Calcein—AM和Annexin V在流式细胞术检测早期细胞凋亡的敏感性比较
王佳, 冯永东, 陶德定, 李小兰, 肖薇, 刘双又, 余从年, 龚建平
Abstract
王佳, 冯永东, 陶德定, 李小兰, 肖薇, 刘双又, 余从年, 龚建平
Abstract
Objective The current study was designed to compare the sensitivities of Calcein-AM and Annexin V-FITC for early detection of apoptosis in living cells by flow cytometry and to establish a new method for early detection of apoptosis. Methods The Molt-4 cell line treated with camptothecin (CPT) or ultraviolet (UV) were labeled with Calcein-AM and FITC-conjugated Annexin V respectively, then the apoptosis rate were analyzed by flow cytometry and the performance of Calcein-AM and Annexin V for early detection of apoptosis were compared. Results Our results showed that both probes allowed the detection of apoptotic cells. However, Calcein-AM was more sensitive than Annexin V. We could detect the apoptosis induced by CPT in two hours after inducing with Calcein-AM and in three hours after inducing with Annexin V. The apoptosis induced by UV could be detected in one hours after inducing with Calcein-AM but two hours after inducing with Annexin V. Conclusion Annexin V is less sensitive than Calcein-AM for detection of early apoptosis, and the new method for detection of early apoptosis with Calcein-Am is stable, reliable, sensitive and low-cost.
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Objective The current study was designed to compare the sensitivities of Calcein-AM and Annexin V-FITC for early detection of apoptosis in living cells by flow cytometry and to establish a new method for early detection of apoptosis. Methods The Molt-4 cell line treated with camptothecin (CPT) or ultraviolet (UV) were labeled with Calcein-AM and FITC-conjugated Annexin V respectively, then the apoptosis rate were analyzed by flow cytometry and the performance of Calcein-AM and Annexin V for early detection of apoptosis were compared. Results Our results showed that both probes allowed the detection of apoptotic cells. However, Calcein-AM was more sensitive than Annexin V. We could detect the apoptosis induced by CPT in two hours after inducing with Calcein-AM and in three hours after inducing with Annexin V. The apoptosis induced by UV could be detected in one hours after inducing with Calcein-AM but two hours after inducing with Annexin V. Conclusion Annexin V is less sensitive than Calcein-AM for detection of early apoptosis, and the new method for detection of early apoptosis with Calcein-Am is stable, reliable, sensitive and low-cost.
Key concepts: Calcein, Annexin, Flow cytometry, Apoptosis, Molecular biology, Chemistry, Annexin A5, Cell biology