Protocol detail

Mitochondrial ROS Quantification

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To assess levels in reactive oxygen species (ROS), cells were incubated with the mitochondrial oxygen free radical indicator MitoSOX-red (Life Technologies, Carlsbad, CA) for 30 min at 37 °C. Slides were then mounted for confocal imaging in an open perfusion microincubator (PDMI-2; Harvard Apparatus) and images were obtained at 561 nm excitation by using a confocal microscope (810; Carl Zeiss, Oberkochen, Germany). Optical densitometry quantifications were expressed as fluorescence intensity normalized to areas as previously described^{74 (link)}.

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Duggan M.R., Mohseni Ahooyi T., Parikh V, & Khalili K. (2021). Neuromodulation of BAG co-chaperones by HIV-1 viral proteins and H2O2: implications for HIV-associated neurological disorders. Cell Death Discovery, 7, 60.

Publication 2021

MitoSOX-red PDMI-2

Cells Confocal microscope Densitometry Fluorescence Free radical Mitochondrial Optical Oxygen Reactive oxygen species

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Variable analysis

independent variables

Incubation time with MitoSOX-red (30 min)

dependent variables

Levels of reactive oxygen species (ROS)

control variables

Incubation temperature (37 °C)
Imaging technique (confocal microscopy at 561 nm excitation)
Optical densitometry quantification method (fluorescence intensity normalized to areas)

controls

No explicit mention of positive or negative controls

Annotations

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