Technical Information
Colour: Red/near-infrared
Detection method : Fluorescence, one & two-photon excitation
Peak excitation/emission wavelengths: Ex:542nm/Em:690nm
Two-photon absorption peak: 790nm
Two-photon cross-section: 224 GM
Form : Liquid (resuspended in water)
Sub-cellular localization: mitochondria
For use with: epifluorescence and confocal microscopes, flow cytometers
Fixable: no
Kit contents:
1 vial contains 125 µL of an aqueous solution with a concentration of 1 mM for a final volume of 250 mL of imaging medium
Storage: Store at 4°C for up to 7 months. For long-term storage, aliquot and store at -20°C.
Applications:
Experimental outputs: live imaging & flow cytometry
GlowMito produces a bright red labeling of the entire mitochondrial network, including those with reduced potential. It is primarily intended to visualize mitochondria based on their presence rather than assessing their electrochemical properties. GlowMito does not need to be washed out: mitochondrial fluorescence will remain stable for up to 3 days without causing toxicity and can therefore be integrated in your long-term experiments.
GlowMito is perfectly suitable for studying mitochondrial morphology, movement (velocity, localization), tracking, dynamics (fusion/fission), biogenesis (replication & growth), autophagy, etc.
GlowMito can be used in combination with other probes (GFP, potentiometric dyes, calcium signaling probes, etc).
We do not recommend its use for the measure of mitochondrial mass or volume density.
Tested and validated on:
- Human cells: HEK293, HeLa, MCF-7, MDA-MB-231, HMLE, UACC-62, U2-OS, Gli36, HAEC, SH-SY5Y, A-172, A549, patient-derived skeletal muscle cells & primary smooth muscle cells
- Mice cells: primary cortical neurons
- Monkey cells: COS-7
- Tissues: hiPSC-derived heart tissues & mice isolated pressurized blood vessels
- Parasitic protists: Trichomonas Vaginalis (hydrogenosome labeling)
GlowMito showed no internalization in yeast cells, Entamoeba histolytica, Giardia intestinalis parasites and seedlings.
Additional information:
Results
Smooth muscle cell mitochondria labeling in an intact pressurized cerebral artery using GlowMito
This confocal microscope image depicts the smooth muscle cell layer of an intact pressurized cerebral artery, illustrating mitochondria (shown in magenta) along with the nuclei (shown in blue). Smooth muscle cell nuclei are elongated and run perpendicular to the vessel axis. Small, rounded nuclei correspond to adventitial cells on the vessel wall.
Credits: Image captured by Safa in Dr. Charles Norton's laboratory - University of Missouri (US)
GlowMito produces a stable & bright labeling of mitochondria for 4 hours
GlowMito was directly added at 500 nM in live COS-7 culture medium. Cells were imaged repeatedly after 1h, 2.5h & 4h of incubation with GlowMito. Scale bar = 20µm.
Internalization & stability of GlowMito compared to a commercial mitochondrial probe
HEK293 cells were observed before and after 2, 4, and 6 minutes of incubation with GlowMito 500 nM.
Acquisition parameters: laser 2%, gain 15
A maximum of internalization is observed after 4 minutes.
Credits: Morgane LeMao, Guy Lenaers, Olivier Siri - MitoLab, Unité MitoVasc, Université d’Angers, CNRS UMR6015, Inserm U1083, Université de Marseille, CINAM UMR 7325 – 2023
