Monitoring Population Membrane Potential Signals from Neocortex

Voltage-sensitive dyes (VSDs) and optical imaging are useful tools for studying spatiotemporal patterns of population neuronal activity in cortex. Because fast VSDs respond to membrane potential changes with microsecond temporal resolution, these are better suited than calcium indicators for recording rapid neural signals. Here we describe methods for using a 464 element photodiode array and fast VSDs to record signals ranging from large scale network activity in brain slices and in vivo mammalian preparations with sensitivity comparable to local field potential (LFP) recordings. With careful control of dye bleaching and phototoxicity, long recording times can be achieved. Absorption dyes have less photo-toxicity than fluorescent dyes. In brain slices, the total recording time in each slice can be 1,000-2,000 s, which can be divided into hundreds of short recording trials over several hours. In intact brains when fluorescent dyes are used, reduced light intensity can also increase recording time. In this chapter, we will discuss technical details for the methods to achieve reliable VSD imaging with high sensitivity and long recording time.