Overview
GlowMito is a cell permeant fluorescent probe that is selective for the mitochondria. Derived from a compound with high intrinsic fluorescence, GlowMito quickly penetrates cells and produces a bright labeling of mitochondria without inducing mitotoxicity or altering cell physiology. It has been successfully used for mitochondria imaging in live cells and tissues.
GlowMito is a fantastic illustration of the fruitful discoveries that can arise when organic chemistry meets biology, and it paves the way to a new class of fluorescent probes.
A technology designed by
Olivier Siri
and
Guy Lenaers
.
Images
Smooth muscle cell mitochondria labeling in an intact pressurized cerebral artery using GlowMito
Time-lapse epifluorescence microscopy images of COS-7 cells after 1h, 2.5h and 4h of incubation with GlowMito 500 nM. Scale bar = 20µm
Internalization of GlowMito (500 nM) compared to a commercial mitochondrial probe (500 nM)
Observation before and after 2, 4, and 6 minutes of HEK293 cells incubation.
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
HEK293 cells labeled with 500nM GlowMito – Scale bar=20µm
Credits: Morgane LeMao, Guy Lenaers, Olivier Siri - MitoLab, Unité MitoVasc, Université d’Angers, CNRS UMR6015, Inserm U1083, Université de Marseille, CINAM UMR 7325 – 2023
MCF7 cells labeled with 500nM GlowMito – Scale bar=20µm
Credits: Morgane LeMao, Guy Lenaers, Olivier Siri - MitoLab, Unité MitoVasc, Université d’Angers, CNRS UMR6015, Inserm U1083, Université de Marseille, CINAM UMR 7325 – 2023
UACC-62 cells labeled with 500nM GlowMito – Scale bar=20µm
redits: Morgane LeMao, Guy Lenaers, Olivier Siri MitoLab, Unité MitoVasc, Université d’Angers, CNRS UMR6015, Inserm U1083, Université de Marseille, CINAM UMR 7325 – 2023
COS-7 cells labeled with 500 nM GlowMito - Scale bar = 20µm
HeLa cells labeled with 500 nM GlowMito - Scale bar = 20µmFeatures
Based on non-covalent interactions and diluted in water, GlowMito is not toxic for biological samples and can be safely left in imaging medium for several hours. GlowMito also does not affect normal mitochondrial functions.
Ready-to-use
GlowMito is provided as a stable liquid solution. Simply add it to your culture medium, incubate for 30 min and image!
Compatible with 2-photon microscopy
Use it for labeling mitochondria deep in living tissues and multicellular spheroids
Labels mitochondria with reduced membrane potential
Boost your potential to visualize dysfunctional or anaerobic mitochondria for your research on mitochondria-related disorders or anaerobic organisms
Protocol
Dilute
GlowMito 1:2000 in your culture medium
Incubate
for 30 minutes at 37°C
Image
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 & confocal microscopes, flow cytometers Fixable: no
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 one month. For long-term storage, aliquot and store at -20°C.
Outputs:
GlowMito labels the entire mitochondrial network, including those with reduced potential. It is perfectly suitable for:
- Live imaging : study of mitochondrial dynamics (velocity, localization), structural changes, multiplexing (potentiometric dyes, calcium signaling probes, etc).
- Subsequent analysis: flow cytometry, oxygraphy, etc
Although GlowMito accumulation within mitochondria is at least partly driven by membrane potential, some assays are currently being carried out to define the precise correlations between GlowMito signal and mitochondrial membrane potential variations. Therefore, to this day, we cannot guarantee its efficiency for the measure of membrane potential, 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.
Safety datasheet to download here.
Story
"As a chemist, Olivier thrives on finding new synthetic routes giving access to smart molecules. In 2015, he discovered by serendipity a new chemical pathway to reach triaminophenazinium compounds. These molecules were already well known, but their synthetic route drastically limited the nature of the substituents carried by the amine functions. Therefore, Olivier decided to find applications for them in biology and one of the most promising was the mitochondrial tagging. From then on, the collaboration with Guy was obvious. As a specialist of mitochondrial medicine, he would undoubtedly contribute to furthering this marking! Their first approach in fluorescence microscopy made it possible to localize selectively the water-soluble phenaziniums at the level of the mitochondrial network. Together, they characterized the optimal concentration of each triaminophenazinium compound and their kinetics of mitochondrial labeling, further demonstrated their safety on mitochondrial respiration function.
GlowMito was born!"
Olivier Siri and Guy Lenaers
Esther Graudens
New projects at Idylle
The association between Olivier and Guy is synergistic. In addition to their individual talent, it convinced us that their team would drive their tech transfer project further. Our work together was highly fluid and we managed to transfer their technology in a very short time. We are thrilled to launch the GlowMito marker on the research market and I am looking forward to getting the first feedback of use from the early-users!
Combining GlowMito
You may also like to combine GlowMito with other R&D products
Stencell
Use them together and get a live imaging of mitochondria spatial distribution / structure in cell migration
AgarSqueezer
Combine them together to perform live imaging of mitochondria dynamics in response to compression