Trypan Blue Stain (0.4%) - FAQs

很遗憾，这个问题没有确切的答案。非细胞透过染料的吸收速率取决于细胞类型、健康状态、营养物、吞食活力等。  

台盼蓝染料接触光线后可能会降解，这些污染物可能会促进形成沉淀。经过冷藏或冷冻后，台盼蓝也可能形成橙/红纤维状聚合物。 

台盼蓝是一种在活性群体中评估死细胞数量的细胞非通透性的染料。其主要应用原理是，它属于一种带电荷的染料，除非细胞膜损伤，否则无法进入细胞内。活（活性）细胞排斥该染料，但是死（无活性）细胞或者细胞膜损伤的细胞会染上蓝色。

台盼蓝会结合到血清蛋白和细胞蛋白，这会导致高的背景染色。如果背景太暗，计数前细胞应离心下来并在非蛋白培养基、缓冲液或普通生理盐水中重悬。

这取决于细胞使用的荧光染色的类型。台盼蓝是一个细胞膜非通透性的染料且可以淬灭荧光。其可以淬灭活细胞表面的荧光染色或死细胞内部荧光染色。

台盼蓝会染色细胞膜损伤的细胞，不能区分凋亡或坏死引起的细胞膜损伤。

有，您可以通过此链接(https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/cell-culture-protocols/trypan-blue-exclusion.html)找到方案。

我们推荐向1 mL细胞加入0.1 mL台盼蓝储液（终浓度为0.04% w/v）。根据样品和仪器，终浓度范围从0.04%到0.2%（w/v）。 

When counting cells that have been mixed with Trypan Blue, the Countess accounts for this 1:1 dilution and multiplies the count by 2.
When using the Countess 3 Standard Slide, the Trypan Blue correction box needs to be unchecked, since a 1:1 dilution was not made with Trypan Blue.

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

The Trypan Blue Stain (0.4%) can be warmed up to 37 degrees C numerous times. The entire 1 mL volume of the stain should be warmed up to ensure that you are getting the entire amount at the proper concentration of 0.4%; if you took out an aliquot to warm up, and left behind some precipitate, the aliquot that is warmed up may not contain the complete amount of dye and the final concentration may be less than 0.4%.

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

The shelf life of the product is 36 months from the date of production.

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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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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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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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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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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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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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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)

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

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.