CellEvent™ Caspase-3/7 Green Detection Reagent, 1 x 100 μL Vial - FAQs

我们已经证实CellEvent Caspase 3/7 Green Detection Reagent（货号C10423）结合 SYTOX Red Dead Cell Stain （货号S34859）可用于Countess II FL全自动细胞计数仪上从死亡细胞中鉴别凋亡细胞。更多详情，请见应用说明（https://www.thermofisher.com/content/dam/LifeTech/global/life-sciences/Laboratory%20Instruments/Files/0615/CO210384-CountessIIFL-Apoptosis-AppNote.pdf)。 

可以，在3.7%甲醛中固定15分钟后，CellEvent信号将会保留。 

可以在同种样品中将PrestoBlue试剂（用于细胞增殖）和CellEvent Caspase 3/7 Green 检测试剂（用于细胞凋亡检测）结合。

•可测定来自单个细胞的数据。
•可从大量细胞中获得数据，产生细胞群的丰富统计学分析结果。
•由于可测定单个细胞，能够揭示种群的异质性。
•支持多重检测，可鉴定小型亚群。
•可以快速分析数以千计的细胞。
•非常适合血液样本和其他悬浮细胞。
•数据获取后，可以多次重复分析。
•流式细胞仪文件（FCS）可以归档。

可进行多种应用，包括免疫分型、细胞周期分析、凋亡检测（如膜联蛋白V染色检测实验）、CellEvent Caspase-3/7检测、TUNEL检测、细胞活性检测、增殖检测（如CellTrace 检测和Click-iT EdU检测）、MitoProbe检测法测定线粒体电势、利用计数微球进行细胞计数。

Yes, on both accounts. We have demonstrated that it works for microplates, and we have multiplexed the dye with PrestoBlue Cell Viability Reagent. Some microplates may be less sensitive than microscopic analysis, so sensitivity may not be the same for the CellEvent reagent.

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

It has been done, but we don‘t recommend it. Both healthy cells and apoptotic cells possess phosphatidylserine on the cell surface, which can be detected with Annexin V, but apoptotic cells have significantly more of it. You can easily tell the difference between these two populations with flow cytometry, because flow cytometers are more sensitive and have a higher throughput. But with a microscope, you cannot always tell the difference, especially for adherent cells. Instead, for microscopy, we recommend a different technique, such as detecting caspases with CellEvent Caspase Detection Reagents.

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

We have tested Caspase 3/7 Green Detection Reagents (Cat. Nos. C10432, C10433, C10434, C10423, C10723, R37111) for up to 48 hours and observed no toxicity or stability issues with the reagent. If the samples are fixed at the end of the time course the signal will be retained for days. For live cells, apoptotic cells will round up and float away before the loss of fluorescence signal from CellEvent.

The following reference reports a visible signal for up to 72 hrs:

Sambi M, Samuel V, Qorri B, Haq S, Burov SV, Markvicheva E, Harless W, Szewczuk MR. A Triple Combination of Metformin, Acetylsalicylic Acid, and Oseltamivir Phosphate Impacts Tumour Spheroid Viability and Upends Chemoresistance in Triple-Negative Breast Cancer. Drug Des Devel Ther. 2020 May 25;14:1995-2019. doi: 10.2147/DDDT.S242514. PMID: 32546966; PMCID: PMC7260544.

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

No, it must be applied to live cell populations.

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

Yes, you may use SYTOX AADvanced Dead Cell Stain that is included in the CellEvent Caspase-3/7 Green Ready Flow Reagent (Cat. No. R37167) and in the CellEvent Caspase-3/7 Green Flow Cytometry Assay Kit (Cat. No. C10427, C10740). You may also try using other nucleic acid stains in the red or far red channels for staining cells.

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

Yes. You can apply surface labels or other reagents. We recommend applying the surface labels or other reagents first.

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

The CellEvent Caspase-3/7 Green Detection Reagent is not covalently attached to any cellular component and may be lost upon fixation. For flow cytometry applications, after applying the reagent, you should examine without washing or fixation. For imaging applications, the CellEvent product signal may be retained after fixation in 3.7% formaldehdye for 15 min.

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

We have validated CellEvent Caspase 3/7 Green Detection Reagent (Cat. No. C10423) in combination with SYTOX Red Dead Cell Stain (Cat. No. S34859) for discriminating apoptotic cells from dead cells using the Countess II FL Automated Cell Counter. For further details, please see this Application Note (http://www.thermofisher.com/content/dam/LifeTech/global/life-sciences/Laboratory%20Instruments/Files/0615/CO210384-CountessIIFL-Apoptosis-AppNote.pdf).

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

For imaging applications, we recommend fixing with 3.7% formaldehyde at room temperature for 15 mins. After fixation, cells can be imaged for up to 24 hrs, but the signal will gradually degrade over time. For flow cytometry applications, fixation is not recommended.

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

It is possible to combine PrestoBlue reagent (for cell proliferation) with the CellEvent Caspase 3/7 Green Detection Reagent (for apoptosis detection) in the same sample.

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

-Measures data from single cells.
-Data are obtained for a large number of cells, generating a rich statistical analysis of cell populations.
-Because single cells are measured, it will reveal heterogeneity within a population.
-With the ability to multiplex, small sub-populations can be identified.
-Thousands of cells can be analyzed rapidly.
-It is ideally suited for blood samples and other cells in suspension.
-Data can be re-analyzed multiple times after acquisition.
-Flow cytometry files (FCS) can be archived.

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

There are several applications, some of which include immunophenotyping, cell cycle analysis, apoptosis assays such as annexin V staining, CellEvent Caspase-3/7 assay, and TUNEL assay, cell viability, proliferation assays such as CellTrace assay and Click-iT EdU assay, measurements of mitochondrial potential with MitoProbe assays, and cell counting using counting beads.

Find additional tips, troubleshooting help, and resources within our Flow Cytometry Support Center.