Application of Spectral Phasor analysis to sodium microenvironments in myoblast progenitor cells.
Sodium ions (Na+) are key regulators of molecular events in many cellular processes, yet the dynamics of this ion remain poorly defined. Developing approaches to identify and characterise Na+ microenvironments will enable more detailed elucidation of the mechanisms of signal transduction. Here we re...
Main Authors: | , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Public Library of Science (PLoS)
2018-01-01
|
Series: | PLoS ONE |
Online Access: | http://europepmc.org/articles/PMC6209149?pdf=render |
_version_ | 1818194976468631552 |
---|---|
author | Hamid Sediqi Alex Wray Christopher Jones Mark Jones |
author_facet | Hamid Sediqi Alex Wray Christopher Jones Mark Jones |
author_sort | Hamid Sediqi |
collection | DOAJ |
description | Sodium ions (Na+) are key regulators of molecular events in many cellular processes, yet the dynamics of this ion remain poorly defined. Developing approaches to identify and characterise Na+ microenvironments will enable more detailed elucidation of the mechanisms of signal transduction. Here we report the application of Spectral Phasor analysis to the Na+ fluorophore, CoroNa Green, to identify and spatially map spectral emissions that report Na+ microenvironments. We use differentiating stem cells where Na+ fluxes were reported as an antecedent. Myoblast stem cells were induced to differentiate by serum starvation and then fixed at intervals between 0 and 40-minutes of differentiation prior to addition of CoroNa Green. The fluorescent intensity was insufficient to identify discrete Na+ microenvironments. However, using Spectral Phasor analysis we identified spectral shifts in CoroNa Green fluorescence which is related to the Na+ microenvironment. Further, spectral-heterogeneity appears to be contingent on the distance of Na+ from the nucleus in the early stages of differentiation. Spectral Phasor analysis of CoroNa Green in fixed stem cells demonstrates for the first time that CoroNa Green has unique spectral emissions depending on the nature of the Na+ environment in differentiating stem cells. Applying Spectral Phasor analysis to CoroNa Green in live stem cells is likely to further elucidate the role of Na+ microenvironments in the differentiation process. |
first_indexed | 2024-12-12T01:10:51Z |
format | Article |
id | doaj.art-00b4b8457d2b412d804ae53f179b46a2 |
institution | Directory Open Access Journal |
issn | 1932-6203 |
language | English |
last_indexed | 2024-12-12T01:10:51Z |
publishDate | 2018-01-01 |
publisher | Public Library of Science (PLoS) |
record_format | Article |
series | PLoS ONE |
spelling | doaj.art-00b4b8457d2b412d804ae53f179b46a22022-12-22T00:43:28ZengPublic Library of Science (PLoS)PLoS ONE1932-62032018-01-011310e020461110.1371/journal.pone.0204611Application of Spectral Phasor analysis to sodium microenvironments in myoblast progenitor cells.Hamid SediqiAlex WrayChristopher JonesMark JonesSodium ions (Na+) are key regulators of molecular events in many cellular processes, yet the dynamics of this ion remain poorly defined. Developing approaches to identify and characterise Na+ microenvironments will enable more detailed elucidation of the mechanisms of signal transduction. Here we report the application of Spectral Phasor analysis to the Na+ fluorophore, CoroNa Green, to identify and spatially map spectral emissions that report Na+ microenvironments. We use differentiating stem cells where Na+ fluxes were reported as an antecedent. Myoblast stem cells were induced to differentiate by serum starvation and then fixed at intervals between 0 and 40-minutes of differentiation prior to addition of CoroNa Green. The fluorescent intensity was insufficient to identify discrete Na+ microenvironments. However, using Spectral Phasor analysis we identified spectral shifts in CoroNa Green fluorescence which is related to the Na+ microenvironment. Further, spectral-heterogeneity appears to be contingent on the distance of Na+ from the nucleus in the early stages of differentiation. Spectral Phasor analysis of CoroNa Green in fixed stem cells demonstrates for the first time that CoroNa Green has unique spectral emissions depending on the nature of the Na+ environment in differentiating stem cells. Applying Spectral Phasor analysis to CoroNa Green in live stem cells is likely to further elucidate the role of Na+ microenvironments in the differentiation process.http://europepmc.org/articles/PMC6209149?pdf=render |
spellingShingle | Hamid Sediqi Alex Wray Christopher Jones Mark Jones Application of Spectral Phasor analysis to sodium microenvironments in myoblast progenitor cells. PLoS ONE |
title | Application of Spectral Phasor analysis to sodium microenvironments in myoblast progenitor cells. |
title_full | Application of Spectral Phasor analysis to sodium microenvironments in myoblast progenitor cells. |
title_fullStr | Application of Spectral Phasor analysis to sodium microenvironments in myoblast progenitor cells. |
title_full_unstemmed | Application of Spectral Phasor analysis to sodium microenvironments in myoblast progenitor cells. |
title_short | Application of Spectral Phasor analysis to sodium microenvironments in myoblast progenitor cells. |
title_sort | application of spectral phasor analysis to sodium microenvironments in myoblast progenitor cells |
url | http://europepmc.org/articles/PMC6209149?pdf=render |
work_keys_str_mv | AT hamidsediqi applicationofspectralphasoranalysistosodiummicroenvironmentsinmyoblastprogenitorcells AT alexwray applicationofspectralphasoranalysistosodiummicroenvironmentsinmyoblastprogenitorcells AT christopherjones applicationofspectralphasoranalysistosodiummicroenvironmentsinmyoblastprogenitorcells AT markjones applicationofspectralphasoranalysistosodiummicroenvironmentsinmyoblastprogenitorcells |