CellVue® Cell Labeling Kits
CellVue® fluorescent imaging kits use proprietary
labeling technology to stably incorporate fluorescent
dyes containing long aliphatic hydrocarbon tails into
lipid membranes (1). They are useful for researchers
working in all aspects of science and technology where
fluorescently labeled cells and/or tissues are required.
The molecular structure of CellVue® dyes consist
of long aliphatic hydrocarbon tails linked to a polar
fluorescent chromophore. These extremely lipophilic
fluorescent dyes rapidly and stably integrate into
the phospholipid membrane of cells or other membrane-containing
bioparticles by non-covalent interactions. The dyes
are stably maintained within the lipid bilayer through
strong hydrophobic interactions and do not transfer
into the unstained membranes of adjacent cells. This
characteristic permits a labeled cell to be tracked
for extended periods of time. The rapid incorporation
of the dye also allows for immediate analysis of cell
functions without any waiting period. The pattern of
fluorescence staining within the membrane may vary
depending on the cell type. All of the dyes are designed
to be biocompatible at optimized concentrations.
The CellVue® dyes are provided in an easy to use
kit format comprised of a solution of the dye in ethanol
and a cell labeling diluent. The labeling diluent provided
with the kit is an iso-osmotic aqueous solution which
contains no physiologic salts or buffers, detergents,
or organic solvents, and is designed to maintain cell
viability while maximizing dye solubility and staining
efficiency (2,3). The simple membrane labeling protocol
provided with the kit is applicable to almost any type
of cell or organism.
The excitation and emission properties of CellVue® Burgundy and CellVue® NIR815 dyes are compatible with a range of commercially available near-infrared fluorescent plate readers, flow cytometers, in vitro and in vivo fluorescent imaging systems, and confocal microscopes.
Due to the aforementioned properties, the CellVue® dyes provide the researcher with valuable tools for a multitude of in vivo and in vitro cell studies using fluorescent membrane labels.
Examples of Imaging with CellVue® Labeled Membranes
Figure 1 - Image showing CellVue® NIR815 in the capillary region of the lungs of a nude mouse as imaged on the Pearl™ Imager immediately following an intravenous injection via the tail vein. CellVue® NIR815 cell stain is represented in green; 680 nm signals due to chlorophyll in the animal’s diet are represented in red; both fluorescent signals are overlaid on a white light image of the mouse.
Figure 2 - Two-color imaging with CellVue® Burgundy and CellVue® NIR815 using LI-COR Odyssey® Imaging System
Panels A-C: A monocyte cell line (U937) and a T-Cell line (SUPT1) were independently labeled with CellVue Burgundy (red) and CellVue NIR815 (green) according to the standard labeling protocol (1 x 107 cells/mL, 2 ?M dye for 2-5 min at 25°C). Labeled monocytes and T-cells (106 cells/spot) were deposited separately but with some overlap on a glass fiber pad and imaged using the Odyssey Infrared Imaging System. The images display the same sample pad viewed through different channels and indicate minimal signal overlap between the two dyes. Image A was taken using the 700nm emission filter and shows only monocytes, Image B was taken using the 800nm emission filter and shows only T-cells and Image C was taken using combined 700 and 800nm filters and shows both cell populations clearly visible. (Images courtesy of Simone van de Waarsenburg and Dr David Anderson, Burnet Institute, Melbourne, Australia.)
Panel D-F: Mouse lymphocytes were stained with either 10 ?M CellVue® Burgundy or CellVue® NIR815 for 5 min at 37°C then washed 5x with media according to the standard labeling protocol. They were then spotted on a slide, separately or together, and imaged using the 700 and 800nm channels of the Odyssey Infrared Imaging System. Image D shows the lymphocytes stained with CellVue Burgundy (Red), Image E shows the mixture of both CellVue® Burgundy and CellVue® NIR815 labeled cells and Image F shows the lymphocytes labeled with CellVue® NIR815 (Green). Note that since the CellVue® Burgundy labeled cells were 10x as concentrated as the CellVue® NIR815 labeled cells, the former appear as a larger population but again there is minimal color overlap between the two dyes. (Images courtesy of Dr. Edward Roy, University of Illinois.)
