Quantitative Live/Dead Viability Assay

Cell death was determined by Live/Dead viability assay, as described previously (Doria et al., 2013 (link)). Briefly, neurons were stained with 2 μM calcein acetoxymethyl ester (AM) and 2 μM ethidium homodimer‐1 for 15 min and the fractions of live (calcein AM positive) and dead (ethidium homodimer-1 positive) cells were determined by microscopy. Neurons were visualized by fluorescence microscopy FLoid® Cell Imaging Station (Thermo Scientific, Waltham, MA, USA) and scored by a blinded observer. A minimum of 300 cells were analyzed per well in triplicates using the ImageJ™ software. Dead cells were expressed as a percentage of the total number of cells.

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Olmo I.G., Olmo R.P., Gonçalves A.N., Pires R.G., Marques J.T, & Ribeiro F.M. (2021). High-Throughput Sequencing of BACHD Mice Reveals Upregulation of Neuroprotective miRNAs at the Pre-Symptomatic Stage of Huntington’s Disease. ASN NEURO, 13, 17590914211009857.

Publication 2021

FLoid® Cell Imaging Station

A 300 Assay Calcein acetoxymethyl ester Cell death Cells Ethidium homodimer 1 Fluorescence microscopy Microscopy Neurons

Corresponding Organization : Universidade Federal de Minas Gerais

Other organizations : Université de Strasbourg, Institut de Biologie Moléculaire et Cellulaire, Inserm, Centre National de la Recherche Scientifique, Universidade de São Paulo, Universidade Federal do Espírito Santo

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Variable analysis

independent variables

Neuronal treatment conditions (unspecified)

dependent variables

Percentage of dead cells

control variables

Microscopy visualization method (FLoid® Cell Imaging Station)
Cell counting method (ImageJ™ software)
Staining concentrations (2 μM calcein AM and 2 μM ethidium homodimer-1)
Staining duration (15 min)
Observer blinding

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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