LIVE/DEAD™ Cell Imaging Kit (488/570) - FAQs

View additional product information for LIVE/DEAD™ Cell Imaging Kit (488/570) - FAQs (R37601)

11 product FAQs found

我如何为LIVE/DEAD细胞活力实验准备死细胞对照？

有两种简单的方法。一种是通过60°C放置20分钟热灭活细胞。第二种是将细胞放入70%乙醇。乙醇固定的细胞可以在冰箱中无限期储存直到使用，可达好几年。

1.离心细胞、使细胞沉淀并去除上清
2.固定细胞：在15mL含有细胞团的试管中加入10 mL70%冰乙醇，先开始逐滴加入后震荡，混合均匀。
3.在冰箱中储存直到使用。
4.快使用时，水洗两次且在选择的缓冲液中重悬。

我希望我的细胞活力实验持续好几天。我能使用你们的LIVE/DEAD成像试剂盒做这个实验么？

不可以，不可能测试相同群体细胞的活性超过几小时；您可以使用重复的样品。DNA结合的染料对细胞有毒；染色后细胞应尽快成像。钙黄绿素AM不能停留在细胞中，根据不同类型的细胞，在几分钟到若干小时内，其可能会被细胞主动地排出。钙黄绿素AM对细胞无毒，因此其可以重复添加到相同的样品中。您可以使用alamarBlue试剂或PrestoBlue试剂进行相同样品超过若干天的细胞增殖检测，因为这些染料对细胞是无毒性的。

I need to use a dead cell control for my viability assay. Do you have a protocol for killing cells for this?

Heat killing is commonly used. Place your cells in a tube in buffer and heat at 60oC for 20 minutes. You can also kill your cells by fixing them with ice cold 70% ethanol for 15 minutes. The ethanol-killed cells can then be stored at -20oC until needed, at which point you wash out the ethanol and replace with buffer.

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

Can I use the LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601) with fixed cells?

The LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601) is used for identifying live and dead cells using fluorescence-based staining. It cannot be used with fixed cells.

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

What is the composition of the Live Green (Component A) of the LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601)?

The formulation of the solution as well as the amount and concentration of Calcein, AM provided in the LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601) is proprietary information.

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

Is the LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601) only a one-time use or is there a way to aliquot and freeze it down?

Once the Live Green (Component A) and Dead Red (Component B) are mixed, this solution should be used immediately for one-time use and should not be stored.

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

What is the nuclear stain in the LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601)?

The nuclear stain in the LIVE/DEAD Cell Imaging Kit (488/570) is BOBO-3. The amount, concentration, and formulation is proprietary.

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

What cell density do you recommend using for the LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601)?

As a general guideline, cell density should be within the range of 1 x 10^6 cells/mL of staining solution. Higher cell densities may result in poor staining (low signal) and lower cell densities may result in over-staining.

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

Could Ca++ or Mg++ ions interfere with staining using LIVE/DEAD Cell Imaging Kit (488/570) (Cat. No. R37601)?

No, Ca++ or Mg++ ions do not interfere with staining using LIVE/DEAD Cell Imaging Kit (488/570) unless when used at high concentrations; salts at high concentrations can promote precipitation of various dyes.

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

How do I prepare dead cell controls for LIVE/DEAD cell viability assays?

There are two easy options. One is to heat-inactivate the cells by placing at 60 degrees C for 20 minutes. The second is to subject the cells to 70% ethanol. Alcohol-fixed cells can be stored indefinitely in the freezer until use, potentially up to several years.

Centrifuge cells, pellet, and remove supernatant.
Fix cells: Add 10 mL ice cold 70% ETOH to a 15 mL tube containing the cell pellet, adding dropwise at first while vortexing, mix well.
Store in freezer until use.
When ready to use, wash twice and resuspend in buffer of choice.

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

I wish to assay the viability of my cells over several days. Can I use any of your LIVE/DEAD imaging kits to do this?

No, it is not possible to assay the viability of the same population of cells longer than a few hours; you will need to use replicate samples. DNA binding dyes are toxic to cells; stained cells should be imaged as soon as possible after staining. Calcein AM is not retained in cells and may be actively effluxed out in the range from minutes to several hours, dependent upon the cell type. Calcein AM is not toxic to cells, so it can be added repeatedly to the same samples. You can assay the proliferation of the same sample of cells over several days using alamarBlue reagent or PrestoBlue reagent, as these dyes are non-toxic to cells.

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