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CellBrite™ Steady Membrane Staining Kits

CellBrite™ Steady Membrane Staining Kits

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CellBrite™ Steady Membrane Staining Kits
Live MCF7 cells stained with 1X CellBrite™ Steady 650 or CellBrite™ Steady 685 dye, co-incubated with 0.2X Enhancer for up to three days. Cells were imaged by confocal microscopy without a wash step.
CellBrite® Steady staining is more stable than CellMask™ Green for real-time live cell imaging. Live MCF7 cells were stained with 1X CellBrite™ Steady 488 with CellBrite® Steady Enhancer in complete culture medium at 37°C without a wash step.
Live HeLa cells stained with the lipophilic carbocyanine dyes Neuro-DiO, DiI, or DiD or CellBrite® Steady Membrane Dyes and Enhancer. Lipophilic carbocyanine dyes are rapidly internalized into intracellular compartments, so surface staining must be imaged within short time frames. CellBrite® Steady Dyes stain cell membranes evenly, and surface staining can be clearly imaged for 24 hours with the use of Enhancer.
CellBrite® Steady Membrane Staining Kits - Image 5 Comparison of live cell staining with CellBrite™ Steady Dyes with or without Enhancer. Live HeLa cells were stained for the indicated times. Over 24 hours, the dyes equilibrate between the plasma membrane and intracellular compartments. Co-incubating or post-treating with Enhancer masks intracellular staining, allowing selective imaging of cell boundaries. Cells were imaged by confocal microscopy without a wash step.
Excitation and emission spectra of CellBrite™ Steady Dyes in methanol.
Live HeLa cells co-stained with CellBrite™ Steady 488 (green) with Enhancer and NucSpot® Live 650 (magenta) for 45 minutes. Cells were imaged by confocal microscopy without a wash step.
HeLa cells co-stained with CellBrite® Steady 488 (green) and MitoView™ 633 mitochondrial stain (magenta).
CellBrite® Steady Membrane Staining Kits - Image 7 Long-term staining with CellBrite™ Steady Dyes and Enhancer. MCF7 cells stained with 1X CellBrite™ Steady 405 or CellBrite™ Steady 488 dye, co-incubated with 0.2X Enhancer for up to six days. Cells were imaged by confocal microscopy without a wash step.
Live MCF7 cells stained with CellBrite® Steady dyes for 30 minutes and imaged by confocal microscopy without a wash step.
Enriched EVs were stained with 0.5 uM CellBrite® Steady 650 (right). Specific staining was seen, compared with CellBrite® Steady 650 in buffer (left). EVs were detected on a Cytoflex LX flow cytometer in the APC channel.
Enriched EVs were stained with 0.5 uM CellBrite® Steady 550 (right). Specific staining was seen, compared with CellBrite® Steady 550 in buffer (left). EVs were detected on a Cytoflex LX flow cytometer in the PE channel.
Enriched EVs were stained with 0.5 uM CellBrite Steady® 488 (right). Specific staining was seen, compared with CellBrite® Steady 488 in buffer (left). EVs were detected on a Cytoflex LX flow cytometer in the FITC channel.
SEC-purified MCF-7-derived exosomes were stained with 0.02X CellBrite® Steady 405 (right). Specific staining was seen, compared with CellBrite® Steady 405 in buffer (left). EVs were detected on a Cytoflex LX flow cytometer in the Pacific Blue channel.
CellBrite® Steady Membrane Staining Kits - Image 10 Enriched EVs were stained with 0.5 uM CellBrite® Steady 405 (right). Specific staining was seen, compared with CellBrite® Steady 405 in buffer (left). EVs were detected on a Cytoflex LX flow cytometer in the Pacific Blue channel.

CellBrite™ Steady Membrane Staining Kits

CellBrite® Steady Membrane Staining Kits label live cell membranes with bright, even staining and improved surface retention for 24 hours or longer, outperforming rapidly internalised dyes.

CellBrite™ Steady Membrane Staining Kits

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Live cell surface membrane staining that stays sharp for 24hours or longer

CellBrite® Steady Membrane Staining Kits enable bright, even live cell membrane staining that remains useful for 24 hours or longer, helping maintain clear cell boundaries during extended imaging. Unlike many membrane dyes that rapidly internalise and lose surface definition, CellBrite® Steady Dyes retain membrane outlining over time. When a cleaner surface signal is required, the included CellBrite® Steady Enhancer can mask intracellular fluorescence to improve visualisation of cell outlines.

Key Features

  • Retained on the cell surface longer than many membrane dyes that internalise rapidly
  • Live cell membrane imaging for 24 hours or longer, depending on cell type and proliferation
  • Optional Enhancer reduces intracellular signal for clearer membrane boundaries
  • Rapid, even staining in complete culture medium
  • Broad colour range from blue to near IR, with STORM compatible options (550, 650, 685)
  • Low toxicity for continuous incubation in culture medium, with usage guidance for Enhancer

 

Kit Components

Kit sizes are based on a 200 µL labelling volume.

Kit Size Component Concentration / Format Volume
Trial Size (100 labelings) CellBrite® Steady Dye 1000X in DMSO 20 µL
CellBrite® Steady Enhancer 1000X in DMSO 20 µL
Full Size (500 labelings) CellBrite® Steady Dye 1000X in DMSO 100 µL
CellBrite® Steady Enhancer 1000X in DMSO 100 µL

 

CellBrite® Steady staining is more stable than CellMask™ Green for real-time live cell imaging. Live MCF7 cells were stained with 1X CellBrite™ Steady 488 with CellBrite® Steady Enhancer in complete culture medium at 37°C without a wash step.
CellBrite® Steady staining is more stable than CellMask™ Green for real-time live cell imaging. Live MCF7 cells were stained with 1X CellBrite™ Steady 488 with CellBrite® Steady Enhancer in complete culture medium at 37°C without a wash step. Cells were cultured for 72 hours after staining and imaged at the time points shown. For CellMask™ Green staining, cell were stained according to the manufacturer’s recommendations. Cells were stained with dye diluted 1:1000 in HBSS buffer for 10 minutes at 37°C, then rinsed in buffer, and then fresh culture medium was added. Cells were cultured for 72 hours and imaged at the time points shown.
Live HeLa cells stained with the lipophilic carbocyanine dyes Neuro-DiO, DiI, or DiD or CellBrite® Steady Membrane Dyes and Enhancer. Lipophilic carbocyanine dyes are rapidly internalized into intracellular compartments, so surface staining must be imaged within short time frames. CellBrite® Steady Dyes stain cell membranes evenly, and surface staining can be clearly imaged for 24 hours with the use of Enhancer.
Live HeLa cells stained with the lipophilic carbocyanine dyes Neuro-DiO, DiI, or DiD or CellBrite® Steady Membrane Dyes and Enhancer. Lipophilic carbocyanine dyes are rapidly internalized into intracellular compartments, so surface staining must be imaged within short time frames. CellBrite® Steady Dyes stain cell membranes evenly, and surface staining can be clearly imaged for 24 hours with the use of Enhancer.

Our customer feedback to us that:

“The dye has been performing perfectly fine in my hands—actually better than CellMask. It is brighter, gives clearer membrane definition, and the overall consistency is very good. The protocol is straightforward, and I haven’t faced any issues with background or toxicity.”

Recommended Use Cases

  • Live cell membrane outlining for confocal and widefield microscopy
  • Cellular uptake studies where membrane definition must remain clear over time
  • Multi colour imaging panels from 405 to near IR
  • Super resolution imaging by STORM using Steady 550, 650, or 685
  • Purified extracellular vesicle labelling, with guidance below for bead bound formats

Tips for Success

  • Washing after staining is optional for confocal imaging, but required for epifluorescence imaging
  • For visible range dyes (405, 488, 550), no wash confocal imaging may require brightfield or another marker for initial focusing
  • Enhancer may be toxic to some cell types at higher concentrations; consider adding after staining or using the lowest effective concentration
  • Enhancer masking tends to be more complete with far red Steady 650 and Steady 685

 

Important Notes

  • For live cell staining only. Not suitable for staining already fixed cells
  • Formaldehyde fixation retains signal initially, but signal can redistribute during storage; methanol and detergents are not tolerated
  • Enhancer does not tolerate fixation. If Enhancer treated cells are fixed, masking is lost and intracellular fluorescence returns
  • Not recommended for cell tracking, co culture, or transplantation, as dyes can transfer between cells
  • EV labelling: validated for purified EVs, but may bind non specifically to bead bound exosomes; for bead bound and purified EVs, consider ExoBrite™ True EV Membrane Stains


P/s: Biotium products are distributed only in Singapore and Thailand

Documents

Protocol:

CellBrite™ Steady Membrane Staining Kits protocol 

MSDS: 

CellBrite™ Steady Membrane Staining Kits SDS

Brochure: 

CellBrite™ Steady Membrane Staining Kits brochure 

FAQs:

CellBrite™ Steady Membrane Staining Kits FAQs

Variations

Product Ex/Em Laser line(s) (nm) Detection channel Trial size (100 labelings) Full size (500 labelings)
CellBrite® Steady 405 Membrane Staining Kit 406/428 405 DAPI 30105-T 30105
CellBrite® Steady 488 Membrane Staining Kit 505/529 488 FITC 30106-T 30106
CellBrite® Steady 550 Membrane Staining Kit 562/579 555 or 561 TRITC or Rhodamine 30107-T 30107
CellBrite® Steady 568 Membrane Staining Kit 569/586 555 or 561 Alexa Fluor® 568 30150-T 30150
CellBrite® Steady 594 Membrane Staining Kit 580/615 555, 561, or 594 Texas Red® 30142-T 30142
CellBrite® Steady 650 Membrane Staining Kit 656/676 640 Alexa Fluor® 647 30108-T 30108
CellBrite® Steady 685 Membrane Staining Kit 686/708 685 Alexa Fluor® 680 30109-T 30109
CellBrite® Steady 750 Membrane Staining Kit 755/789 730 Alexa Fluor® 750 30141-T 30141

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