DiO'; DiOC18(3) (3,3'-Dioctadecyloxacarbocyanine Perchlorate)
DiO'; DiOC<sub>18</sub>(3) (3,3'-Dioctadecyloxacarbocyanine Perchlorate)
Citations & References (188)
Invitrogen™

DiO'; DiOC18(3) (3,3'-Dioctadecyloxacarbocyanine Perchlorate)

The green fluorescent, lipophilic carbocyanine DiOC18(3) is widely used as a lipophilic tracer. It is weakly fluorescent in water, butRead more
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Catalog NumberQuantity
D275100 mg
Catalog number D275
Price (USD)
-
Quantity:
100 mg

The green fluorescent, lipophilic carbocyanine DiOC18(3) is widely used as a lipophilic tracer. It is weakly fluorescent in water, but highly fluorescent and quite photostable when incorporated into membranes. It has an extremely high extinction coefficient and short excited-state lifetimes (∼1 nanosecond) in lipid environments. Once applied to cells, the dye diffuses laterally within the plasma membrane.

We recommend preparing stock solutions of lipophilic tracers in dimethyl formamide (DMF), dimethylsulfoxide (DMSO), or ethanol at 1 to 2.5 mg/mL. DMF is preferable to ethanol as a solvent for DiO.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
ColorGreen
Detection MethodFluorescence
Emission501 nm
Excitation Wavelength Range484 nm
For Use With (Equipment)Fluorescence Microscope
Quantity100 mg
Shipping ConditionRoom Temperature
Product TypeLiphophilic Tracer
SubCellular LocalizationCell Membranes, Lipids
Unit SizeEach
Contents & Storage
Store at room temperature and protect from light.

Frequently asked questions (FAQs)

I'm labeling live cells with Vybrant DiI or DiD lipophilic cyanine dyes. DiI gives a nice even membrane labeling, but DiD is more "spotty". What can be done?

This is expected. DiD (which is far-red fluorescent) is never as uniform as DiI (which is orange fluorescent). If uniformity is desired, try increasing the label time and concentration, but it still isn't likely to be as uniform as DiI. CellMask Deep Red Plasma Membrane stain is much more uniform and is about the same wavelength as DiD. However, if you intend to do cell tracking over days, CellMask stain has not been tried for that application.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I want to perform a cell fusion assay, where one cell line is labeled with one color and the other cell line with another color, and combine with a nucleic acid stain. What do you recommend?

A typical method is to label one cell line with orange fluorescent DiI C18 and the other cell line with green fluorescent DiO C18. These orange and green lipophilic cyanine dyes will stain the membranes of cells. Cells that fuse will then have both dyes, yielding a yellow color (when images are overlaid or cells are imaged in a dual-bandpass filter). These live cells can then be labeled with Hoechst 33342 (a cell-permeant blue DNA stain comparable in wavelength to DAPI), but only as an endpoint just before imaging (since DNA stains can interrupt DNA function).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I need to look at live cell morphology deformation over the course of a few hours. What sort of membrane dye would be useful for this?

Lipophilic cyanine dyes, such as DiI (Cat. No. D282), DiO (Cat. No. D275), DiD (Cat. No. D7757) or DiR (Cat. No. D12731), are commonly used. The longer the alkyl chain on the dye, the better the retention in lipophilic environments.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I stained my cells with a lipophilic cyanine dye, like DiI, but the signal was lost when I tried to follow up with antibody labeling. Why?

Since these dyes insert into lipid membranes, any disruption of the membranes leads to loss of the dye. This includes permeabilization with detergents like Triton X-100 or organic solvents like methanol. Permeabilization is necessary for intracellular antibody labeling, leading to loss of the dye. Instead, a reactive dye such as CFDA SE should be used to allow for covalent attachment to cellular components, thus providing for better retention upon fixation and permeabilization.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

How long does it take for lipophlic tracers to transport along the membrane? How much faster are the FAST lipophilic dyes?

The transport is fairly slow, around 6 mm/day in live tissue and slower in fixed tissue, so diffusion of lipophilic carbocyanine tracers from the point of their application to the terminus of a neuron can take several days to weeks The FAST DiO and DiI analogs (which have unsaturated alkyl tails) can improve transport rate by around 50%.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

View all DiO'; DiOC18(3) (3,3'-Dioctadecyloxacarbocyanine Perchlorate) FAQs

Citations & References (188)

Citations & References
Abstract
Subzonal organization of olfactory sensory neurons projecting to distinct glomeruli within the mouse olfactory bulb.
Authors:Levai O, Breer H, Strotmann J
Journal:J Comp Neurol
PubMed ID:12619077
'Olfactory sensory neurons located in the nasal neuroepithelium send their axons directly into the olfactory bulb, where they contact the dendrites of second-order neurons in specialized spherical structures called glomeruli; each sensory neuron projects to a single glomerulus. All neurons expressing the same odorant receptor gene are confined to distinct ... More
Direct membrane protein-DNA interactions required early in nuclear envelope assembly.
Authors:Ulbert S, Platani M, Boue S, Mattaj IW
Journal:J Cell Biol
PubMed ID:16717124
'Among the earliest events in postmitotic nuclear envelope (NE) assembly are the interactions between chromatin and the membranes that will fuse to form the NE. It has been proposed that interactions between integral NE proteins and chromatin proteins mediate initial membrane recruitment to chromatin. We show that several transmembrane NE ... More
Antigen-induced translocation of PKC-theta to membrane rafts is required for T cell activation.
Authors:Bi K, Tanaka Y, Coudronniere N, Sugie K, Hong S, van Stipdonk MJ, Altman A
Journal:Nat Immunol
PubMed ID:11376344
'Protein kinase C-theta (PKC-theta) is essential for mature T cell activation; however, the mechanism by which it is recruited to the TCR signaling machinery is unknown. Here we show that T cell stimulation by antibodies or peptide-major histocompatibility complex (MHC) induces translocation of PKC-theta to membrane lipid rafts, which localize ... More
Beta-very low density lipoprotein is sequestered in surface-connected tubules in mouse peritoneal macrophages.
Authors:Myers JN, Tabas I, Jones NL, Maxfield FR
Journal:J Cell Biol
PubMed ID:8253839
'beta-very low density lipoprotein (VLDL) is a large lipoprotein with multiple apoprotein E (apoE) molecules that bind to the LDL receptors on mouse macrophages. Even though they bind to the same receptor, the endocytic processing of beta-VLDL differs from low density lipoprotein (LDL). LDL is rapidly delivered to perinuclear lysosomes ... More
Regulation of C-cadherin function during activin induced morphogenesis of Xenopus animal caps.
Authors:Brieher WM, Gumbiner BM
Journal:J Cell Biol
PubMed ID:8034750
'Treatment of Xenopus animal pole tissue with activin results in the induction of mesodermal cell types and a dramatic elongation of the tissue. The morphogenetic movements involved in the elongation appear similar to those in normal gastrulation, which is driven by cell rearrangement and cell intercalations. We have used this ... More
View all DiO'; DiOC18(3) (3,3'-Dioctadecyloxacarbocyanine Perchlorate) citations