue number8
    Number of pages6
    ISSN0034-6748
    DOIs
    Publication statusPublished - 2011

    Cite this