title: Evaluation of neuronal cell death by laser scanning cytometry.
abstract: 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.
authors.affiliation: Unitat de Farmacologia i Farmacognòsia, Facultat de Farmàcia, Universitat de Barcelona, Nucli Universitari de Pedralbes, E-08028, Barcelona, Spain.
authors.forename: Ester
authors.initials: E
authors.lastname: Verdaguer
authors.forename: David
authors.initials: D
authors.lastname: Pubill
authors.forename: Victor
authors.initials: V
authors.lastname: Rimbau
authors.forename: Andrés
authors.initials: A
authors.lastname: Jiménez
authors.forename: Elena
authors.initials: E
authors.lastname: Escubedo
authors.forename: Jordi
authors.initials: J
authors.lastname: Camarasa
authors.forename: Merce
authors.initials: M
authors.lastname: Pallàs
authors.forename: Antoni
authors.initials: A
authors.lastname: Camins
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D016923:Cell Death / Q000187:drug effects* / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D016895:Culture Media, Serum-Free / Q000633:toxicity; D003592:Cytophotometry / Q000295:instrumentation / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D018698:Glutamic Acid / Q000633:toxicity; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation / Q000379:methods*; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D009480:Neuropharmacology / Q000295:instrumentation / Q000379:methods; D009498:Neurotoxins / Q000494:pharmacology*; D009574:Nitro Compounds; D011191:Potassium Deficiency / Q000378:metabolism; D011422:Propionates / Q000633:toxicity; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D016895:Culture Media, Serum-Free; D009498:Neurotoxins; D009574:Nitro Compounds; D011422:Propionates; D018698:Glutamic Acid; C015392:3-nitropropionic acid