TY - JOUR
T1 - BioPhotonics workstation: A versatile setup for simultaneous optical manipulation, heat stress, and intracellular pH measurements of a live yeast cell
AU - Aabo, Thomas
AU - Bañas, Andrew Rafael
AU - Glückstad, Jesper
AU - Siegumfeldt, Henrik
AU - Arneborg, Nils
N1 - Copyright 2011 American Institute of Physics. This article may be downloaded for personal use only. Any other use requires prior permission of the author and the American Institute of Physics. The following article appeared in Review of Scientific Instruments, Volume 82, Issue 8 and may be found at http://rsi.aip.org/resource/1/rsinak/v82/i8/p083707_s1.
PY - 2011
Y1 - 2011
N2 - 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.
AB - 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.
U2 - 10.1063/1.3625274
DO - 10.1063/1.3625274
M3 - Journal article
SN - 0034-6748
VL - 82
JO - Review of Scientific Instruments
JF - Review of Scientific Instruments
IS - 8
ER -