Trypan Blue Solution, 0.4% - FAQs

View additional product information for Trypan Blue Solution, 0.4% - FAQs (15250061)

11 product FAQs found

How do I quench any fluorescence outside of my live cells and leave my intracellular staining intact?

There are a number of reagents used to quench fluorescence on the outside of live cells or in the media (background fluorescence). Our Backdrop Background Suppressor reagents are a great option to quench blue, green, and red fluorescent dyes, various media components and fluorescent proteins. Another commonly used reagent is the cell impermeant colorimetric dye, Trypan blue.

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How is Gibco Trypan Blue Solution, 0.4% (Cat. No. 15250061) sterilized?

Trypan Blue is filter sterilized through a 0.22 µm filter.

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What is Trypan Blue Solution, 0.4% dissolved in?

It is a 0.85% saline solution.

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How long can cells be exposed to trypan blue before all cells are stained with the dye?

Unfortunately, there is no definitive answer to this question. The rate at which the cell-impermeant dye is absorbed depends on the cell type, their state of health, nourishment, engulfment activity, etc.

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What causes trypan blue to form a precipitate?

Exposure to light may degrade the dye and these contaminants may promote precipitation. Trypan blue can also form orange/red fibrous aggregates if exposed to refrigeration or freezing temperatures.

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How does trypan blue solution stain viable cells?

Trypan blue is a cell impermeant stain used to estimate the number of dead cells in a viable population. Its utility is based on the fact that it is a charged dye and does not enter cells unless the membrane is compromised. Live (viable) cells exclude the dye but dead (non-viable) cells or cells with a compromised membrane are stained an intense blue.

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Prior to staining with trypan blue solution, do the cells need to be washed to remove the culture medium?

Trypan blue will bind to serum proteins as well as cellular proteins, which may result a high level of background staining. If the background is too dark, cells should be pelleted and resuspended in protein-free medium, buffer, or normal saline prior to counting.

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Can I combine trypan blue staining with fluorescence staining?

It depends on the type of fluorescent stain used on the cells. Trypan blue is a cell-impermeant chromophore that can quench fluorescence. It may quench fluorescent staining on the surface of live cells or internal fluorescent staining in dead cells.

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Can trypan blue solution be used to differentiate between apoptotic and necrotic cells?

Trypan blue will stain cells that have a compromised membrane. It cannot differentiate between compromised membranes caused by apoptosis or necrosis.

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Do you have a protocol for cell counting via trypan blue exclusion?

Yes, you can find the protocol at the following link (https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/cell-culture-protocols/trypan-blue-exclusion.html)

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How much Trypan Blue solution should I add to my cell suspension before counting?

We recommend adding 0.1 mL of 0.4% Trypan Blue stock solution to 0.1 mL of cells. However, depending upon the sample and instrumentation, the ratio of 0.4% Trypan Blue solution to the volume of cells can be varied from 1: 10 to 1:1 (v/v) so that the final concentration of Trypan Blue is 0.04% - 0.2%.

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