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MemBrite® Fix Cell Surface Staining Kits

Wide choice of dye colors for covalent staining the surface of live cells to conveniently visualize cell boundaries in immunofluorescence experiments.

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MemBrite® Fix Cell Surface Staining Kits
MemBrite® Fix Cell Surface Staining Kits - Image 2MemBrite™ Fix labels the cell surface more evenly than traditional lipophilic membrane stains, and tolerates fixation and permeabilization. HeLa cells were labeled with DiI for 20 minutes at 37°C, or MemBrite™ Fix 568/580 for 5 minutes at 37°C. Cells were then fixed with 4% paraformaldehyde/PBS for 20 minutes at room temperature, followed by permeabilization in 0.1% Triton® X-100 in PBS for 10 minutes at room temperature, or fixed with ice-cold methanol for 5 minutes at -20°C. Because MemBrite™ Fix surface labeling is covalent, it does not redistribute after permeabilization, unlike lipophilic DiI.
MemBrite® Fix Cell Surface Staining Kits - Image 3MemBrite™ Fix labels the cell surface rapidly and specifically, and tolerates fixation and permeabilization. HeLa cells were labeled with MemBrite™ Fix 640/660 for 5 minutes at 37°C or 30 minutes at 4°C. Cells were then fixed with 4% paraformaldehyde, followed by permeabilization with Triton X-100, or fixed with ice cold methanol. Cells labeled at 4°C were also fixed and permeabilized at 4°C.
MemBrite® Fix Cell Surface Staining Kits - Image 4Cells were labeled with the indicated dye for 5 minutes at 37°C, then fixed with 4% paraformaldehyde (PFA) for 20 minutes at room temperature, followed by permeabilization with PBS/0.1% Triton® X-100 for 10 minutes at room temperature.
MemBrite® Fix Cell Surface Staining Kits - Image 5MemBrite™ Fix has excellent tolerance for methanol fixation. Cells were labeled with the indicated dye for 5 minutes at 37°C, then fixed with ice-cold methanol for 5 minutes at -20°C.
MemBrite® Fix Cell Surface Staining Kits - Image 6Labeling with MemBrite™ Fix at low temperature results in highly specific cell surface labeling. Cells were labeled with the indicated dye for 30 minutes at 4°C, then fixed with pre-chilled 4% paraformaldehyde/PBS for 20 minutes at 4°C temperature, followed by permeabilization in 0.1% Triton® X-100 in PBS for 10 minutes at room temperature.
MemBrite® Fix Cell Surface Staining Kits - Image 7S. cerevisiae yeast were labeled separately with MemBrite™ Fix 488/515 (green), MemBrite Fix™ 568/580 (red), and MemBrite Fix™ 640/660 (magenta), then combined and imaged by confocal microscopy.
MemBrite® Fix Cell Surface Staining Kits - Image 8Embryos of the tunicate Phallusia mammillata were stained using MemBrite® Fix 543/560 (red) at stage 12: mid gastrula, followed by downstream HCR-FISH. The embryos were mounted with DAPI (blue) and imaged using confocal microscopy. Left: plane through the animal hemisphere of the embryo. Center: the center of the embryo. Right: plane through the vegetal hemisphere of the embryo with the FISH-labeled RNA transcript (green). Scale bar is 100 um. Credit: Ingrid Mormin, research assistant, Patrick Lemaire’s lab, CRBM-CNRS UMR5237.
MemBrite® Fix Cell Surface Staining Kits - Image 9Embryo of the tunicate Phallusia mammillata stained with MemBrite® Fix 543/560 (red) at stage 12: mid gastrula and mounted with DAPI (blue). Images progress through the z-stack from the animal hemisphere to the vegetal hemisphere of the embryo. Credit: Ingrid Mormin, research assistant, Patrick Lemaire’s lab, CRBM-CNRS UMR5237.
MemBrite® Fix Cell Surface Staining Kits - Image 10PEG-enriched HeLa-derived EVs were stained with 1X MemBrite® Fix 405/430 (right). Specific staining was seen, compared with MemBrite® Fix 405/430 in buffer (left). EVs were detected on a Cytoflex LX flow cytometer in the Pacific Blue channel.
MemBrite® Fix Cell Surface Staining Kits - Image 11SEC-purified MCF-7-derived exosomess were stained with 1X MemBrite® Fix 405/430 (right). Specific staining was seen, compared with MemBrite® Fix 405/430 in buffer (left). EVs were detected on a Cytoflex LX flow cytometer in the Pacific Blue channel.
MemBrite® Fix Cell Surface Staining Kits - Image 12Jurkat cells were stained with 1X MemBrite® Fix 488/515 dye , after pre-treatment with the Pre-Staining Solution. Cells were washed and then labeled with an antibody conjugate against the indicated target protein. The antibody staining was analyzed by flow cytometry in the APC channel. Staining the cells with MemBrite® Fix 488/515 prior to antibody labeling had no effect on the antibody signal (middle panels, filled red peaks), compared to cells stained with antibody alone (top panels, unfilled peaks). Unstained cells are shown in gray in the bottom panels.
MemBrite® Fix Cell Surface Staining Kits - Image 13Jurkat cells were stained with 1X MemBrite® Fix dye either with the recommended Pre-Staining treatment (darker colored peaks) or without Pre-Staining treatment (lighter colored peaks). Gray peaks represent unstained cells. Treating cells with the Pre-Staining Solution increases the brightness of MemBrite® Fix staining.

Product Description

MemBrite® Fix Cell Surface Staining Kits provide a convenient way to visualize cell boundaries in multicolor staining experiments. Staining is rapid and uniform, with a wide choice of colors. MemBrite® Fix dyes covalently label the surface of live cells. The staining withstands fixation and permeabilization for subsequent immunofluorescence staining.

  • Rapid, uniform live cell surface staining
  • Covalent labeling can be fixed & permeabilized for IF staining
  • Stain mammalian cells, yeast, or gram-positive bacteria
  • Choose from 12 bright & photostable dye colors from blue to near-IR
  • Dye options for super-resolution and other specialized imaging applications

Click to enlarge

Fixable Cell Surface Stains

MemBrite® Fix Stains are novel reactive fluorescent dye stains that react irreversibly with cell surface proteins, for staining that can withstand formaldehyde or alcohol fixation, and detergent permeabilization. Unlike lectins such as WGA, which bind specific targets that may vary between cell types, MemBrite® Fix dyes react widely with cell surface proteins. MemBrite® Fix staining is rapid and non-toxic to cells, and because MemBrite® Fix dyes are highly water soluble, they stain cells much more evenly than traditional lipophilic membrane dyes like DiO, DiI, PKH, Vybrant®, or CellMask™.

MemBrite® Fix staining kits also can be used to stain yeast and gram-positive bacteria, but not gram-negative bacteria. See our Cellular Stains Table for more information on how our dyes stain various organisms. MemBrite® Fix 405/430 has been validated for staining of extracellular vesicles (EVs) and exosomes.

MemBrite® Fix Staining Kits belong to Biotium’s line of novel cell surface stains that include CellBrite® Fix Membrane Stains. CellBrite® Fix Membrane Stains are fluorogenic dyes that rapidly accumulate in the plasma membrane, where they react covalently with the cell surface. CellBrite® Fix stains require only a single staining step compared to MemBrite® Fix staining, which is a two-step protocol. On the other hand, MemBrite® Fix dyes are available with a wider selection of colors. MemBrite® dyes do not associate with lipids in membranes, and consequently have lower cytoplasmic background after detergent permeabilization compared to CellBrite® Fix.

Selecting a MemBrite® Fix Dye

Several MemBrite® Fix dyes have been validated in super-resolution imaging applications or 2-photon microscopy. MemBrite® Fix-ST dyes are recommended for super-resolution imaging by STORM. MemBrite® Fix or MemBrite® Fix-ST dyes can be used for standard microscopy applications; however, MemBrite® Fix dyes are generally more photostable than MemBrite® Fix-ST dyes. See the MemBrite® Fix Product Table below for details.

Tips for Success

Note that MemBrite® Fix dye stain dead cell more intensely than live cells. With prolonged dye incubation, or if cells are cultured after staining, the dye also will be internalized by endocytosis, resulting in labeling of intracellular vesicles. Please see our Tech Tip: Five Steps for Success with Membrane and Surface Stains for tips on staining and imaging (step 5) with MemBrite® Fix.

Find the Right Stain for Your Application

MemBrite® Fix dyes must be used to stain live cells before fixation. They cannot be used to stain cells that are already fixed (the dyes primarily label intracellular membranes in fixed cells). Our original CellBrite® Cytoplasmic Membrane Dyes can be used to stain cells after fixation and permeabilization, see our Tech Tip: Combining Lipophilic Membrane Dyes with Immunofluorescence. To find the right stain for your application, see our Membrane & Cell Surface Stains Comparison, or download our Membrane & Surface Stains Brochure.

MemBrite® Fix dyes are designed to be fixed shortly after staining. With prolonged dye incubation, or if cells are cultured after staining, the dyes will be internalized by endocytosis, resulting in labeling of intracellular vesicles. By 24 hours after staining, most of the dye will be localized inside the cell, not on the cell surface. For long-term visualization of cell boundaries in culture, we recommend our CellBrite® Steady Membrane Staining Kits. These kits include unique fluorescent membrane probes that retain cell surface staining in live cell cultures for 24 hours or longer. The kits also include an optional CellBrite® Steady Enhancer solution which masks intracellular signal for even greater specificity of cell boundaries.

Watch our video where Technical Applications Scientist II, Jacqueline Steenhuis PhD answers your top questions about Biotium’s various membrane stains for fluorescence microscopy.

For additional support or product recommendations, contact us at techsupport@biotium.com.

MemBrite® Fix Cell Surface Staining Kits

Catalog numberSize1Dye2Spectrally
similar to		Specialized
applications
30092-T100 reactionsMemBrite® Fix 405/430Alexa Fluor® 405
CellBrite® Blue		SIM
exosome staining
30092500 reactions
30093-T100 reactionsMemBrite® Fix 488/515FITC
Alexa Fluor® 488
DiO
CellBrite® Green		STED
TIRF
2-photon microscopy
30093500 reactions
30094-T100 reactionsMemBrite® Fix 543/560TAMRA
Cy®3
Alexa Fluor® 546
DiI
CellBrite® Orange		N/A
30094500 reactions
30095-T100 reactionsMemBrite® Fix 568/580Alexa Fluor® 568
Rhodamine Red
DiI
CellBrite® Orange		SIM
STORM
TIRF
30095500 reactions
30096-T100 reactionsMemBrite® Fix 594/615Texas Red®
Alexa Fluor® 594		2-photon
microscopy
30096500 reactions
30097-T100 reactionsMemBrite® Fix 640/660Cy®5,
Alexa Fluor® 647
DiD
CellBrite® Red		FLImP
SIM
TIRF
30097500 reactions
30098-T100 reactionsMemBrite® Fix 660/680Alexa Fluor® 660N/A
30098500 reactions
30099-T100 reactionsMemBrite® Fix 680/700Cy®5.5
Alexa Fluor® 680
IRDye® 680LT
CellBrite® NIR 680		STORM3
Single-molecule imaging
STED
2-photon microscopy
30099500 reactions
30101-T100 reactionsMemBrite® Fix-ST 650/665Cy®5
Alexa Fluor® 647
DiD
CellBrite® Red		STORM
30101500 reactions
30102-T100 reactionsMemBrite® Fix-ST 667/685Alexa Fluor® 660STORM
30102500 reactions
30103-T100 reactionsMemBrite® Fix-ST 681/698Cy®5.5
Alexa Fluor® 680
IRDye® 680LT
CellBrite® NIR 680		Single-molecule imaging
STORM
30103500 reactions
30104-T100 reactionsMemBrite® Fix-ST 755/777Alexa Fluor® 750
Cy®7
DyLight® 750		STORM
30104500 reactions
FLImP: Fluorophore localization imaging with photobleaching; SIM: Structured illumination microscopy; STED: Stimulated emission depletion; STORM: Stochastical optical reconstruction microscopy; TIRF: Total internal reflection fluorescence

1. Kit size based on 200 uL reaction volume, actual number of reactions will depends on staining volume used.
2. MemBrite™ Fix dyes are named for their Abs/Em maxima. Download the Product Information Sheet for spectra.
3. MemBrite® Fix-ST 681/698 dye is reported to have better performance in STORM imaging than MemBrite® Fix 680/700 dye.

Alexa Fluor & Texas Red are registered trademarks of Thermo Fisher Scientific; Cy Dye is a registered trademark of Cytiva; IRDye is a registered trademark of LI-COR Bioscience.

Vybrant is a registered trademark and CellMask is a trademark of Thermo Fisher Scientific.

References

Download a list of curated CellBrite® and MemBrite® references.

Product Attributes

Dye
MemBrite® Fix 405/430, MemBrite® Fix 488/515, MemBrite® Fix 543/560, MemBrite® Fix 568/580, MemBrite® Fix 594/615, MemBrite® Fix 640/660, MemBrite® Fix 660/680, MemBrite® Fix 680/700, MemBrite® Fix-ST 650/665, MemBrite® Fix-ST 667/685, MemBrite® Fix-ST 681/698, MemBrite® Fix-ST 755/777
Size
100 labelings, 500 labelings
Probe cellular localization
Membrane/cell surface, Membrane/vesicular
Cell permeability
Membrane impermeant
For live or fixed cells
Covalent & fixable stains, For live/intact cells
Assay type/options
Co-cultures, Long term staining (24-72h)
Colors
Blue, Green, Red, Far-red, Near-infrared
Fixation options
Fix after staining (formaldehyde), Fix after staining (methanol), Permeabilize after staining

Documents, Protocols, SDS and COA

Citations

Download a list of curated CellBrite® and MemBrite® references.

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FAQs

CellBrite® & MemBrite® Membrane & Cell Surface Stains

While CellBrite® Cytoplasmic Membrane Dyes can stain formaldehyde-fixed cells, they generally do not give good results in cryosections, possibly because the cell membrane integrity is disrupted, exposing other membrane structures to the dyes. Some customers have reported success using these dyes with vibratome sections.

CellBrite® Cytoplasmic Membrane Dyes are not suitable for membrane staining in FFPE samples as membrane lipids are extracted during the dewaxing and rehydration process. Similarly, acetone or methanol fixation of cryosections will extract lipids, leading to poor staining.

CellBrite® Fix, MemBrite® Fix, and CellBrite® Steady are recommended for use on live cells only. In fixed cells or sections they will label intracellular structures.

In some tissue types, lectins such as CF® Dye WGA Conjugates, CF® Dye Concanavalin A Conjugates, or CF® Dye PNA Conjugates may be useful for staining cell boundaries in FFPE or frozen sections. However, the staining pattern of lectins is highly dependent on cell and tissue type, so we recommend consulting the literature before trying these stains for your tissue of interest.

Alternatively, immunostaining using cell surface-specific antibodies could be done.

So far we have not found a universal plasma membrane stain for tissue sections. This is an application of interest to us and our customers, so we are working to find new solutions.

 

CellBrite® Cytoplasmic Membrane Dyes are too prone to aggregation to efficiently stain EVs. Some of the CellBrite® Fix, MemBrite® Fix, and CellBrite® Steady dye options have been reported for this application, however we do not recommend them. For optimal staining of exosome membranes we recommend our ExoBrite™ True EV Membrane Stains, which are novel lipophilic membrane dyes specifically designed and optimized for efficient staining of EV membranes with minimal dye aggregation. See our Extracellular Vesicle Research page for more information about our complete line of EV stains and antibodies.

Lipophilic carbocyanine dyes like CellBrite® and CellBrite® NIR Cytoplasmic Membrane Dyes can be used to stain formaldehyde-fixed cells. Fixation with methanol or other solvents is not recommended. Permeabilization with detergents after staining will extract the dyes and alter the staining pattern. However, we’ve seen good results when formaldehyde-fixed cells are permeabilized before staining with CellBrite® dyes (See Tech Tip: Combining Lipophilic Membrane Dyes with Immunofluorescence).

CellBrite® Fix, MemBrite® Fix, and CellBrite® Steady cannot be used to stain the plasma membrane of fixed samples; these dyes will primarily stain intracellular structures in cells that are already fixed.

To find the right dye for your workflow, see our Comparison of Membrane & Cell Surface Stains, or download our Membrane & Surface Stains Brochure.

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