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MitoFlamma® Green (live)

Product#: RMS1101

$219.36

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MitoFlamma® Green (live)

Cat. No. : RMS1101_20 X 50 UL | RMS1101_1 ML

Description

MitoFlamma® Green is a mitochondria-selective green fluorescent dye that allows to detect mitochondrial morphology in living cells. This unique dye appears to preferentially accumulate in mitochondria. MitoFlamma® green enables researchers to observe mitochondrial activity, localization and abundance as well as monitoring the effect of drugs or other external stimuli on the mitochondrial function. The maxima of Ex/Em values are at 498/583 nm. We offer MitoFlamma® green as an effective mitochondria specific fluorescent probe for various experiments such as immunocytochemistry, in situ hybridization, microplate-based analysis, etc.

Specifications

Fluorophore: MitoFlamma® Green
Application: Cell structure imaging
Labeling to: Mitochondria
Excitation/Emission Max.(nm): 498/583
Molecular weight: 635 g/mol
Appearance: Red Solution
Concentration: 1mM
Solvent: 1% DMSO in PBS or media
Storage conditions: -20 ℃, protect from light

Background

MitoFlamma® Green is a mitochondria-selective green fluorescent dye that allows to detect mitochondrial morphology in living cells. MitoFlamma® Green appears to preferentially accumulate in mitochondria regardless of mitochondrial membrane potential in certain cell types, making it a possible tool for determining mitochondrial mass. MitoFlamma® Green enables researchers to observe mitochondrial activity, localization and abundance as well as monitoring the effect of drugs or other external stimuli on the mitochondrial function. MitoFlamma® Green probes are retained in the mitochondria during the fixation step and after permeabilization with some detergents during subsequent processing steps. After fixation, labeled samples can be applied in a variety of experiments such as immunocytochemistry, in situ hybridization, microplate-based analysis, etc.

Figure 1. Excitation/emission spectra of MitoFlamma® Green

Figure 2. HeLa cell staining with MitoFlamma® Green
Left: DAPI, middle: MitoFlamma Green, right: merged image

Before begining of mitochondrial labeling experiment

Materials and equipment required but not provided

DMSO
PBS buffer or suitable growth medium for live cell imaging
Aldehyde based fixatives such as paraformaldehyde for cell fixation
Aldehyde based detergents such as Trition® X-100
Micropipette
Fluorescence microscope
37 ℃ incubator

Experimental Protocol

Labeling of mitochondria in fixed or suspended cells with MitoFlamma® Green

Prepare the fixed/suspended cells by seeding appropriate cells onto confocal dishes 6 hours before.
Adjust the temperature of the media to 37 ℃.
Prepare a 1 μM labeling MitoFlamma® Green solution by diluting original 1 mM DMSO solution with the media from step 3, *protect from light.
Aspirate the culture medium from the cells grown dishes (for suspended cells, centrifuge the dishes in advance), and add 1 μM labeling MitoFlamma® Green solution to the dishes and incubate for 1 h at 37 ℃ under 5% CO2 in humid condition. *the incubation may be varied for optimal labeling condition.
Aspirate the labeling media, and rinse the cells twice with PBS at 37 ℃.
Analyze stained cells with appropriate equipment

Fixation (continuing from step 5 in labeling experiment)

Fix the cells for 15 min in 4%parnformaldehyde at 37 ℃.
Aspirate the fixative, and rinse the cell twice with PBS.
View the cells using FITC filter set.

Permeabilization (optional)

When permeabilization is needed for subsequent steps such as immunocytochemistry, incubate fixed cells in buffer containing detergent such as 0.2% Triton® X-100.
Following permeabilization, rinse the cells in buffer and proceed with immunocytochemistry procedure.
Alternatively, the cells may be permeabilized by incubating in ice-cold acetone for 5 minutes, and then washed in PBS.
Even when cells are not going to be labeled with an antibody, this acetone-permeabilization step may be useful in improving the signal-to background ratio.