Comparison between 355 nm and 407 nm Laser Exciting Hochest 33342 in the Detection of Apoptosis_West China Medical Journal

Authors：

LI Xue ¹ , WU Yalan ¹ , HUANG Xin ² , HUANG Qiaorong ¹ ,  MENG Wentong. ¹

1. Labortory of Stem Cell Biology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P. R. China; 2. Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, Sichuan 610041, P. R. China;

Corresponding author：

MENG Wentong., Email: lixue-87@163.com

Keywords：

355 nm激光; 407 nm激光; 流式细胞仪; 凋亡; Hoechst 33342

DOI：

10.7507/1002-0179.20130274

Video：

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目的　比较使用流式细胞仪355 nm和407 nm激光器激发Hochest33342检测细胞凋亡。方法　通过ATO药物诱导急性早幼粒白血病细胞（NB4）及血清饥饿法诱导人肺癌细胞（NCl-H292）细胞凋亡，取24、48 h时间点收集细胞，进行Hoechst33342-碘化丙啶（PI）双染，分别在配置有两种激光器的流式细胞仪上检测细胞凋亡。结果　细胞经处理后24 h，355 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（28.20 ± 4.80）%；NCl-H292细胞凋亡率Hoechst33342+/PI-：（22.47 ± 2.78）%。407 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（25.10 ± 6.19）%。NCl-H292细胞凋亡率Hoechst33342+/PI-：20.47 ± 1.46%。处理后48 h，355 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（33.60 ± 3.75）%。NCl-H292细胞凋亡率Hoechst33342+/PI-：（26.77 ± 1.16）%。407 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（29.47 ± 2.33）%。NCl-H292细胞凋亡率Hoechst33342+/PI-：（31.47 ± 3.05）%。两种细胞处理后比处理前凋亡率明显升高，但355 nm激光器与407 nm激光器检测的凋亡结果差异不明显（P＞0.05）。结论　407 nm激光器激发Hoechst33342可检测细胞凋亡。

Citation： LI Xue,WU Yalan,HUANG Xin,HUANG Qiaorong,MENG Wentong.. Comparison between 355 nm and 407 nm Laser Exciting Hochest 33342 in the Detection of Apoptosis. West China Medical Journal, 2013, 28(6): 875-878. doi: 10.7507/1002-0179.20130274 Copy

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4.	徐晓雪, 邓君, 邹林樾, 等. 407nm激光流式细胞仪检测小鼠骨髓侧群细胞效果研究[J]. 继续医学教育, 2011, 25(11): 63-66.
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7.	van Engeland M, Nieland LJ, Ramaekers FC, et al. Annexin V-affinity assay: a review on an apoptosis detection system based on phosphatidylserine exposure[J]. Cytometry, 1998, 31(1): 1-9.
8.	Komoriya A, Packard BZ, Brown MJ, et al. Assessment of caspase activities in intact apoptotic thymocytes using cell-permeable fluorogenic caspase substrates[J]. J Exp Med, 2000, 191(11): 1819-1828.
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11.	王文静. 侧群细胞的研究进展[J]. 国际口腔医学杂志, 2011, 38(6): 665-669.
12.	Sanz MA, Grimwade D, Tallman MS, et al. Management of acute promyelocytic leukemia: recommendations from an expert panel on behalf of the European LeukemiaNet[J]. Blood, 2009, 113(9): 1875-1891.
13.	Shen ZX, Chen GQ, Ni JH, et al. Use of Arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia (APL): II. Clinical efficacy and pharmacokinetics in relapsed patients[J]. Blood, 1997, 89(9): 3354-3360.
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1. 　Wlodkowic D, Telford W, Skommer J, et al. Apoptosis and beyond: cytometry in studies of programmed cell death [J]. Methods Cell Biol, 2011, 103: 55-98.
2. 　Latt SA, Stetten G. Spectral studies on 33258 Hoechst and related bisbenzimidazole dyes useful for fluorescent detection of deoxyribonucleic acid synthesis[J]. J Histochem Cytochem, 1976, 24(1): 24-33.
3. 　Veremes I, Haanen C, Reutelingsperger C. Flow cytometry of apoptotic cell death[J]. J Immunol Methods, 2000, 243(1-2): 167-190.
4. 　徐晓雪, 邓君, 邹林樾, 等. 407nm激光流式细胞仪检测小鼠骨髓侧群细胞效果研究[J]. 继续医学教育, 2011, 25(11): 63-66.
5. 　Simpson C, Pearce DJ, Bonnet D, et al. Out of the blue: a comparison of Hoechst side population (SP) analysis of murine bone marrow using 325, 363 and 407 nm excitation sources[J]. J Immunol Methods, 2006, 310(1-2): 171-181.
6. 　Telford WG, Frolova EG. Discrimination of the hoechst side population in mouse bone marrow with violet and near-ultraviolet laser diodes[J]. Cytometry A, 2004, 57(1): 45-52.
7. 　van Engeland M, Nieland LJ, Ramaekers FC, et al. Annexin V-affinity assay: a review on an apoptosis detection system based on phosphatidylserine exposure[J]. Cytometry, 1998, 31(1): 1-9.
8. 　Komoriya A, Packard BZ, Brown MJ, et al. Assessment of caspase activities in intact apoptotic thymocytes using cell-permeable fluorogenic caspase substrates[J]. J Exp Med, 2000, 191(11): 1819-1828.
9. 　Mathur A, Hong Y, Kemp BK, et al. Evaluation of fluorescent dyes for the detection of mitochondrial membrane potential changes in cultured cardiomyocytes[J]. Cardiovasc Res, 2000, 46(1): 126-138.
10. 　Mocharla R, Mocharla H, Hodes ME. A novel, sensitive fluorometric staining technique for the detection of DNA in RNA preparations[J]. Nucleic Acids Res, 1987, 15(24): 10589.
11. 　王文静. 侧群细胞的研究进展[J]. 国际口腔医学杂志, 2011, 38(6): 665-669.
12. 　Sanz MA, Grimwade D, Tallman MS, et al. Management of acute promyelocytic leukemia: recommendations from an expert panel on behalf of the European LeukemiaNet[J]. Blood, 2009, 113(9): 1875-1891.
13. 　Shen ZX, Chen GQ, Ni JH, et al. Use of Arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia (APL): II. Clinical efficacy and pharmacokinetics in relapsed patients[J]. Blood, 1997, 89(9): 3354-3360.
14. 　Liu JW, Chandra D, Rudd MD, et al. Induction of prosurvival molecules by apoptotic stimuli: involvement of FOXO3a and ROS[J]. Oncogene, 2005, 24(12): 2020-2031.

West China Medical Journal

Comparison between 355 nm and 407 nm Laser Exciting Hochest 33342 in the Detection of Apoptosis

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