Apoptosis Quantification Using Flow Cytometry

Annexin V/PI apoptosis-detection kit (Nanjing KeyGen Biotech Co., Ltd.) was used to evaluate the apoptosis according to the manufacturer’s protocol. The stained cells were immediately examined using an EPICS XL-MCL FACScan flow cytometer (Beckman Coulter, Inc., Brea, CA, USA) and analyzed using FlowJo 8.7.1 software (Ashland, OR). The results showed healthy viable cells in the lower left quadrant (Q4) on the scatter plot as (FITC-/PI-). The lower right quadrant (Q3) represented the early stage apoptotic cells as (FITC + /PI-). The upper right quadrant (Q2) represented necrotic cells and late stage apoptotic cells as (FITC + /PI +). Apoptotic rate = percentage of early stage apoptotic cells (Q3) + percentage of late stage apoptotic cells (Q2) [15 (link)].

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Ren L., Yang J., Meng X., Zhang J, & Zhang Y. (2022). The promotional effect of microRNA-103a-3p in cervical cancer cells by regulating the ubiquitin ligase FBXW7 function. Human Cell, 35(2), 472-485.

Publication 2022

Annexin V/PI apoptosis-detection kit EPICS XL-MCL FACScan flow cytometer

Annexin v Apoptotic Cells Fitc Necrotic

Corresponding Organization : First Affiliated Hospital of Henan University of Science and Technology

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Apoptosis rate (percentage of early stage apoptotic cells + percentage of late stage apoptotic cells)

control variables

Staining with Annexin V/PI apoptosis-detection kit (Nanjing KeyGen Biotech Co., Ltd.)
Using an EPICS XL-MCL FACScan flow cytometer (Beckman Coulter, Inc., Brea, CA, USA) to examine the stained cells
Analyzing the results using FlowJo 8.7.1 software (Ashland, OR)

controls

Positive control: Not specified
Negative control: Not specified

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