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  Transmitted light brightfield mosaic microscopy for three-dimensional tracing of single neuron morphology

Oberlaender, M., Bruno, R. M., Sakmann, B., & Broser, P. J. (2007). Transmitted light brightfield mosaic microscopy for three-dimensional tracing of single neuron morphology. Journal of Biomedical Optics, 12(6): 064029, pp. 1-19. doi:10.1117/1.2815693.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-ACAE-A Version Permalink: http://hdl.handle.net/21.11116/0000-0006-DB69-E
Genre: Journal Article
Alternative Title : Transmitted light brightfield mosaic microscopy for three-dimensional tracing of single neuron morphologyneuron morphology

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https://dx.doi.org/10.1117/1.2815693 (Any fulltext)
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 Creators:
Oberlaender, Marcel1, Author              
Bruno, Randy M.1, Author              
Sakmann, Bert1, Author              
Broser, Philip Julian1, Author              
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1Department of Cell Physiology, Max Planck Institute for Medical Research, Max Planck Society, ou_1497701              

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Free keywords: three-dimensional automatic reconstruction; three-dimensional mosaic microscopy; three-dimensional neuron morphology; large data processing; axonal morphology; axons
 Abstract: A fundamental challenge in neuroscience is the determination of the three-dimensional (3D) morphology of neurons in the cortex. Here we describe a semiautomated method to trace single biocytin-filled neurons using a transmitted light brightfield microscope. The method includes 3D tracing of dendritic trees and axonal arbors from image stacks of serial 100-microm-thick tangential brain sections. Key functionalities include mosaic scanning and optical sectioning, high-resolution image restoration, and fast, parallel computing for neuron tracing. The mosaic technique compensates for the limited field of view at high magnification, allowing the acquisition of high-resolution image stacks on a scale of millimeters. The image restoration by deconvolution is based on experimentally verified assumptions about the optical system. Restoration yields a significant improvement of signal-to-noise ratio and resolution of neuronal structures in the image stack. Application of local threshold and thinning filters result in a 3D graph representation of dendrites and axons in a section. The reconstructed branches are then manually edited and aligned. Branches from adjacent sections are spliced, resulting in a complete 3D reconstruction of a neuron. A comparison with 3D reconstructions from manually traced neurons shows that the semiautomated system is a fast and reliable alternative to the manual tracing systems currently available.

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Language(s): eng - English
 Dates: 2007-06-192007-04-022007-07-092007-12-282007-12-28
 Publication Status: Published in print
 Pages: 19
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Degree: -

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Title: Journal of Biomedical Optics
Source Genre: Journal
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Publ. Info: Bellingham, WA : Published by SPIE--the International Society for Optical Engineering in cooperation with International Biomedical Optics Society
Pages: - Volume / Issue: 12 (6) Sequence Number: 064029 Start / End Page: 1 - 19 Identifier: ISSN: 1083-3668
CoNE: https://pure.mpg.de/cone/journals/resource/954925607859