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Invitrogen™ Vybrant™ DiI Cell-Labeling Solution

Catalog No. V22885
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V22885 1 mL
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Catalog No. V22885 Supplier Invitrogen™ Supplier No. V22885
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Description

Lipophilic membrane stain that diffuses laterally to stain the entire cell

DiI is weakly fluorescent until incorporated into membranes. This orange-red-fluorescent dye, which is spectrally similar to tetramethylrhodamine, is often used as a long-term tracer for neuronal and other cells. DiI is also available as a solid (D-282), as a paste (N-22880) or as crystals (D-3911).

Cell Analysis, Cell Proliferation, Cell Tracing & Tracking, Cell Viability, Proliferation & Function, General Cell Tracing, Microbial Tracking, Neuronal Tracing

Order Info

Shipping Condition: Room Temperature

Specifications

Color Red-Orange
Content And Storage Store at room temperature and protect from light.
For Use With (Equipment) Fluorescence Microscope, Flow Cytometer
Quantity 1 mL
Detection Method Fluorescence
Product Line Vybrant
Shipping Condition Room Temperature
Label Type Fluorescent Dye
Product Type Cell Labeling
SubCellular Localization Cell Membranes, Lipids

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.
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).
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.

How do I know which tracer to choose for my experiment?

Factors to consider are size of tracer, method of delivery (injection, direct application to tissue, etc.), and if the tracer needs to be fixable. Here are some links to details about the various classes of neuronal tracers we offer and how to choose between them:

Neuronal Tracing (https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-tracing-tracking-and-morphology/neuronal-tracing.html)
Choosing a Tracer (https://www.thermofisher.com/us/en/home/references/molecular-probes-the-handbook/fluorescent-tracers-of-cell-morphology-and-fluid-flow/choosing-a-tracer.html)
Imaging Analysis (http://assets.thermofisher.com/TFS-Assets/BID/Reference-Materials/bioprobes-50-journal.pdf)

I want to track my cells with a nucleic acid stain, like DAPI or Hoechst dye. Do you recommend this?

This is not recommended. When these stains bind to DNA and RNA, they may affect the normal function of the nucleic acids, disrupting transcription, as well as replication. Other reagents, such as CellTracker dyes or Qtracker reagents are more optimized for tracking without disrupting normal activity. If a nuclear label is still desired, though, and the cells are mammalian and non-hematopoietic, CellLight nuclear reagents can transiently transfect cells to express GFP or RFP on a nuclear-expressing protein for up to several days without affecting function.
I want to track my cells over time, and you have a lot of options to choose from. How do I pick the right one?

Please see this Web link (http://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-tracing-tracking-and-morphology/cell-tracking.html) to help you choose the right option for your application. Start by planning how long you want to track your cells, then consider the mechanism of binding. Calcein dyes are very uniform in label and are good for short-term cell migration, but may be rapidly effluxed from some cell types. Lipophilic cyanine dyes, such as DiI, DiO, and similar dyes label cell membranes, don’t disrupt function, and can last longer, but have the potential to cross to other cells if membranes fuse. They are also lost upon permeabilization. CellTracker dyes are better for longer-term labeling, as they possess a mildly reactive chloromethyl moiety that allows covalent binding to cellular components. CFDA SE also covalently binds to cellular components. With all the reagents, their retention within cells is dependent upon the rate of cell division and the inherent properties of the cell (active efflux, membrane and protein turnover rates, etc.) and reagents that allow for covalent attachment exhibit longer retention than those that do not.

The longest-lasting and brightest options are the Qtracker reagents, which are taken up through endocytosis. These are so bright individual quantum dots can be detected, and are also robust enough to survive not only fixation and permeabilization, but even the heat and solvents used in paraffin processing.
I want to label the plasma membrane of my cells, but there are several dyes to choose from. Which one should I use?

For live-cell imaging, the CellVue and CellMask Plasma Membrane Stains are the most uniform and the slowest to be endocytosed. However, they are not the best choice if you wish to fix and permeabilize your cells, such as for antibody labeling. Wheat germ agglutinin (WGA) conjugates are also able to label live cells, or can label already formaldehyde-fixed cells. They can survive subsequent permeabilization with detergents, such as Triton X-100. If cells are already permeabilized, WGA will label internal structures as well. Thus, only an antibody against a plasma membrane protein can be used if cells are already permeabilized. Lipophilic cyanine dyes, such as DiI, will label all cell membranes in live cells, not just plasma membranes, if left on live cells for extended periods. Following page will help you choose (http://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-structure/plasma-membrane.html).

For Research Use Only. Not for use in diagnostic procedures.