Mitochondrial mass was quantified in PBMCs after stimulation +/− fatty acids using Mitotracker green fluorescence. PBMCs were incubated with αCD3/αCD28 ± 400 µM palmitate and/or oleate for 40 h then were washed twice with 1X PBS and incubated with 40 nM Mitotracker green (9074S, Cell Signaling Technology, Danvers, MA, USA) in 5 mM glucose-containing RPMI for 30 min. Mitotracker green fluorescence was assessed via fluorescence spectrophotometry (Biotek, Winooski, VT, USA) and the results were normalized to cell numbers.
Mitotracker green
Manufactured by Agilent Technologies
Sourced in United States
Mitotracker Green is a fluorescent dye used to label and track mitochondria in live cells. It passively diffuses across the cell membrane and accumulates in active mitochondria, allowing for visualization and monitoring of mitochondrial morphology and distribution.
Automatically generated - may contain errors
Lab products found in correlation
Mitotracker green, by Cell Signaling Technology (2 mentions)
Fluorescence spectrophotometry, by Agilent Technologies (1 mentions)
Mitotracker green, by Thermo Fisher Scientific (1 mentions)
Fluorescence microscope, by Thermo Fisher Scientific (1 mentions)
Confocal microscope, by Thermo Fisher Scientific (1 mentions)
Lionheart fx livingcell imaging analysis system, by Agilent Technologies (1 mentions)
2 protocols using mitotracker green
1
Quantifying Mitochondrial Mass in Stimulated PBMCs
2
Imaging Transplanted Mitochondria via Fluorescent Probes
Check if the same lab product or an alternative is used in the 5 most similar protocols
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For in vitro imaging,
MitoTracker Green and Red (Cell Signaling Technology, Danvers, MA,
USA) were used to incubate with isolated mitochondria and receptor
cells, respectively. The combination of Cy5 marked PEP–TPP
and MitoTracker Green-labeled mitochondria was imaged by a confocal
microscope (Thermo Fisher Scientific). To observe the dynamic internalization
of transplanted mitochondria, PEP(Cy5)–TPP–mitochondria
was added into the culture medium. Then the fluorescence intensity
of MitoTracker Green and Cy5 was detected by the Lionheart FX living
cell imaging analysis system (BioTek, Winooski, VT, USA).
For
in vivo imaging, to detect the retention of transplanted PEP(Cy5)–TPP–mitochondria
(GFP), mouse hearts were imaged by a fluorescence detection system
(IVIS Lumina XRMS, USA) after reperfusion for 3, 6, and 24 h. Besides,
the Cox4i1-GFP marked mitochondria of formalin-fixed heart tissue
sections were imaged with a fluorescence microscope (Thermo Fisher
Scientific).
MitoTracker Green and Red (Cell Signaling Technology, Danvers, MA,
USA) were used to incubate with isolated mitochondria and receptor
cells, respectively. The combination of Cy5 marked PEP–TPP
and MitoTracker Green-labeled mitochondria was imaged by a confocal
microscope (Thermo Fisher Scientific). To observe the dynamic internalization
of transplanted mitochondria, PEP(Cy5)–TPP–mitochondria
was added into the culture medium. Then the fluorescence intensity
of MitoTracker Green and Cy5 was detected by the Lionheart FX living
cell imaging analysis system (BioTek, Winooski, VT, USA).
For
in vivo imaging, to detect the retention of transplanted PEP(Cy5)–TPP–mitochondria
(GFP), mouse hearts were imaged by a fluorescence detection system
(IVIS Lumina XRMS, USA) after reperfusion for 3, 6, and 24 h. Besides,
the Cox4i1-GFP marked mitochondria of formalin-fixed heart tissue
sections were imaged with a fluorescence microscope (Thermo Fisher
Scientific).
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