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Q:Hoechst33342/PI双染试剂盒原理及结果判断?
A:Hoechst33342/PI主要是区分凋亡细胞和坏死细胞的,细胞发生凋亡时,染色质会固缩。Hoechst33342可以穿透细胞膜,染色后凋亡细胞荧光会比正常细胞明显增强。碘化丙啶(PI)不能穿透细胞膜,对于具有完整细胞膜的正常细胞或凋亡细胞不能染色。而对于坏死细胞,其细胞膜的完整性丧失,碘化丙啶(PI)可以染色坏死细胞。
正常细胞:弱红色荧光+弱蓝色荧光
凋亡细胞:弱红色荧光+强蓝色荧光
坏死细胞:强红色荧光+弱蓝色荧光
Q: Hoechst 33342/PI 双染试剂盒中两种染料浓度是多少?
A: Hoechst:1mg/mL, PI浓度:200ug/mL。
Q: Hoechst 33342在4度还是37度染色合适?
A: Hoechst 33342在37度染色更合适,4度染色可能导致细胞核不能被完全染色。
Published citations
[1] Zi Q, Ding W, Sun C, et al. On-chip label-free determination of cell survival rate. Biosens Bioelectron. 2020;148:111820. doi:10.1016/j.bios.2019.111820(IF:9.518)
[2] Li X, Fu G, Zhang L, et al. Assay establishment and validation of a high-throughput organoid-based drug screening platform. Stem Cell Res Ther. 2022;13(1):219. Published 2022 May 26. doi:10.1186/s13287-022-02902-3(IF:6.832)
[3] Gao X, Liu S, Tan L, et al. Estrogen Receptor α Regulates Metabolic-Associated Fatty Liver Disease by Targeting NLRP3-GSDMD Axis-Mediated Hepatocyte Pyroptosis. J Agric Food Chem. 2021;69(48):14544-14556. doi:10.1021/acs.jafc.1c05400(IF:5.279)
[4] Liu S, Xu DS, Ma JL, Huang P, Wu D, Ren LQ. LncRNA H19 Mitigates Oxidized Low-Density Lipoprotein Induced Pyroptosis via Caspase-1 in Raw 264.7 Cells. Inflammation. 2021;44(6):2407-2418. doi:10.1007/s10753-021-01511-1(IF:4.092)
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