Ciliates are eukaryotic unicellular organisms with complex morphology and developmental processes,including asexual and sexual processes.Conjugation is a form of sexual process that renews genetic materials.However,vi...Ciliates are eukaryotic unicellular organisms with complex morphology and developmental processes,including asexual and sexual processes.Conjugation is a form of sexual process that renews genetic materials.However,visualizing conjugation in ciliates is a challenge due to the complexity and dynamics of the process,while traditional staining methods are often insufficient for the research.This study introduces a new method for visualizing developmental progression in the nuclei during conjugation using Hoechst33342 staining.It describes how to proceed from cell culture,conjugation induction and synchronization,staining preparation,and observation to statistical analysis.The combination of fluorescent staining with the‘volume-fixing'technique eliminates the fixation and dehydration steps,thus reducing the overall operation time to just 20 minutes.This method offers several advantages over traditional staining techniques for studying the nuclei during conjugation.It improves image quality and workflow efficiency and enables real-time observation of live cell states.Potential solutions to challenges that may arise during experimental procedures are introduced and references and guidelines for cytological research are provided in this paper.展开更多
Boehringer Ingelheim(西德的Ingelheim Am Rhein)请求它的竞争对手—Behringwerke(西德的Marburg)在德国血栓溶解剂市场帮助销售血块溶解药—组织纤维蛋白原激活因子(TPA)。Boehringer具有Genentech公司(加利福尼亚州的南旧金山)研制的...Boehringer Ingelheim(西德的Ingelheim Am Rhein)请求它的竞争对手—Behringwerke(西德的Marburg)在德国血栓溶解剂市场帮助销售血块溶解药—组织纤维蛋白原激活因子(TPA)。Boehringer具有Genentech公司(加利福尼亚州的南旧金山)研制的TPA的独家许可权。而Behringwerke(Hoechst AG的一个分公司)则致力于销售链激酶。Boehringer对于销售链激酶无任何特权。终于他们做成了交易,Behringwerke将与Boehringer共同在西德销售TPA。展开更多
文摘目的：利用Hoechst33342/PI双染法和原位末端标记法（TUNEL染色检测纳米二氧化硅（nm-SiO2）对神经细胞凋亡的影响，比较两种检测方法的优缺点。方法：以体外培养的人神经母细胞瘤SK-N-SH为研究对象，15和30 nm粒径的nm-SiO2（剂量为2.5、5、10μg/mL）分别处理细胞24 h，另设1-5 m SiO2组和溶剂对照组，采用Hoechst33342/PI双染法和TUNEL染色检测各处理组对SK-N-SH细胞凋亡的影响。结果：与对照组相比，两种检测方法分析均显示了nm-SiO2处理组SK-N-SH细胞凋亡率显著增加（P〈0.05），且具有尺寸、剂量依赖性，而微米级SiO2对凋亡的影响不显著（P〉0.05）。结论：nm-SiO2能诱导SK-N-SH细胞凋亡。Hoechst33342/PI双染法特异性高，简单易行；TUNEL法灵敏度高，能检测少量的细胞凋亡，但成本较高，两种方法可结合使用以便更加准确的检测神经细胞的凋亡。
基金supported by the National Natural Science Foundation of China(No.32270558)。
文摘Ciliates are eukaryotic unicellular organisms with complex morphology and developmental processes,including asexual and sexual processes.Conjugation is a form of sexual process that renews genetic materials.However,visualizing conjugation in ciliates is a challenge due to the complexity and dynamics of the process,while traditional staining methods are often insufficient for the research.This study introduces a new method for visualizing developmental progression in the nuclei during conjugation using Hoechst33342 staining.It describes how to proceed from cell culture,conjugation induction and synchronization,staining preparation,and observation to statistical analysis.The combination of fluorescent staining with the‘volume-fixing'technique eliminates the fixation and dehydration steps,thus reducing the overall operation time to just 20 minutes.This method offers several advantages over traditional staining techniques for studying the nuclei during conjugation.It improves image quality and workflow efficiency and enables real-time observation of live cell states.Potential solutions to challenges that may arise during experimental procedures are introduced and references and guidelines for cytological research are provided in this paper.
文摘Boehringer Ingelheim(西德的Ingelheim Am Rhein)请求它的竞争对手—Behringwerke(西德的Marburg)在德国血栓溶解剂市场帮助销售血块溶解药—组织纤维蛋白原激活因子(TPA)。Boehringer具有Genentech公司(加利福尼亚州的南旧金山)研制的TPA的独家许可权。而Behringwerke(Hoechst AG的一个分公司)则致力于销售链激酶。Boehringer对于销售链激酶无任何特权。终于他们做成了交易,Behringwerke将与Boehringer共同在西德销售TPA。
