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Membrane &
Cell Surface Stains

Find the right dye for your application

Find a Membrane or Surface Stain for Your Workflow

Researchers frequently wish to label the cell surface or plasma membrane to visualize cellular boundaries or cell morphology in multi-color fluorescence imaging. Biotium offers a number of traditional and novel stains that are designed for convenient labeling of live cells, fixed cells, or for fixation and subsequent immunostaining. See below to find the right stain for your application.

A. Stain live cells

1. Stain live cells

↓

2. Wash1

↓

3. Image live cells2

Recommended products:
CellBrite® Cytoplasmic Membrane Dyes
CellBrite® Steady Membrane Stains

1 Washing is optional for CellBrite® Steady Stains when imaged by confocal.

2 CellBrite® Cytoplasmic Membrane Dyes stain cells stably for tracking experiments, but are rapidly internalized. CellBrite® Steady Kits can be used to image cell surface in culture for days.

B. Stain then fix

1. Stain live cells

↓

2. Fix cells

↓

3. (Optional) Proceed to
additional staining

Recommended products:
CellBrite® Cytoplasmic Membrane Dyes3
CellBrite® Fix Membrane Stains
MemBrite® Fix Membrane Stains
WGA Conjugates
ConA Conjugates

 

3 Formaldehyde fixation recommended for CellBrite® Cytoplasmic Membrane Dyes.

C. Stain, fix, & permeabilize

1. Stain live cells

↓

2. Fix cells

↓

3. Permeabilize cells

↓

4. (Optional) Proceed to
additional staining

Recommended products:
CellBrite® Fix Membrane Stains
MemBrite® Fix Membrane Stains
WGA Conjugates
ConA Conjugates

 

D. Fix, then stain

1. Fix cells

↓

2. Stain cells

Recommended products:

CytoLiner™ Fixed Cell Membrane Stains3
CellBrite® Cytoplasmic Membrane Dyes3
WGA Conjugates
ConA Conjugates

3 Formaldehyde fixation recommended for CytoLiner™ Fixed Cell Membrane Stain and CellBrite® Cytoplasmic Membrane Dyes

Compare Membrane and Cell Surface Stains

Stain Dye transfer
between cells		 Non-toxic, stable1 Stains
yeast		 Stains
bacteria		 Stain after fixing2 Fix after
staining		 Tolerates detergent /
MeOH fix?		 Color
selection		 Pros & Cons
CytoLiner™ Fixed Cell Membrane Stains Yes Yes N/A N/A Yes3 Yes5 No6 5 colors
(blue to NIR)		 Pro: Specifically formulated for bright & uniform membrane staining in fixed cells
Pro: Suitable for antibody co-staining
Con: High background in live cells
CellBrite® Steady
Membrane Staining Kits		 Yes Yes No No No No No 5 colors
(blue to NIR)		 Pro: Image live cell surface for days
Pro: Enhancer masks intracellular staining
Pro: Fast, even staining in medium
Con: Does not tolerate fixation
CellBrite® Cytoplasmic
Membrane Dyes		 Minimal Yes No No Yes3 Yes3 No 4 colors
(blue to far-red)		 Pro: Fix before or after staining3
Con: Uneven live cell staining
Con: Poor tolerance for MeOH/detergent
CellBrite® NIR
Near-Infrared Cytoplasmic Membrane Dyes		 Minimal Yes No No Yes Yes No 4 NIR
dye colors
CellBrite® Fix
Membrane Stains		 Minimal Yes Yes Yes No Yes Yes Green,
red, &
far-red		 Pro: Rapid, uniform staining
Pro: Wash after stain optional
Pro: Fix & perm cells for IF
Pro: Stains yeast & bacteria
Con: Can’t stain fixed cells
MemBrite® Fix
Cell Surface Staining Kits		 Minimal Yes Yes Gram+
only		 No Yes Yes 8 colors
(blue to NIR)		 Pro: Rapid, uniform staining
Pro: Fix & perm cells for IF
Pro: Great color selection
Pro: Stains yeast
Con: Can’t stain fixed cells
CF® Dye
WGA Conjugates		 Possible Possibly toxic4 Bud scars Fluorescent
Gram stain		 Yes Yes Yes 13 colors
(UV to NIR)		 Pro: Fix before or after staining
Pro: Fluorescent Gram stain
Pro: Yeast bud scar stain
Pro: Great color selection
Con: May vary among cell types
CF® Dye
Con A Conjugates		 Possible Possibly toxic4 Yes Reported to
stain biofilms		 Yes Yes Yes 10 colors
(UV to NIR)		 Pro: Fix before or after staining
Pro: Great color selection
Pro: Stains yeast
Con: May vary among cell types
1. Membrane and cell surface labels will be internalized by endocytosis in live cells over time.
2. Intracellular membranes may be labeled in fixed cells.
3. Formaldehyde fixation only.
4. Lectins may be toxic or stimulatory to live cells depending on cell type.
5. CytoLiner™ Fixed Cell Membrane Stains may have higher background if used to stain live cells before fixation than when used to stain cells after fixation.
6. Mild permeabilization with 0.1% Triton® X-100 is recommended, see product information sheet for more info.

Live MCF7 cells were stained with CellBrite® Steady 650 plus Enhancer in complete culture medium for 48 hours and imaged by confocal microscopy without a wash step.
HeLa cells were fixed with formaldehyde, then stained with the CellBrite® Blue Cytoplasmic Membrane Staining Kit, washed, and imaged by confocal microscopy.
HeLa cells stained with CellBrite® Fix 555 (red), then MeOH-fixed and stained with CF®640R Mitochondrial Marker mAb clone 113-1 (cyan) and NucSpot® 470 (green).
S. cerevisiae were stained separately with MemBrite® Fix 488/515 (green), MemBrite® Fix 568/580 (red), and MemBrite® Fix 640/660 (magenta) then mixed together before imaging.
Live HeLa cells were stained with CF®594 Concanavalin A (Con A, red) and Hoechst 33342 (blue) for 10 minutes at 37°C in HBSS, then washed and imaged by confocal microscopy.
MemBrite® Fix labels cells more evenly than traditional membrane stains, and tolerates permeabilization. HeLa cells were labeled with DiI or MemBrite® Fix 568/580, then fixed/permeabilized with formaldehyde followed by Triton® X-100, or fixed with methanol. Unlike DiI, MemBrite® Fix labeling is covalent, so it does not redistribute or wash off after permeabilization.
MemBrite® Fix kits offer a wide selection of dye colors, with excellent tolerance for common fixatives like PFA and methanol. 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.

Stain Cell Surfaces in Whole Organisms

MemBrite® Fix stains have been used to label cell surfaces in whole organisms. In the following figures provided by Patrick Lemaire’s lab of CRBM (Centre de Recherche en Biologie cellulaire de Montepellier), MemBrite® Fix 543/560 was used to stain whole embryos of the tunicate Phallusia mammillata followed by downstream hybridization chain reaction fluorescent in situ hybridization (HCR-FISH) for detection of a RNA transcript. Cell boundaries were clearly labeled through the entirety of the Z-stack from the animal hemisphere to the vegetal hemisphere of the embryo.

Embryos 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.
Embryo 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.

Download our Membrane and Cell Surface Stains Brochure. Also see our complete selection of Cellular Stains, or download our Cellular Stains Brochure.

Faq

CellBrite® & MemBrite® Membrane & Cell Surface Stains
What is the difference between CellBrite®, CellBrite® NIR, CellBrite® Fix, and MemBrite® Fix?
CellBrite® Cytoplasmic Membrane Stains are lipophilic dyes for simple, non-toxic, stable labeling of membranes in live or fixed cells. Cells can be fixed with formaldehyde before or after CellBrite® staining. But the staining has poor tolerance for permeabilization or methanol fixation, so CellBrite® staining is not easily combined with intracellular immunofluorescence (IF) staining. The dyes also do not stain bacteria or yeast. CellBrite® NIR dyes are CellBrite® dyes with near-infrared fluorescence compatible with small animal NIR imaging systems. CellBrite® Fix and MemBrite® Fix stains were developed to overcome some of these shortcomings. They are novel covalent stains that can be fixed and permeabilized for IF staining. CellBrite® Fix Membrane Stains are fluorogenic reactive membrane dyes that rapidly accumulate at the plasma membrane. When they incorporate into lipids, they become fluorescent, and at the same time react covalently with membrane proteins for stable labeling. Staining takes only 15 minutes in a single step with no wash. CellBrite® Fix stains mammalian cells, yeast, and bacteria. MemBrite® Fix Cell Surface Stains do not bind lipids, but label cell surface proteins. MemBrite® Fix requires a two-step staining protocol with washing, but offers a more extensive choice of dye colors than CellBrite® Fix. MemBrite® Fix also can be used to stain yeast. But unlike original CellBrite® dyes and lectins, CellBrite® Fix and MemBrite® Fix cannot be used on cells that are already fixed. To select a dye that’s right for your application, see our Membrane and Cell Surface Stains Comparison, or download our Membrane & Surface Stains Brochure.


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Can CellBrite® or MemBrite® Stains be used to stain exosomes/extracellular vesicles?
CellBrite® Cytoplasmic Membrane Dyes do not efficiently stain EVs. Some of the CellBrite® Fix, MemBrite® Fix, and CellBrite® Steady dye options can be used for this application. However, 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


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What is the difference between CellBrite® dyes and PKH dyes?
Biotium’s CellBrite® Cytoplasmic Membrane Dyes are dye delivery solutions that can be added directly to normal culture media to uniformly label suspended or adherent cells in culture. The PKH dyes are structurally related dyes for cell membrane labeling. But unlike CellBrite®, labeling with PKH dyes requires multiple steps and subjects cells to an iso-osmotic mannitol loading medium that can negatively affect cell membrane integrity and viability. CellBrite® Fix dyes also feature rapid and simple labeling in isotonic buffer. Cells can be fixed and permeabilized after labeling with CellBrite® Fix, unlike original CellBrite® dyes or PKH dyes, which don’t tolerate detergent.


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