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LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells - FAQs

LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells - FAQs

查看更多产品信息 LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells - FAQs (L3224)

7 个常见问题解答

我在 Countess II FL全自动细胞计数仪上能进行哪种荧光活性实验？

我们已经确认可在Countess II FL全自动细胞计数仪上使用下列试剂盒：

•LIVE/DEAD Viabilty/Cytoxicity试剂盒（货号L3224）含有钙黄绿素AM和溴乙啡锭二聚体
•ReadyProbes Cell Viability Imaging试剂盒，蓝/绿（货号R37609）
•ReadyProbes Cell Viability Imaging试剂盒，蓝/红（货号R37610）含有NucBlue Live/NucGreen Dead和NucBlue Live/propidium iodide

见此应用说明(https://www.thermofisher.com/content/dam/LifeTech/global/life-sciences/Laboratory%20Instruments/Files/0415/CO014723-Countess-II-FL-Viability-Appnote_FHR.pdf)中的详情。

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。

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.

I am doing a Live/Dead assay using Calcein, AM, for live cells and ethidium homodimer-1 for dead cells. Can I fix the cells after labeling and retain the staining?

This is not recommended. Neither Calcein nor ethidium homodimer-1 bind to any cellular components upon fixation. There is no guarantee that the dyes will be retained upon fixation or any subsequent wash steps. We recommend scoring for live and dead cells as soon as possible after staining.

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

How should I store the LIVE/DEAD Viability/Cytotoxicity Kit, for mammalian cells (Cat. No. L3224)?

We recommend storing the LIVE/DEAD Viability/Cytotoxicity Kit, for mammalian cells (Cat. No. L3224) in the freezer at -5 degrees C to -30 degrees C and protected from light.

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

What fluorescent viability assays can I use on the Countess II FL automated cell counter?

We have validated the following kits for use on the Countess II FL Automated Cell Counter:

LIVE/DEAD Viabilty/Cytoxicity Kit (Cat. No. L3224) containing calcein AM and ethidium homodimer-1
ReadyProbes Cell Viability Imaging Kit, Blue/Green (Cat. No. R37609)
ReadyProbes Cell Viability Imaging Kit, Blue/Red (Cat. No. R37610) containing NucBlue Live/NucGreen Dead and NucBlue Live/propidium iodide
See this Application Note for details - https://www.thermofisher.com/content/dam/LifeTech/global/life-sciences/Laboratory%20Instruments/Files/0415/CO014723-Countess-II-FL-Viability-Appnote_FHR.pdf.

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.