Cargando…
Sources of widefield fluorescence from the brain
Widefield fluorescence microscopy is used to monitor the spiking of populations of neurons in the brain. Widefield fluorescence can originate from indicator molecules at all depths in cortex and the relative contributions from somata, dendrites, and axons are often unknown. Here, I simulate widefiel...
Autor principal: | |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7647397/ https://www.ncbi.nlm.nih.gov/pubmed/33155981 http://dx.doi.org/10.7554/eLife.59841 |
_version_ | 1783606902804971520 |
---|---|
author | Waters, Jack |
author_facet | Waters, Jack |
author_sort | Waters, Jack |
collection | PubMed |
description | Widefield fluorescence microscopy is used to monitor the spiking of populations of neurons in the brain. Widefield fluorescence can originate from indicator molecules at all depths in cortex and the relative contributions from somata, dendrites, and axons are often unknown. Here, I simulate widefield illumination and fluorescence collection and determine the main sources of fluorescence for several GCaMP mouse lines. Scattering strongly affects illumination and collection. One consequence is that illumination intensity is greatest ~300–400 µm below the pia, not at the brain surface. Another is that fluorescence from a source deep in cortex may extend across a diameter of 3–4 mm at the brain surface, severely limiting lateral resolution. In many mouse lines, the volume of tissue contributing to fluorescence extends through the full depth of cortex and fluorescence at most surface locations is a weighted average across multiple cortical columns and often more than one cortical area. |
format | Online Article Text |
id | pubmed-7647397 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-76473972020-11-09 Sources of widefield fluorescence from the brain Waters, Jack eLife Neuroscience Widefield fluorescence microscopy is used to monitor the spiking of populations of neurons in the brain. Widefield fluorescence can originate from indicator molecules at all depths in cortex and the relative contributions from somata, dendrites, and axons are often unknown. Here, I simulate widefield illumination and fluorescence collection and determine the main sources of fluorescence for several GCaMP mouse lines. Scattering strongly affects illumination and collection. One consequence is that illumination intensity is greatest ~300–400 µm below the pia, not at the brain surface. Another is that fluorescence from a source deep in cortex may extend across a diameter of 3–4 mm at the brain surface, severely limiting lateral resolution. In many mouse lines, the volume of tissue contributing to fluorescence extends through the full depth of cortex and fluorescence at most surface locations is a weighted average across multiple cortical columns and often more than one cortical area. eLife Sciences Publications, Ltd 2020-11-06 /pmc/articles/PMC7647397/ /pubmed/33155981 http://dx.doi.org/10.7554/eLife.59841 Text en © 2020, Waters http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Neuroscience Waters, Jack Sources of widefield fluorescence from the brain |
title | Sources of widefield fluorescence from the brain |
title_full | Sources of widefield fluorescence from the brain |
title_fullStr | Sources of widefield fluorescence from the brain |
title_full_unstemmed | Sources of widefield fluorescence from the brain |
title_short | Sources of widefield fluorescence from the brain |
title_sort | sources of widefield fluorescence from the brain |
topic | Neuroscience |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7647397/ https://www.ncbi.nlm.nih.gov/pubmed/33155981 http://dx.doi.org/10.7554/eLife.59841 |
work_keys_str_mv | AT watersjack sourcesofwidefieldfluorescencefromthebrain |