期刊
FRONTIERS IN NEUROANATOMY
卷 8, 期 -, 页码 -出版社
FRONTIERS MEDIA SA
DOI: 10.3389/fnana.2014.00065
关键词
morphology; reconstruction; cell-type classification; multicompartmental modeling; interneurons; 2-photon imaging; Neurolucida; neocortex
资金
- BBSRC Industrial CASE studentship [BB/H016600/1]
- Scientifica, EU FP7 Future Emergent Technologies [243914]
- European Union [269921, CIHR OG 126137]
- NSERC [DG 418546-12]
- MRC [G0700188] Funding Source: UKRI
- Medical Research Council [G0700188] Funding Source: researchfish
Accurate 3D reconstruction of neurons is vital for applications linking anatomy and physiology. Reconstructions are typically created using Neurolucida after biocytin histology (BH). An alternative inexpensive and fast method is to use freeware such as Neuromantic to reconstruct from fluorescence imaging (El) stacks acquired using 2-photon laser-scanning microscopy during physiological recording. We compare these two methods with respect to morphometry, cell classification, and multicompartmental modeling in the NEURON simulation environment. Quantitative morphological analysis of the same cells reconstructed using both methods reveals that whilst biocytin reconstructions facilitate tracing of more distal collaterals, both methods are comparable in representing the overall morphology: automated clustering of reconstructions from both methods successfully separates neocortical basket cells from pyramidal cells but not BH from Fl reconstructions. BH reconstructions suffer more from tissue shrinkage and compression artifacts than Fl reconstructions do. Fl reconstructions, on the other hand, consistently have larger process diameters. Consequently, significant differences in NEURON modeling of excitatory post-synaptic potential (EPSP) forward propagation are seen between the two methods, with Fl reconstructions exhibiting smaller depolarizations. Simulated action potential backpropagation (bAP), however, is indistinguishable between reconstructions obtained with the two methods. In our hands, BH reconstructions are necessary for NEURON modeling and detailed morphological tracing, and thus remain state of the art, although they are more labor intensive, more expensive, and suffer from a higher failure rate due to the occasional poor outcome of histological processing. However, for a subset of anatomical applications such as cell type identification, Fl reconstructions are superior, because of indistinguishable classification performance with greater ease of use, essentially 100% success rate, and lower cost.
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