摘要
目的建立一种分析方法证实流式细胞术对细胞凋亡分析的准确性。方法在流式细胞仪上利用Sub G1法、TdT法、AnnexinⅤ法和API法4种不同的凋亡细胞分析方法,对羟基喜树碱(CAM)诱导的急性T淋巴细胞白血病细胞株(Jurkat细胞)进行流式细胞术分析并同时分选出不同时相的凋亡和非凋亡细胞,再经激光扫描共聚焦显微镜(LSCM)进行形态学分析。结果①流式细胞术分选后共聚焦显微镜分析技术(PSC)能同时、准确地鉴别不同细胞周期时相的凋亡细胞和非凋亡细胞。②PSC技术能对同一群细胞内的凋亡细胞和坏死细胞进行形态学鉴别。③PSC技术证实了流式细胞术在细胞凋亡研究中的准确性。结论PSC技术结合了流式细胞术和LSCM的优点,更客观、准确地分析凋亡细胞,同时对细胞周期特异性的细胞凋亡有独到的优势,发展了细胞凋亡的分析方法。
Objective To estahlish a new analysis method to identify apoptotic and non-apoptotic cells in flow cytometry. Methods In flow cytometry, by using 4 apoptotic cell analyzing methods including sub-G1 , TdT, Annexin-Ⅴ and API, Jurkat cells induced by camptothecin （CAM） were analyzed, and apoptotic and non-apoptotic cells were sorted out simultaneously. The morphological changes were observed under the laser scanning confocal microscopy. Results Post-sorting confocal （PSC） method could exactly distinguish apoptotic and non-apoptotic cells in the specific cell cycle phases at the same time. The PSC method could distinguish apoptotic from necrotic cells morphologically in the same group cells. PSC method revealed that the flow cytometry was accurate in the study on the apoptosis. Apoptotic and non-apoptotic cells discerned by morphological characteristics in confocal consistent with flow cytometry analysis. Conclusion PSC combines the merits of flow cytometry and confocal and can analyze apoptosis more objectively and accurately, especially the cell cycle specific apoptosis. More important, it develops the analysis method for apuptosis.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2005年第4期486-489,F0003,共5页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.39670265
No.39730270
No.39725027)
卫生部科研基金资助项目(2020106)
国家973计划肿瘤资助项目(G1998051212)
