Shop All Cell Stains

FM™ 5-95 (N-(3-Trimethylammoniumpropyl)-4-(6-(4-(Diethylamino)phenyl)hexatrienyl)Pyridinium Dibromide) Invitrogen™

The increasing number of successful applications for our FM dyes has prompted us to synthesize FM 5-95, a slightly less lipophilic analog of FM 4-64 (T-3166, T-13320) with essentially identical spectroscopic properties.

Filmtracer™ LIVE/DEAD™ Biofilm Viability Kit Invitrogen™

The FilmTracer™ LIVE⁄DEAD Biofilm Viability kit provides a two-color fluorescence assay of bacterial viability, based on membrane integrity, that has proven usefule for a diverse array of bacterial genera including those growing in biofilm communities. The LIVE⁄DEAD Biofilm Viability kit utilizes mixtures of our SYTO® 9 green fluorescent nucleic acid stain and the red-fluorescent nucleic acid stain, propidium iodide. These stains differ both in their spectral characteristics and in their ability to penetrate healthy bacterial cells. When used alone, the SYTO® 9 stain generally labels all bacteria in a population—those with intact membranes and those with damaged membranes. In contrast, propidium iodide penetrates only bacteria with damaged membranes, causing a rduction in the SYTO® 9 stain fluorescence with both dyes are present. Thus, with an appropriate mixture of the SYTO™ 9 and propdium iodide stains, bacteria with intact cell membranes stain fluorescent green, whereas bacteria with damaged membranes stain fluorescent red.

FM™ 4-64 Dye (N-(3-Triethylammoniumpropyl)-4-(6-(4-(Diethylamino) Phenyl) Hexatrienyl) Pyridinium Dibromide) Invitrogen™

The styryl dye FM 4-64 has been reported to selectively stain yeast vacuolar membranes with red fluorescence (excitation/emission maxima ~515/640 nm). This lipophilic dye is an important tool for visualizing vacuolar organelle morphology and dynamics, for studying the endocytic pathway and for screening and characterizing yeast endocytosis mutants. FM 4-64 is also available in special packaging with 10 vials of 100 µg each (T-13320).

NucBlue™ Live ReadyProbes™ Reagent (Hoechst 33342) Invitrogen™

Hoechst 33342 is a popular cell-permeant nuclear counterstain that emits blue fluorescence when bound to DNA. With NucBlue® Live ReadyProbes® Reagent we have formulated this classic stain in a room temperature-stable solution that is provided in a convenient-to-use dropper bottle. Just tip and drip two drops per ml to stain your cells.

Also available: Hoechst 33342, trihydrochloride, trihydrate at 12.5X concentration (40.6 µM) in water.

• No need to dilute, weigh, or pipette
• Convenient dropper bottle—just use two drops per mL
• Stable at room temperature—keep handy at your scope or cell culture area
• Excited by UV light and emits blue fluorescence at 460 nm when bound to DNA nm

See other ReadyProbes® reagents for cell staining
See other nuclear stains for imaging

Cell imaging applications
The spectral properties of Hoechst 33342 (2'-[4-ethoxyphenyl]-5-[4-methyl-1-piperazinyl]-2,5'-bi-1H-benzimidazole), including a large Stokes shift, make it ideal for use with green (Alexa Fluor® 488, FITC, GFP) and red (Alexa Fluor® 594, Texas Red®, rhodamine, mCherry, mKate-2) fluorophores in multicolor experiments. Because of its high affinity to DNA, Hoechst 33342 is also frequently used in cell counting, cell cycle, and cell replication studies to distinguish condensed nuclei in apoptotic cells, for cell-cycle studies in combination with Click-iT® EdU or BrdU staining, as a nuclear segmentation tool in high content imaging analysis, and to sort cells based on their DNA content.

Suggestions for use
NucBlue® Live Cell Stain may be added directly to cells in full media, or buffer solutions.
• In most cases 2 drops/ml and an incubation of 15 to 30 minutes will give bright nuclear staining; however, optimization may be needed for some cell types, conditions, and applications. In such cases simply add more, or fewer, drops until the optimal staining intensity is obtained. In most cases, staining intensity increases with time if cells are not washed prior to imaging.
• NucBlue® Live Cell Stain is excited by UV light at 360 nm when bound to DNA, with an emission maximum at 460 nm. It is detected through a blue/cyan filter, such as a DAPI filter, blue GFP filters, or the Semrock BrightLine® Alexa Fluor® 350 Dye filter set.

SYTO™ 60 Red Fluorescent Nucleic Acid Stain - 5 mM Solution in DMSO Invitrogen™

The cell-permeant SYTO 60 red fluorescent nucleic acid stain exhibits bright, red fluorescence upon binding to nucleic acids. Because the staining pattern of the SYTO dyes in live cells may vary between cell types, we offer the SYTO Red Fluorescent Nucleic Acid Stain Sampler Kit (S-11340) to enable researchers to find the most appropriate red-fluorescent SYTO stain for their system.

JC-1 Dye (Mitochondrial Membrane Potential Probe) Invitrogen™

JC-1 is a novel cationic carbocyanine dye that accumulates in mitochondria. The dye exists as a monomer at low concentrations and yields green fluorescence, similar to fluorescein. At higher concentrations, the dye forms J-aggregates that exhibit a broad excitation spectrum and an emission maximum at ~590 nm. These characteristics make JC-1 a sensitive marker for mitochondrial membrane potential. Another dye with similar characteristics is JC-9 (D-22421).

MitoTracker™ Red CM-H2Xros - Special Packaging Invitrogen™

MitoTracker Red CM-H2XRos is a reduced, nonfluorescent version of MitoTracker Red (M-7512) that fluoresces upon oxidation. This dye also stains mitochondria in live cells and its accumulation is dependent upon membrane potential. The dye is well-retained after aldehyde fixation.

NucBlue™ Fixed Cell ReadyProbes™ Reagent (DAPI) Invitrogen™

DAPI is a commonly used nuclear counterstain for fixed cells that emits blue fluorescence when bound to DNA. With NucBlue® Fixed Cell ReadyProbes® Reagent, we have formulated a high-purity form of this classic stain in a room temperature-stable solution that is provided in a convenient-to-use dropper bottle. Just tip and drip two drops per ml to stain your cells.

Also available: DAPI (4',6-diamidino-2-phenylindole, dihydrochloride) at 12.5X concentration (5 µM) in water.

• No need to dilute, weigh, or pipette
• Convenient dropper bottle—just use two drops per mL
• Stable at room temperature—keep handy at your scope or cell culture area• DAPI is excited with UV light and detected through a blue/cyan filter

See other ReadyProbes® reagents for cell staining
See other nuclear stains for imaging

Cell imaging applications
DAPI is a classic fluorescent dye used extensively for nuclear staining of fixed cells. In fluorescence microscopy, DAPI is excited with UV light and detected through a blue/cyan filter (Figure 1)..

Suggestions for use
In most cases 2 drops/ml of NucBlue® Fixed Cell Stain and an incubation of 15 to 30 minutes will produce bright nuclear staining; however, optimization may be needed for some cell types, conditions, and applications. In such cases, simply add more, or fewer, drops until the optimal staining intensity is obtained.
• NucBlue® Fixed Cell Stain is excited by UV light at 360 nm when bound to DNA, with an emission maximum at 460 nm. It is detected through a blue/cyan filter, such as a DAPI filter, blue GFP filters, or the Semrock BrightLine® Alexa Fluor® 350 Dye filter set.
• As a preferred blue nuclear stain in fixed cell imaging experiments, NucBlue® Fixed Cell Stain is ideal for use with antibody-based applications.

TOTO™-1 Iodide (514/533) - 1 mM Solution in DMSO Invitrogen™

TOTO®-1 iodide is a cell-impermeant, high-affinity nucleic acid stain based upon a symmetric cyanine dye dimer that is essentially nonfluorescent in the absence of nucleic acids that exhibits excitation/emission maxima ~514/533 nm when bound to nucleic acids.

LysoSensor™ Yellow/Blue dextran, 10,000 MW, Anionic, Fixable Invitrogen™

LysoSensor Yellow/Blue dextran is a ratiometric probe that can be used to measure the pH of acidic organelles, such as lysosomes. This dextran conjugate is taken up by the cells through endocytosis. The LysoSensor dye produces blue fluorescence in neutral environments, but changes to yellow fluorescence in more acidic environments. The pKa of LysoSensor Yellow/Blue dextran is ~3.9.

DRAQ5™ Fluorescent Probe Solution (5 mM) Thermo Scientific™

The Thermo Scientific DRAQ5™ Fluorescent Probe is a far-red DNA stain for live or fixed cells and easy multiplexing with other fluorophores, such as cells expressing green fluorescent protein (GFP) fusion proteins.

DRAQ5™ Fluorescent Probe is a far-red DNA stain for fluorescent cellular imaging applications with live or fixed cells. Because of its far-red excitation and emission, the DRAQ5 Stain can be multiplexed with many other fluorophores. It is commonly chosen to counterstain nuclei and measure DNA content in cell proliferation studies involving cells expressing green fluorescent protein (GFP) fusion proteins. DRAQ5 Fluorescent Probe is compatible with many existing protocols across a wide range of instrumentation platforms.

Features of the DRAQ5 Fluorescent Probe:

DRAQ5 dye—a red-to-infrared fluorescent stain specific for DNA (nuclei and DNA quantitation)
Convenient—provided as easy-to-use DRAQ5 dye solutions (5 mM)
Cellular imaging—lipophilic and membrane permeable dye is effective for live-cell and fixed-cell staining and quantitation of DNA content
Counterstain—ideal for use alongside other fluorophore reporters for fluorescence microscopy or high-content screening (HCS)

Applications:
• Flow cytometry
• Flow and laser scanning cytometry
• Live cell imaging
• Cell based assays

Properties of DRAQ5 Fluorescent Dye
• Structure: Proprietary
• Alternative names: Anthraquinone compound
• Chemical name(s): 1,5-bis{[2-(di-methylamino) ethyl]amino}-4, 8-dihydroxyanthracene-9,10-dione
• Molecular formula: Not available
• Molecular weight: 412.54
• Excitation wavelength: 647 nm line optimal (Exλmax = 646 nm)
• Emission wavelength: > 665 nm to infra-red > 800 nm (Emλmax = 681 nm/697 nm intercalated with dsDNA)
• Extinction coefficient: Not available
• CAS #: Not available
• Purity: > 99%
• Solubility: Soluble at working concentrations in water and typical buffers
• Reactive groups: None; binds DNA A-T minor groove, mimicking histone 2b distribution, resulting in 16 nm red-shift

Fluorescence cell-based assays, such as flow cytometry, in-cell ELISA, fluorescence microscopy and high-content imaging require a fluorescent label to identify individual cells. When using multiple fluorescent probes to detect different cellular targets or activities, each probe must have a fluorescent spectrum different than the other probes. The blue-fluorescent DNA-binding probes, Hoechst and DAPI are frequently used; however, these probes cannot be used when UV illumination is unavailable or other blue-emitting fluorescent probes are used. Therefore, nuclear probes that emit in a color other than blue are useful for cell identification and counting, and for determining nuclear morphology and DNA content.

DRAQ5 Fluorescent Probe emits in the far-red region, is lipophilic and crosses cell and nuclear membranes in live and fixed cells and tissues for rapid DNA staining. This stain is water-soluble, supplied ready to use and does not require RNase, cell lysis, or a washing step, making it compatible with automation. Because DNA staining is stoichiometric, the DRAQ5 Fluorescent Probe can be used for DNA content analysis in cell proliferation studies. DRAQ5 Fluorescent Probe is usually added as the last stain in a labeling procedure because no washing is required. Alternatively, add this probe in assay medium for a live cell assay.

SYTO™ 61 Red Fluorescent Nucleic Acid Stain - 5 mM Solution in DMSO Invitrogen™

The cell-permeant SYTO 61 red fluorescent nucleic acid stain exhibits bright, red fluorescence upon binding to nucleic acids. Because the staining pattern of the SYTO dyes in live cells may vary between cell types, we offer the SYTO Red Fluorescent Nucleic Acid Stain Sampler Kit (S-11340) to enable researchers to find the most appropriate red-fluorescent SYTO stain for their system.

MitoTracker™ Green FM - Special Packaging Invitrogen™

MitoTracker Green FM is green-fluorescent mitochondrial stain which appears to localize to mitochondria regardless of mitochondrial membrane potential. The dye will stain live cells but is not well-retained after aldehyde fixation.

TO-PRO™-1 Iodide (515/531) - 1 mM Solution in DMSO Invitrogen™

One of our carbocyanine monomers, TO-PRO®-1 iodide is a useful counterstain and dead cell indicator, with excitation/emission maxima ~515/531 nm.

CellMask™ Green Plasma Membrane Stain Invitrogen™

The CellMask™ Green Plasma Membrane Stain enables fast and uniform labeling of the plasma membrane in almost all cell types. This stain works rapidly and can be detected using a standard FITC filter set with any imaging instrument. Depending upon the cell type and experimental conditions, the plasma membrane staining will last for 60–90 minutes without detectable internalization, providing enough time for most live cell dynamic studies. The stain survives fixation, but not permeabilization.

Features of the CellMask™ Green Plasma Membrane Stain include:

• Sensitivity—specific and rapid plasma membrane staining
• Excellent retention—staining lasts for 60–90 minutes
• Emission/excitation (522/535 nm) works with standard FITC settings

Drawbacks of Other Methods
The plasma membrane is a convenient marker of cell boundaries and as such, a number of probes have been used for staining of the membrane. Typically, dyes of a lipophilic nature are used; however, they internalize rapidly, offering a very narrow window for imaging. Fluorescently labeled lectins, such as wheat germ agglutinin, have also been employed as plasma membrane stains. Conjugated lectins depend on cell surface sugars for staining and as a result, stain inconsistently with variation across cell types. Robust plasma membrane staining is important for a range of applications including translocation assays, plasma membrane dynamics, and as a general tool for cell identification in traditional and automated imaging and analysis.

How CellMask™ Plasma Membrane Stains Work
CellMask™ Green Plasma Membrane Stain is designed to deliver uniform staining of the plasma membrane across a wide variety of mammalian cell types and is slow to internalize, particularly compared to traditional approaches such as DiI, DiO, and labeled wheat germ agglutinin. CellMask™ plasma membrane stains are amphipathic molecules containing a lipophilic moiety for excellent membrane loading and a negatively charged hydrophilic dye for 'anchoring' of the probe in the plasma membrane. While the CellMask™ plasma membrane stains provide ample opportunity for live cell imaging, the staining pattern is also maintained after fixation with formaldehyde, enabling additional multiparametric imaging options. However, staining with CellMask™ plasma membrane stains does not survive detergent extraction and therefore cannot be used in conjunction with probes that require permeabilization.
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