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; 流式細胞儀; 凋亡; Hoechst 33342

DOI：

10.7507/1002-0179.20130274

Video：

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Abstract Full text Figures/Tables Video References Cited by

目的　比較使用流式細胞儀355 nm和407 nm激光器激發Hochest33342檢測細胞凋亡。方法　通過ATO藥物誘導急性早幼粒白血病細胞（NB4）及血清饑餓法誘導人肺癌細胞（NCl-H292）細胞凋亡，取24、48 h時間點收集細胞，進行Hoechst33342-碘化丙啶（PI）雙染，分別在配置有兩種激光器的流式細胞儀上檢測細胞凋亡。結果　細胞經處理后24 h，355 nm激光器檢測NB4細胞凋亡率Hoechst33342+/PI-：（28.20 ± 4.80）%；NCl-H292細胞凋亡率Hoechst33342+/PI-：（22.47 ± 2.78）%。407 nm激光器檢測NB4細胞凋亡率Hoechst33342+/PI-：（25.10 ± 6.19）%。NCl-H292細胞凋亡率Hoechst33342+/PI-：20.47 ± 1.46%。處理后48 h，355 nm激光器檢測NB4細胞凋亡率Hoechst33342+/PI-：（33.60 ± 3.75）%。NCl-H292細胞凋亡率Hoechst33342+/PI-：（26.77 ± 1.16）%。407 nm激光器檢測NB4細胞凋亡率Hoechst33342+/PI-：（29.47 ± 2.33）%。NCl-H292細胞凋亡率Hoechst33342+/PI-：（31.47 ± 3.05）%。兩種細胞處理后比處理前凋亡率明顯升高，但355 nm激光器與407 nm激光器檢測的凋亡結果差異不明顯（P＞0.05）。結論　407 nm激光器激發Hoechst33342可檢測細胞凋亡。

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.	徐曉雪, 鄧君, 鄒林樾, 等. 407nm激光流式細胞儀檢測小鼠骨髓側群細胞效果研究[J]. 繼續醫學教育, 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.	王文靜. 側群細胞的研究進展[J]. 國際口腔醫學雜志, 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.

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. 　徐曉雪, 鄧君, 鄒林樾, 等. 407nm激光流式細胞儀檢測小鼠骨髓側群細胞效果研究[J]. 繼續醫學教育, 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. 　王文靜. 側群細胞的研究進展[J]. 國際口腔醫學雜志, 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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