on optical detection of densely labeled synapses in neuropil and mapping connectivity with combinatorially multiplexed fluorescent synaptic markers密集的光学检测标记突触神经纤维网和映射连接组合多路复用荧光突触标记.pdfVIP

On Optical Detection of Densely Labeled Synapses in Neuropil and Mapping Connectivity with Combinatorially Multiplexed Fluorescent Synaptic Markers Yuriy Mishchenko* Department of Statistics and Center for Theoretical Neuroscience, Columbia University, New York, New York, United States of America Abstract We propose a new method for mapping neural connectivity optically, by utilizing Cre/Lox system Brainbow to tag synapses of different neurons with random mixtures of different fluorophores, such as GFP, YFP, etc., and then detecting patterns of fluorophores at different synapses using light microscopy (LM). Such patterns will immediately report the pre- and post- synaptic cells at each synaptic connection, without tracing neural projections from individual synapses to corresponding cell bodies. We simulate fluorescence from a population of densely labeled synapses in a block of hippocampal neuropil, completely reconstructed from electron microscopy data, and show that high-end LM is able to detect such patterns with over 95% accuracy. We conclude, therefore, that with the described approach neural connectivity in macroscopically large neural circuits can be mapped with great accuracy, in scalable manner, using fast optical tools, and straightforward image processing. Relying on an electron microscopy dataset, we also derive and explicitly enumerate the conditions that should be met to allow synaptic connectivity studies with high-resolution optical tools. Citation: Mishchenko Y (2010) On Optical Detection of Densely Labeled Synapses in Neuropil and Mapping Connectivity with Combinatorially Multiplexed Fluorescent Synaptic Markers. PLoS ONE 5(1): e8853. doi:10.1371/journal.pone.0008853 Editor: David Finkelstein, The Mental Health Research Institute of Victoria, Australia Received October 19, 2009; Accepted December 24, 2009; Published January 22,