Reconstruction of genetically identified neurons imaged by serial-section electron microscopy
Resolving patterns of synaptic connectivity in neural circuits currently requires serial section electron microscopy. However, complete circuit reconstruction is prohibitively slow and may not be necessary for many purposes such as comparing neuronal structure and connectivity among multiple animals...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2016-07-01
|
| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/15015 |
| _version_ | 1811181014723067904 |
|---|---|
| author | Maximilian Joesch David Mankus Masahito Yamagata Ali Shahbazi Richard Schalek Adi Suissa-Peleg Markus Meister Jeff W Lichtman Walter J Scheirer Joshua R Sanes |
| author_facet | Maximilian Joesch David Mankus Masahito Yamagata Ali Shahbazi Richard Schalek Adi Suissa-Peleg Markus Meister Jeff W Lichtman Walter J Scheirer Joshua R Sanes |
| author_sort | Maximilian Joesch |
| collection | DOAJ |
| description | Resolving patterns of synaptic connectivity in neural circuits currently requires serial section electron microscopy. However, complete circuit reconstruction is prohibitively slow and may not be necessary for many purposes such as comparing neuronal structure and connectivity among multiple animals. Here, we present an alternative strategy, targeted reconstruction of specific neuronal types. We used viral vectors to deliver peroxidase derivatives, which catalyze production of an electron-dense tracer, to genetically identify neurons, and developed a protocol that enhances the electron-density of the labeled cells while retaining the quality of the ultrastructure. The high contrast of the marked neurons enabled two innovations that speed data acquisition: targeted high-resolution reimaging of regions selected from rapidly-acquired lower resolution reconstruction, and an unsupervised segmentation algorithm. This pipeline reduces imaging and reconstruction times by two orders of magnitude, facilitating directed inquiry of circuit motifs. |
| first_indexed | 2024-04-11T09:11:13Z |
| format | Article |
| id | doaj.art-1a7b7e801de74d13a4cf4f2b304eca2b |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T09:11:13Z |
| publishDate | 2016-07-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-1a7b7e801de74d13a4cf4f2b304eca2b2022-12-22T04:32:29ZengeLife Sciences Publications LtdeLife2050-084X2016-07-01510.7554/eLife.15015Reconstruction of genetically identified neurons imaged by serial-section electron microscopyMaximilian Joesch0David Mankus1Masahito Yamagata2https://orcid.org/0000-0001-8193-2931Ali Shahbazi3Richard Schalek4Adi Suissa-Peleg5Markus Meister6https://orcid.org/0000-0003-2136-6506Jeff W Lichtman7Walter J Scheirer8Joshua R Sanes9https://orcid.org/0000-0001-8926-8836Center for Brain Science, Harvard University, Cambridge, United States; Department of Molecular and Cellular Biology, Harvard University, Cambridge, United StatesCenter for Brain Science, Harvard University, Cambridge, United States; Department of Molecular and Cellular Biology, Harvard University, Cambridge, United StatesCenter for Brain Science, Harvard University, Cambridge, United States; Department of Molecular and Cellular Biology, Harvard University, Cambridge, United StatesUniversity of Notre Dame, Notre Dame, United StatesCenter for Brain Science, Harvard University, Cambridge, United States; Department of Molecular and Cellular Biology, Harvard University, Cambridge, United StatesSchool of Engineering and Applied Sciences, Harvard University, Cambridge, United StatesDivision of Biology, California Institute of Technology, Pasadena, United StatesCenter for Brain Science, Harvard University, Cambridge, United States; Department of Molecular and Cellular Biology, Harvard University, Cambridge, United StatesUniversity of Notre Dame, Notre Dame, United StatesCenter for Brain Science, Harvard University, Cambridge, United States; Department of Molecular and Cellular Biology, Harvard University, Cambridge, United StatesResolving patterns of synaptic connectivity in neural circuits currently requires serial section electron microscopy. However, complete circuit reconstruction is prohibitively slow and may not be necessary for many purposes such as comparing neuronal structure and connectivity among multiple animals. Here, we present an alternative strategy, targeted reconstruction of specific neuronal types. We used viral vectors to deliver peroxidase derivatives, which catalyze production of an electron-dense tracer, to genetically identify neurons, and developed a protocol that enhances the electron-density of the labeled cells while retaining the quality of the ultrastructure. The high contrast of the marked neurons enabled two innovations that speed data acquisition: targeted high-resolution reimaging of regions selected from rapidly-acquired lower resolution reconstruction, and an unsupervised segmentation algorithm. This pipeline reduces imaging and reconstruction times by two orders of magnitude, facilitating directed inquiry of circuit motifs.https://elifesciences.org/articles/15015connectomicsperoxidaseelectron microscopyreconstruction |
| spellingShingle | Maximilian Joesch David Mankus Masahito Yamagata Ali Shahbazi Richard Schalek Adi Suissa-Peleg Markus Meister Jeff W Lichtman Walter J Scheirer Joshua R Sanes Reconstruction of genetically identified neurons imaged by serial-section electron microscopy eLife connectomics peroxidase electron microscopy reconstruction |
| title | Reconstruction of genetically identified neurons imaged by serial-section electron microscopy |
| title_full | Reconstruction of genetically identified neurons imaged by serial-section electron microscopy |
| title_fullStr | Reconstruction of genetically identified neurons imaged by serial-section electron microscopy |
| title_full_unstemmed | Reconstruction of genetically identified neurons imaged by serial-section electron microscopy |
| title_short | Reconstruction of genetically identified neurons imaged by serial-section electron microscopy |
| title_sort | reconstruction of genetically identified neurons imaged by serial section electron microscopy |
| topic | connectomics peroxidase electron microscopy reconstruction |
| url | https://elifesciences.org/articles/15015 |
| work_keys_str_mv | AT maximilianjoesch reconstructionofgeneticallyidentifiedneuronsimagedbyserialsectionelectronmicroscopy AT davidmankus reconstructionofgeneticallyidentifiedneuronsimagedbyserialsectionelectronmicroscopy AT masahitoyamagata reconstructionofgeneticallyidentifiedneuronsimagedbyserialsectionelectronmicroscopy AT alishahbazi reconstructionofgeneticallyidentifiedneuronsimagedbyserialsectionelectronmicroscopy AT richardschalek reconstructionofgeneticallyidentifiedneuronsimagedbyserialsectionelectronmicroscopy AT adisuissapeleg reconstructionofgeneticallyidentifiedneuronsimagedbyserialsectionelectronmicroscopy AT markusmeister reconstructionofgeneticallyidentifiedneuronsimagedbyserialsectionelectronmicroscopy AT jeffwlichtman reconstructionofgeneticallyidentifiedneuronsimagedbyserialsectionelectronmicroscopy AT walterjscheirer reconstructionofgeneticallyidentifiedneuronsimagedbyserialsectionelectronmicroscopy AT joshuarsanes reconstructionofgeneticallyidentifiedneuronsimagedbyserialsectionelectronmicroscopy |