Page 111 - Live-cellanalysis handbook

Neuronal Activity Assay

Long-term quantification of synaptic
activity and network connectivity

Introduction

The most fundamental function of the nervous system is the techniques, such as patch clamp measurements and multi-
transmission and integration of information via electrical and electrode arrays (MEA), may provide exquisite resolution of
chemical signals that pass from axon termini to receiving dendrites electrical changes, however these measurements are often
of neighboring neurons. In order to coordinate complex processes, derived at a single point in time without confirmation of cell
this primary function of integration and propagation of cellular morphology. Furthermore, MEA methods require neuronal cultures
signals must be achieved. Activity measurements are therefore to be grown at high densities which sacrifices representation
critical for characterizing the generation and maturation of of in vivo conditions. Analysis of calcium oscillations offers the
neuronal networks to gain functional insights into relevant opportunity to analyze morphological changes associated with
neuronal models and disease states. Model systems gaining the neuronal activity, however also fail to capture long-term changes
most traction in the field of neuroscience are human induced of neuronal activity and cannot detect the formation of a mature,
pluripotent stem cell (hiPSC)-derived neurons, which offer the connected neuronal network.
opportunity to study basic neuronal development as well as
representative disease conditions as they differentiate, mature Live-cell analysis alleviates many of these challenges by allowing
and become functionally active. However, current technologies for chronic evaluation (over days, weeks, or months) of neuronal
have limited ability to generate the information needed to develop activity and connectivity from thousands of cells per well, via
these models and determine when they become functionally non-destructive, repeated imaging of the same sample in a
active. physiologically relevant environment. In this chapter, we illustrate
how the neuronal activity assay using the IncuCyte Live-Cell
®
Traditionally, neuronal activity measurements are made using Analysis System in conjunction with fit-for-purpose software tools
sophisticated microelectrode electrophysiology techniques or and a non-perturbing reagent enables kinetic quantification of
microscopic analysis of calcium oscillations. Electrophysiology activity and connectivity, at microplate scale.

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