Quantitative investigation into methods for evaluating neocortical slice viability

Abstract

BackgroundIn cortical and hippocampal brain slice experiments, the viability of processed tissue is usually judged by the amplitude of extracellularly-recorded seizure-like event (SLE) activity. Surprisingly, the suitability of this approach for evaluating slice quality has not been objectively studied. Furthermore, a method for gauging the viability of quiescent tissue, in which SLE activity is intentionally suppressed, has not been documented. In this study we undertook to address both of these matters using the zero-magnesium SLE model in neocortical slices.MethodsUsing zero-magnesium SLE activity as the output parameter, we investigated: 1) changes in the pattern (amplitude, frequency and length) of SLE activity as slice health either deteriorated; or was compromised by altering the preparation methodology and; 2) in quiescent tissue, whether the triggering of high frequency field activity following electrode insertion predicted subsequent development of SLE activity — and hence slice viability.ResultsSLE amplitude was the single most important variable correlating with slice viability, with a value less than 50 μV indicative of tissue unlikely to be able to sustain population activity for more than 30–60 minutes. In quiescent slices, an increase in high frequency field activity immediately after electrode insertion predicted the development of SLE activity in 100% of cases. Furthermore, the magnitude of the increase in spectral power correlated with the amplitude of succeeding SLE activity (R2 40.9%, p < 0.0001).ConclusionIn conclusion, the findings confirm that the amplitude of population activity is a suitable field potential parameter for judging brain slice viability — and can be applied independent of the mechanism of tissue activation.