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A report on digitised neuronal tracing method to study neurons in their entirety

Conventional camera lucida (CL) aided neuronal tracing technique for studying neural plasticity is a demanding procedure. Stereo Investigator-Neurolucida enabled neuronal tracing system is not accessible to all researchers. This necessitates alternate simple and less challenging digitised neuronal t...

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Detalles Bibliográficos
Autores principales: Ali, Zeeshan, Sivakumar, G., Prabhu, Krishnamoorthi, Suryavanshi, Chinmay Ajit, Narayanan, Sareesh Naduvil
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9111970/
https://www.ncbi.nlm.nih.gov/pubmed/35592463
http://dx.doi.org/10.1016/j.mex.2022.101715
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author Ali, Zeeshan
Sivakumar, G.
Prabhu, Krishnamoorthi
Suryavanshi, Chinmay Ajit
Narayanan, Sareesh Naduvil
author_facet Ali, Zeeshan
Sivakumar, G.
Prabhu, Krishnamoorthi
Suryavanshi, Chinmay Ajit
Narayanan, Sareesh Naduvil
author_sort Ali, Zeeshan
collection PubMed
description Conventional camera lucida (CL) aided neuronal tracing technique for studying neural plasticity is a demanding procedure. Stereo Investigator-Neurolucida enabled neuronal tracing system is not accessible to all researchers. This necessitates alternate simple and less challenging digitised neuronal tracing methods. This report describes a novel digitised neuronal tracing method using widefield microscopy, and its effectiveness is compared with the traditional camera lucida aided neuronal tracing method. Golgi-Cox stained hippocampal cornu ammonis area-3 (CA3) pyramidal neuron photomicrographs were serially captured at a depth of every 2µm in the z-axis by a wide field microscope from the point of appearance to the disappearance. These images were stacked along the axis perpendicular to the image plane to reconstruct the neuron in its entirety, digitally traced and dendritic quantification was performed using open source software. The same neurons were manually traced using camera lucida, and Sholl analysis was done manually to quantify the dendritic arborisation pattern. The dendritic quantification data were not significantly different in both methods. Hence, the technology-enabled, less demanding, and equally accurate neuronal tracing can be adopted instead of manual tracing and analysis of neurons. • A simple digitised neuronal tracing method is described. • It is fast, rigorous, and comparable to traditional tracing techniques. • Helps the researcher to repeatedly probe data to reduce errors.
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spelling pubmed-91119702022-05-18 A report on digitised neuronal tracing method to study neurons in their entirety Ali, Zeeshan Sivakumar, G. Prabhu, Krishnamoorthi Suryavanshi, Chinmay Ajit Narayanan, Sareesh Naduvil MethodsX Method Article Conventional camera lucida (CL) aided neuronal tracing technique for studying neural plasticity is a demanding procedure. Stereo Investigator-Neurolucida enabled neuronal tracing system is not accessible to all researchers. This necessitates alternate simple and less challenging digitised neuronal tracing methods. This report describes a novel digitised neuronal tracing method using widefield microscopy, and its effectiveness is compared with the traditional camera lucida aided neuronal tracing method. Golgi-Cox stained hippocampal cornu ammonis area-3 (CA3) pyramidal neuron photomicrographs were serially captured at a depth of every 2µm in the z-axis by a wide field microscope from the point of appearance to the disappearance. These images were stacked along the axis perpendicular to the image plane to reconstruct the neuron in its entirety, digitally traced and dendritic quantification was performed using open source software. The same neurons were manually traced using camera lucida, and Sholl analysis was done manually to quantify the dendritic arborisation pattern. The dendritic quantification data were not significantly different in both methods. Hence, the technology-enabled, less demanding, and equally accurate neuronal tracing can be adopted instead of manual tracing and analysis of neurons. • A simple digitised neuronal tracing method is described. • It is fast, rigorous, and comparable to traditional tracing techniques. • Helps the researcher to repeatedly probe data to reduce errors. Elsevier 2022-04-29 /pmc/articles/PMC9111970/ /pubmed/35592463 http://dx.doi.org/10.1016/j.mex.2022.101715 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Method Article
Ali, Zeeshan
Sivakumar, G.
Prabhu, Krishnamoorthi
Suryavanshi, Chinmay Ajit
Narayanan, Sareesh Naduvil
A report on digitised neuronal tracing method to study neurons in their entirety
title A report on digitised neuronal tracing method to study neurons in their entirety
title_full A report on digitised neuronal tracing method to study neurons in their entirety
title_fullStr A report on digitised neuronal tracing method to study neurons in their entirety
title_full_unstemmed A report on digitised neuronal tracing method to study neurons in their entirety
title_short A report on digitised neuronal tracing method to study neurons in their entirety
title_sort report on digitised neuronal tracing method to study neurons in their entirety
topic Method Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9111970/
https://www.ncbi.nlm.nih.gov/pubmed/35592463
http://dx.doi.org/10.1016/j.mex.2022.101715
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