3D multicolor super-resolution imaging offers improved accuracy in neuron tracing.
The connectivity among neurons holds the key to understanding brain function. Mapping neural connectivity in brain circuits requires imaging techniques with high spatial resolution to facilitate neuron tracing and high molecular specificity to mark different cellular and molecular populations. Here,...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2012-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3265519?pdf=render |
| _version_ | 1818273924341825536 |
|---|---|
| author | Melike Lakadamyali Hazen Babcock Mark Bates Xiaowei Zhuang Jeff Lichtman |
| author_facet | Melike Lakadamyali Hazen Babcock Mark Bates Xiaowei Zhuang Jeff Lichtman |
| author_sort | Melike Lakadamyali |
| collection | DOAJ |
| description | The connectivity among neurons holds the key to understanding brain function. Mapping neural connectivity in brain circuits requires imaging techniques with high spatial resolution to facilitate neuron tracing and high molecular specificity to mark different cellular and molecular populations. Here, we tested a three-dimensional (3D), multicolor super-resolution imaging method, stochastic optical reconstruction microscopy (STORM), for tracing neural connectivity using cultured hippocampal neurons obtained from wild-type neonatal rat embryos as a model system. Using a membrane specific labeling approach that improves labeling density compared to cytoplasmic labeling, we imaged neural processes at 44 nm 2D and 116 nm 3D resolution as determined by considering both the localization precision of the fluorescent probes and the Nyquist criterion based on label density. Comparison with confocal images showed that, with the currently achieved resolution, we could distinguish and trace substantially more neuronal processes in the super-resolution images. The accuracy of tracing was further improved by using multicolor super-resolution imaging. The resolution obtained here was largely limited by the label density and not by the localization precision of the fluorescent probes. Therefore, higher image resolution, and thus higher tracing accuracy, can in principle be achieved by further improving the label density. |
| first_indexed | 2024-12-12T22:05:41Z |
| format | Article |
| id | doaj.art-3c1fd2f1e4f64da4b37df9ab24dc4995 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-12T22:05:41Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-3c1fd2f1e4f64da4b37df9ab24dc49952022-12-22T00:10:23ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0171e3082610.1371/journal.pone.00308263D multicolor super-resolution imaging offers improved accuracy in neuron tracing.Melike LakadamyaliHazen BabcockMark BatesXiaowei ZhuangJeff LichtmanThe connectivity among neurons holds the key to understanding brain function. Mapping neural connectivity in brain circuits requires imaging techniques with high spatial resolution to facilitate neuron tracing and high molecular specificity to mark different cellular and molecular populations. Here, we tested a three-dimensional (3D), multicolor super-resolution imaging method, stochastic optical reconstruction microscopy (STORM), for tracing neural connectivity using cultured hippocampal neurons obtained from wild-type neonatal rat embryos as a model system. Using a membrane specific labeling approach that improves labeling density compared to cytoplasmic labeling, we imaged neural processes at 44 nm 2D and 116 nm 3D resolution as determined by considering both the localization precision of the fluorescent probes and the Nyquist criterion based on label density. Comparison with confocal images showed that, with the currently achieved resolution, we could distinguish and trace substantially more neuronal processes in the super-resolution images. The accuracy of tracing was further improved by using multicolor super-resolution imaging. The resolution obtained here was largely limited by the label density and not by the localization precision of the fluorescent probes. Therefore, higher image resolution, and thus higher tracing accuracy, can in principle be achieved by further improving the label density.http://europepmc.org/articles/PMC3265519?pdf=render |
| spellingShingle | Melike Lakadamyali Hazen Babcock Mark Bates Xiaowei Zhuang Jeff Lichtman 3D multicolor super-resolution imaging offers improved accuracy in neuron tracing. PLoS ONE |
| title | 3D multicolor super-resolution imaging offers improved accuracy in neuron tracing. |
| title_full | 3D multicolor super-resolution imaging offers improved accuracy in neuron tracing. |
| title_fullStr | 3D multicolor super-resolution imaging offers improved accuracy in neuron tracing. |
| title_full_unstemmed | 3D multicolor super-resolution imaging offers improved accuracy in neuron tracing. |
| title_short | 3D multicolor super-resolution imaging offers improved accuracy in neuron tracing. |
| title_sort | 3d multicolor super resolution imaging offers improved accuracy in neuron tracing |
| url | http://europepmc.org/articles/PMC3265519?pdf=render |
| work_keys_str_mv | AT melikelakadamyali 3dmulticolorsuperresolutionimagingoffersimprovedaccuracyinneurontracing AT hazenbabcock 3dmulticolorsuperresolutionimagingoffersimprovedaccuracyinneurontracing AT markbates 3dmulticolorsuperresolutionimagingoffersimprovedaccuracyinneurontracing AT xiaoweizhuang 3dmulticolorsuperresolutionimagingoffersimprovedaccuracyinneurontracing AT jefflichtman 3dmulticolorsuperresolutionimagingoffersimprovedaccuracyinneurontracing |