Evaluation of neuronal cell death by laser scanning cytometry.
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.
Verdaguer
Ester
E
Unitat de Farmacologia i Farmacognòsia, Facultat de Farmàcia, Universitat de Barcelona, Nucli Universitari de Pedralbes, E-08028, Barcelona, Spain.
Pubill
David
D
Rimbau
Victor
V
Jiménez
Andrés
A
Escubedo
Elena
E
Camarasa
Jordi
J
Pallàs
Merce
M
Camins
Antoni
A
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
D016895:Culture Media, Serum-Free; D009498:Neurotoxins; D009574:Nitro Compounds; D011422:Propionates; D018698:Glutamic Acid; C015392:3-nitropropionic acid