Annexin V experimental control setting

Annexin V experimental control setting

Annexin V is a common method to detect the apoptosis of cell, the experimental results are generally measured by flow cytometry. In order to ensure the accuracy of experimental results and facilitate subsequent data analysis, different control groups are needed.

There is no autofluorescence in the sample

There is autofluorescence in the sample

Blank: adjust the threshold and voltage.

Biological control: exclud the influence of experimental operation on experimental results.

Single positive: adjust compensation, auxiliary voltage regulation.

Attention：

* Obviously apoptotic cells must be used in the single positive control group because adequate positive signal is required for compensation adjustment

* There is autofluorescence in the sample, it is necessary to set up three tubes of single positive control to ensure that each tube has only one kind of fluorescence

Tips:

Adherent cells:

* Cells floating in the cell culture medium need to be collected

* Avoid cell damage caused by manual operation, for example, the time of trypsin digestion should be appropriate, and the operation should be gentle when digesting adherent cells

* Try not to use EDTA-containing trypsin, because EDTA affects the binding of Annexin V with PS. If the trypsin containing EDTA must be used, the cells should be washed clean after the termination of digestion to remove EDTA as far as possible

* Cells that are difficult to digest can be digested in batches, to avoid the first digested cells are over-digested causing false positive results