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Commentary Decades of research have helped to characterize many electrophysiological patterns in normal and in epileptic brain tissue. However, this characterization has primarily been on the scale of either a very large or very small number of neurons. Neurophysiological recordings—from EEG down to local fields—record electrical phenomena occurring in a large population of neurons: spikes, slow waves, seizures, etc. While these phenomena form the basis of clinical decision making, little is known about their complex dynamics at the cellular level. On the other side of the spectrum, techniques such as patch clamping and spike sorting monitor individual action potentials, but typically follow fewer than 10 cells at a time. Multi-electrode arrays can monitor up to 100 cells but become technically challenging beyond that number. One method to bridge this gap and monitor action potentials from larger numbers of cells simultaneously is to use voltageor calciumsensitive dyes. The key to this technology is that the number of recorded cells depends upon optics and the camera resolution rather than electrode placement. Different forms of this technology have been under development for several years, but it has been challenging to apply it to the fast waveforms seen in epilepsy. In this article, Takano and colleagues use a combination of fast confocal microscopy and multicellular calcium imaging to monitor a large number of neurons during network bursts, then use sophisticated statistics to process the data. They are able to track the firing order of action potentials of individual cells during two different types of bursting activity, and find intriguing differences between these bursts. Over the past 2 decades, several technologies have been developed that strive to monitor the activity of neuronal networks in the brain. Most of these exploit advanced imaging technology: fMRI, SPECT, PET, MEG, and microscopic imaging of voltage-sensitive dyes. These technologies have been revolutionary but often have limited spatial and temporal resolution. Relatively brief events such as epileptic spikes are fast and small enough that it has been very difficult to investigate network behavior at the cellular level. A more recent development is functional multicellular (or multineuronal) calcium imaging (fMCI) (1). In this technique, which has been under development for just over a decade (2), a calcium-sensitive fluorescence indicator is bulk-loaded across a large area of brain cortex, and it can be performed in vivo (3). The fluorescent molecule is designed so that intracellular concentration rises to much higher levels than in the extracellular space, which produces good signal fidelity. The intracellular indicator changes fluorescence very quickly and strongly in response to calcium influx, allowing imaging of action potentials in individual cells. The current work uses this technology to analyze population firing during epileptiform bursts in the hippocampal Deterministic and Stochastic Neuronal Contributions to Distinct Synchronous CA3 Network Bursts.

Dyeing to be Fired: Firing Order Distinguishes Two Types of Bursting Activity