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Correlative Microscopy of Densely Labeled Projection Neurons Using Neural Tracers
Three-dimensional morphological information about neural microcircuits is of high interest in neuroscience, but acquiring this information remains challenging. A promising new correlative technique for brain imaging is array tomography (Micheva and Smith, 2007), in which series of ultrathin brain se...
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Formato: | Texto |
Lenguaje: | English |
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Frontiers Research Foundation
2010
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2912169/ https://www.ncbi.nlm.nih.gov/pubmed/20676237 http://dx.doi.org/10.3389/fnana.2010.00024 |
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author | Oberti, Daniele Kirschmann, Moritz A. Hahnloser, Richard H. R. |
author_facet | Oberti, Daniele Kirschmann, Moritz A. Hahnloser, Richard H. R. |
author_sort | Oberti, Daniele |
collection | PubMed |
description | Three-dimensional morphological information about neural microcircuits is of high interest in neuroscience, but acquiring this information remains challenging. A promising new correlative technique for brain imaging is array tomography (Micheva and Smith, 2007), in which series of ultrathin brain sections are treated with fluorescent antibodies against neurotransmitters and synaptic proteins. Treated sections are repeatedly imaged in the fluorescence light microscope (FLM) and then in the electron microscope (EM). We explore a similar correlative imaging technique in which we differentially label distinct populations of projection neurons, the key routers of electrical signals in the brain. In songbirds, projection neurons can easily be labeled using neural tracers, because the vocal control areas are segregated into separate nuclei. We inject tracers into areas afferent and efferent to the main premotor area for vocal production, HVC, to retrogradely and anterogradely label different classes of projection neurons. We optimize tissue preparation protocols to achieve high fluorescence contrast in the FLM and good ultrastructure in the EM (using osmium tetroxide). Although tracer fluorescence is lost during EM preparation, we localize the tracer molecules after fixation and embedding by using fluorescent antibodies against them. We detect signals mainly in somata and dendrites, allowing us to classify synapses within a single ultrathin section as belonging to a particular type of projection neuron. The use of our method will be to provide statistical information about connectivity among different neuron classes, and to elucidate how signals in the brain are processed and routed among different areas. |
format | Text |
id | pubmed-2912169 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Frontiers Research Foundation |
record_format | MEDLINE/PubMed |
spelling | pubmed-29121692010-07-30 Correlative Microscopy of Densely Labeled Projection Neurons Using Neural Tracers Oberti, Daniele Kirschmann, Moritz A. Hahnloser, Richard H. R. Front Neuroanat Neuroscience Three-dimensional morphological information about neural microcircuits is of high interest in neuroscience, but acquiring this information remains challenging. A promising new correlative technique for brain imaging is array tomography (Micheva and Smith, 2007), in which series of ultrathin brain sections are treated with fluorescent antibodies against neurotransmitters and synaptic proteins. Treated sections are repeatedly imaged in the fluorescence light microscope (FLM) and then in the electron microscope (EM). We explore a similar correlative imaging technique in which we differentially label distinct populations of projection neurons, the key routers of electrical signals in the brain. In songbirds, projection neurons can easily be labeled using neural tracers, because the vocal control areas are segregated into separate nuclei. We inject tracers into areas afferent and efferent to the main premotor area for vocal production, HVC, to retrogradely and anterogradely label different classes of projection neurons. We optimize tissue preparation protocols to achieve high fluorescence contrast in the FLM and good ultrastructure in the EM (using osmium tetroxide). Although tracer fluorescence is lost during EM preparation, we localize the tracer molecules after fixation and embedding by using fluorescent antibodies against them. We detect signals mainly in somata and dendrites, allowing us to classify synapses within a single ultrathin section as belonging to a particular type of projection neuron. The use of our method will be to provide statistical information about connectivity among different neuron classes, and to elucidate how signals in the brain are processed and routed among different areas. Frontiers Research Foundation 2010-06-14 /pmc/articles/PMC2912169/ /pubmed/20676237 http://dx.doi.org/10.3389/fnana.2010.00024 Text en Copyright © 2010 Oberti, Kirschmann and Hahnloser. http://www.frontiersin.org/licenseagreement This is an open-access article subject to an exclusive license agreement between the authors and the Frontiers Research Foundation, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are credited. |
spellingShingle | Neuroscience Oberti, Daniele Kirschmann, Moritz A. Hahnloser, Richard H. R. Correlative Microscopy of Densely Labeled Projection Neurons Using Neural Tracers |
title | Correlative Microscopy of Densely Labeled Projection Neurons Using Neural Tracers |
title_full | Correlative Microscopy of Densely Labeled Projection Neurons Using Neural Tracers |
title_fullStr | Correlative Microscopy of Densely Labeled Projection Neurons Using Neural Tracers |
title_full_unstemmed | Correlative Microscopy of Densely Labeled Projection Neurons Using Neural Tracers |
title_short | Correlative Microscopy of Densely Labeled Projection Neurons Using Neural Tracers |
title_sort | correlative microscopy of densely labeled projection neurons using neural tracers |
topic | Neuroscience |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2912169/ https://www.ncbi.nlm.nih.gov/pubmed/20676237 http://dx.doi.org/10.3389/fnana.2010.00024 |
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