Cell Function FAQs

No, the concentration of calcium ions in binding buffer is determined after experimental exploration, and PBS containing calcium ions may not have the effect of binding buffer.

It is not advised to use tissue samples for Annexin V experiments. On the one hand, in the process of preparing tissue samples into single-cell suspension, the apoptosis rate caused by the experimental operation may be higher than that of the sample itself. Moreover, the experimental operation is unstable, and the actual apoptosis rate of the sample itself cannot be determined by the final experiment. On the other hand, tissue samples have multiple cell types, each with different autofluorescence, and the results cannot be analyzed.

The FITC is 490nm/530nm and the 7AAD is 546nm/647nm (which can be excited with 488nm). Generally, it can be detected by FITC channel and PerCP channel.

Untransfected cells with obvious apoptosis were stained with 7-AAD and APC, respectively, as single positive control 1 and single positive control 2. The samples transfected with green fluorescence were directly put on the machine as single-positive control 3.

The cells in the control group can be induced apoptosis by the following two methods: ① Since different cells are affected by DMSO to different degrees, it is recommended to explore the action time or volume fraction of DMSO through pre-experiment. The action time can be explored according to the following volume fraction (10%) first, and the microscopic examination is conducted every 30 minutes. ② The cells were bathed in a water bath at 50~60℃ for 5~10min. When the cell morphology changes or a large number of floating, can be considered to produce apoptotic cell population.

Because during the culture process, apoptotic cells do not stick firmly to the flask, and a large number of them float in the culture medium. For the accuracy of the results, it is usually recommended that the suspended cells be collected and tested together with the cells digested by trypsin when detecting apoptosis.

We haven't tested it. Because single-celled diatoms have cell walls, it is recommended that customers do pre-experiments to explore the dyeing time and conditions.

After the cells were suspended by PBS, anhydrous ethanol pre-cooled by -20°C was added drop by drop, and the cells were dispersed into single cells by shaking while adding. The fixed time must be at least 1 hour.

In theory, placing it in a -20℃ refrigerator for 2-3 days after fixing has little impact on the results.

If the sample does not contain fluorescence, the red, green and blue fluorescence cell cycle kit all can be used, and the red fluorescence kit (E-CK-A351) is more commonly used. If the sample has green fluorescence, use the cell cycle blue fluorescence kit (E-CK-A353) without adjustment compensation. If the sample has red fluorescence, it is recommended to use cell cycle blue fluorescence kit (E-CK-A353) and cell cycle green fluorescence kit (E-CK-A352). If the sample has red and green fluorescence, the cell cycle blue fluorescence kit (E-CK-A353) is recommended. Before selecting E-CK-A353, check whether the flow cytometer has a DAPI channel.

The fluorescent dyes used are not the same, so the flow cytometry detection channel requirements are different from the applicable samples. The detection channel of red fluorescence is PI,PerCP and PE, the detection channel of green fluorescence is FITC, and the detection channel of blue fluorescence is DAPI.