Mitochondrial Membrane Potential Imaging

HeLa cells were plated in six-well plates containing 0.2% gelatin–coated glass coverslips (Thapa et al., 2011 (link)), and 100 nM TMRE perchlorate was added to cells and incubated for 30 min at 37°C. Nuclear DNA stain, 2-(4-ethoxyphenyl)-6-[6-(4-methylpiperazin-1-yl)-1H-benzimidazol-2-yl]-1H-benzimidazole (Hoescht 33342), was added for 5 min. Image acquisition was performed using a Carl Zeiss 510 confocal microscope using a 63× oil objective with excitation at 561 and 405 nm, respectively. Images were quantified using ImageJ (Madesh et al., 2005 (link)).

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Hoffman N.E., Chandramoorthy H.C., Shanmughapriya S., Zhang X.Q., Vallem S., Doonan P.J., Malliankaraman K., Guo S., Rajan S., Elrod J.W., Koch W.J., Cheung J.Y, & Madesh M. (2014). SLC25A23 augments mitochondrial Ca2+ uptake, interacts with MCU, and induces oxidative stress–mediated cell death. Molecular Biology of the Cell, 25(6), 936-947.

Publication 2014

510 confocal microscope

Benzimidazole Cells Confocal microscope Gelatin Hela cells Perchlorate Stain

Corresponding Organization :

Other organizations : Temple University

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Variable analysis

independent variables

TMRE perchlorate concentration (100 nM)
Incubation time (30 min)

dependent variables

Mitochondrial membrane potential (measured by TMRE fluorescence)

control variables

Cell type (HeLa cells)
Cell culture vessel (six-well plates)
Coverslip coating (0.2% gelatin)
Staining (Hoescht 33342 for nuclear DNA)
Imaging system (Carl Zeiss 510 confocal microscope, 63× oil objective)
Excitation wavelengths (561 nm and 405 nm)

controls

No positive or negative controls were explicitly mentioned in the provided information.

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