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Image-IT™ DEAD Green™ Viability Stain - FAQs

查看更多产品信息 Image-IT™ DEAD Green™ Viability Stain - FAQs (I10291)

3 个常见问题解答

哪一种细胞活力试剂盒兼容固定？

•用于流式细胞仪分析的LIVE/DEAD可固定试剂盒适用于固定。这些试剂盒使用胺基反应细胞-非渗透染料对活细胞细胞表面和死细胞胞质染色——活细胞暗淡死细胞明亮。因为染料共价结合到细胞，固定后保留。不幸的是，此方法对基于成像的实验效果不好，由于所有细胞都被染色，很难通过显微镜从暗淡的活细胞中分辨出明亮的死细胞。

•Image-IT DEAD绿色活性染料（货号I10291），用于成像和高通量扫描（HCS）分析的，是一种活细胞非透过型DNA结合染料，固定且透过后可保留长达48小时。

•我们同时推出用于成像分析的LIVE/DEAD Reduced Biohazard Cell Viability试剂盒（货号L7013），其含有两种DNA结合染料，SYTO 10和Dead Red，可以有效染色且通过4%戊二醛固定后快速分析。

注：总的来说，DNA结合的染料和钙黄绿素AM不适用于固定，由于这些染料不能共价地结合到细胞组分上，因此会在固定后缓慢扩散出细胞，逐渐地将所有细胞都染色。

Do you offer a cell viability stain compatible with fixation?

Our Image-IT DEAD Green Viability Stain (Cat. No. I10291) is amenable to fixation and permeabilization after the application of the stain. It is important to use fixatives free of alcohol or other organic solvents.

Find additional tips, troubleshooting help, and resources within our Cell Viability, Proliferation, Cryopreservation, and Apoptosis Support Center.

Which cell viability kits are compatible with fixation?

The LIVE/DEAD Fixable kits for flow cytometry analysis are compatible with fixation. These kits use amine-reactive cell-impermeant dyes that stain the cell surface of live cells and also the cytosol of dead cells-live cells are dim and dead cells are bright. Since the dye is covalently bound to the cells, it will be retained after fixation. Unfortunately, this method does not work well for imaging-based assays, as all cells are stained and it is difficult to distinguish bright dead cells from dim live cells with a microscope. Ethidium monoazide (EMA; Cat No. E1374) is a cell impermeant nucleic acid stain that can be applied to live cultures and stains only dead cells. After incubation and washing away unbound dye, the cells can be exposed to light to photoactivate EMA to crosslink to dead cell DNA. After crosslinking to dead cell DNA, the samples may be fixed and permeabilized. Image-IT DEAD Green Viability Stain (Cat. No. I10291) for imaging and high-content screening (HCS) analysis is a live-cell impermeant DNA binding dye that is compatible with fixation and permeabilization with good retention up to 48 hours. We also have a LIVE/DEAD Reduced Biohazard Cell Viability Kit (Cat. No. L7013) for imaging and flow analysis that contains two DNA binding dyes, SYTO 10 and Dead Red, that are sufficiently retained to be analyzed soon after 4% glutaraldehyde fixation.
Note: In general, DNA-binding dyes and calcein AM are not compatible with fixation, as these dyes are not covalently bound to components of the cell and will thus slowly diffuse out of cells after fixation, gradually staining all cells as dead.

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