When analyzing synaptic connectivity in a brain tissue slice, it is difficult to discern between synapses made by local neurons and those arising from long-range axonal projections. We analyzed a data set of excitatory neurons and inhibitory basket cells reconstructed from cat primary visual cortex in an attempt to provide a quantitative answer to the question: What fraction of cortical synapses is local, and what fraction is mediated by long-range projections? We found an unexpectedly high proportion of nonlocal synapses. For example, 92% of excitatory synapses near the axis of a 200-m-diameter iso-orientation column come from neurons located outside the column, and this fraction remains high-76%-even for an 800-m ocular dominance column. The long-range nature of connectivity has dramatic implications for experiments in cortical tissue slices. Our estimate indicates that in a 300-m-thick section cut perpendicularly to the cortical surface, the number of viable excitatory synapses is reduced to about 10%, and the number of synapses made by inhibitory basket cell axons is reduced to 38%. This uneven reduction in the numbers of excitatory and inhibitory synapses changes the excitationinhibition balance by a factor of 3.8 toward inhibition, and may result in cortical tissue that is less excitable than in vivo. We found that electrophysiological studies conducted in tissue sections may significantly underestimate the extent of cortical connectivity; for example, for some projections, the reported probabilities of finding connected nearby neuron pairs in slices could understate the in vivo probabilities by a factor of 3.axon ͉ connectivity ͉ local ͉ slice W hen examining drawings (1) and 3-dimensional reconstructions (2, 3) of cortical excitatory neurons, it is difficult to evade the impression that most of the excitatory neurons' axons, and, consequently, most of their synapses, are confined to a few hundred micrometer columnar domains surrounding the neurons' somata (Fig. 1A). Only a few branches occasionally escape through the boundaries of the local domain. This interpretation of neuron images can be misleading, however. The few axonal branches that extend beyond the local domain could ramify over large territories (e.g., the entire cortex) and thus could carry a significant fraction of all synapses. Such long-range projections may include interareal and intraareal connections, feedback from higher cortical areas, interhemispheric projections, and feed-forward inputs from subcortical structures. These projections can be easily observed with single neuron or bulk injections of tracers (4-6), but quantifying their fraction is difficult. This is because an accurate estimate of the amount of long-range axons must be made on the scale of the entire cortex, which presents a significant challenge. In practice, long-range axons are truncated in the reconstruction process, and thus their length and richness (density), and the number of synapses that they mediate, cannot be estimated reliably.We devised an approach to deli...