Cell surface staining kits with fluorescent membrane dye plus enhancer for imaging live cell surface for several hours to days.

• Retained on cell surface, unlike other membrane dyes that are rapidly internalised
• Image live cell surface membranes for 24 h or longer
• Optional Enhancer masks intracellular signal for selective imaging of cell surface
• Rapid, even staining in complete cell culture medium
• Dye colours from blue to near-IR, with STORM-compatible options

CellBrite™ Steady Membrane Staining Kits allow fluorescence imaging of cell surface for up to several days in culture. Unlike other membrane/cell surface stains that are rapidly lost from the cell surface by endocytosis after labelling, CellBrite™ Steady Dyes equilibrate between intracellular compartments and the plasma membrane. Cells retain surface staining in addition to intracellular staining over the course of hours to days in culture. CellBrite™ Steady Enhancer is an optional reagent included in the kits that can be used to mask intracellular fluorescence of CellBrite™ Steady Dyes, for more selective visualisation of cell boundaries.

CellBrite™ Steady Dyes are available in colors from blue to near-infrared. CellBrite™ Steady 550 and 650 fluorophores are compatible with super-resolution imaging by STORM. Masking of intracellular fluorescence by CellBrite™ Steady Enhancer tends to show more complete masking of intracellular fluorescence for far-red CellBrite™ Steady 650 and CellBrite™ Steady 685 dyes.

Washing after staining is optional for imaging by confocal microscopy, but is required for imaging by epifluorescence. For dyes with fluorescence in the visible range (405, 488, and 550), staining may or may not be visible through the microscope eyepieces without washing, so you may need to use brightfield or another marker to focus on cells initially for no-wash confocal imaging.

CellBrite™ Steady Dyes have low toxicity and can be continuously incubated in cell culture medium. Enhancer may be toxic to some cell types, especially at higher concentrations. Enhancer may be incubated together with membrane dyes or added after staining to minimise any potential effect of Enhancer on cells. See the Product Protocol for details.

Membrane staining with CellBrite™ Steady Dyes is retained immediately after fixation with formaldehyde, but does not tolerate methanol or detergent. If cells are stored after formaldehyde fixation, over time the dyes will redistribute to stain cytoplasmic structures. Enhancer does not tolerate fixation; if Enhancer-treated cells are fixed after staining, masking will be lost and intracellular fluorescence will return.