Verdaguer Ester, Pubill David, Rimbau Victor, Jiménez Andrés, Escubedo Elena, Camarasa Jordi, Pallàs Merce, Camins Antoni
Unitat de Farmacologia i Farmacognòsia, Facultat de Farmàcia, Universitat de Barcelona, Nucli Universitari de Pedralbes, E-08028, Barcelona, Spain.
Brain Res Brain Res Protoc. 2002 Feb;9(1):41-8. doi: 10.1016/s1385-299x(01)00135-0.
We developed a method in which laser scanning cytometry (LSC) is applied to evaluate cell viability. Neuronal cell death induced by glutamic acid, serum potassium deprivation and 3-nitropropionic acid was studied in cerebellar granule cells by neutral red assay (NR) and LSC, using propidium iodide (PI) as fluorescent dye. PI labeled the nuclei of dead neurons and increased fluorescence was measured using a laser scanning cytometer. Similar levels of damage for each injury were detected by NR or LSC. The protocol presented here, provides a fast and sensitive assay for the analysis of neuronal viability using LSC, and can be used to study new neuroprotective drugs in neuronal cell cultures.
我们开发了一种应用激光扫描细胞术（LSC）评估细胞活力的方法。通过中性红试验（NR）和LSC，使用碘化丙啶（PI）作为荧光染料，研究了谷氨酸、血清钾缺乏和3-硝基丙酸诱导的小脑颗粒细胞神经元细胞死亡。PI标记死亡神经元的细胞核，并使用激光扫描细胞仪测量增加的荧光。通过NR或LSC检测到每种损伤的相似损伤水平。本文介绍的方案提供了一种使用LSC分析神经元活力的快速灵敏检测方法，可用于研究神经元细胞培养中的新型神经保护药物。
