Increased Accuracy to c-Fos-Positive Neuron Counting

There is not a described method to count the core label of c-Fos-positive neurons, avoiding false-positive and false-negative results. The aim of this manuscript is to provide guidelines for a secure and accurate method to calculate a threshold to select which core of c-Fos-positive neurons marked b...

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Autores principales:	Wellington José da Silva, José Rodrigo Santos Silva, Jullyana de Souza Siqueira Quintans, Waldecy de Lucca Junior
Formato:	article
Lenguaje:	EN
Publicado:	Hindawi Limited 2021
Materias:	Medicine R
Acceso en línea:	https://doaj.org/article/18fa74f6f38c4ad5808fb5e2bcf42d80
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spelling	oai:doaj.org-article:18fa74f6f38c4ad5808fb5e2bcf42d802021-11-22T01:09:40ZIncreased Accuracy to c-Fos-Positive Neuron Counting2314-614110.1155/2021/3060983https://doaj.org/article/18fa74f6f38c4ad5808fb5e2bcf42d802021-01-01T00:00:00Zhttp://dx.doi.org/10.1155/2021/3060983https://doaj.org/toc/2314-6141There is not a described method to count the core label of c-Fos-positive neurons, avoiding false-positive and false-negative results. The aim of this manuscript is to provide guidelines for a secure and accurate method to calculate a threshold to select which core of c-Fos-positive neurons marked by immunofluorescence has to be scored. A background percentage was calculated by dividing the intensity value (0 to 255) of the core of c-Fos-positive neurons by its surrounding background from the 8-bit images obtained in a previous study. Using the background percentage from 20% up to 98%, raising 2% once for each score, as threshold to choose which core has to be counted, a script was written for the R program to count the number of the c-Fos-positive neurons and the comparison between control and experimental groups. The differences of the average number of the core counted c-Fos-positive neurons between control and experimental groups, at all thresholds studied, showed a rising value related to an increase of the background percentage threshold as well as a decrease of its p value related to an increase of the threshold of background percentage. For the smallest thresholds (high intensity of label), the differences between groups are suppressed (false negative). However, for the biggest thresholds (nonspecific label), these differences are always the same (false positive). Therefore, to avoid the false-negative and the false-positive values, it was chosen as the threshold of 62% the inflection point of the linear regression, which is equally different from the biggest and smallest values of the differences between groups.Wellington José da SilvaJosé Rodrigo Santos SilvaJullyana de Souza Siqueira QuintansWaldecy de Lucca JuniorHindawi LimitedarticleMedicineRENBioMed Research International, Vol 2021 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R
spellingShingle	Medicine R Wellington José da Silva José Rodrigo Santos Silva Jullyana de Souza Siqueira Quintans Waldecy de Lucca Junior Increased Accuracy to c-Fos-Positive Neuron Counting
description	There is not a described method to count the core label of c-Fos-positive neurons, avoiding false-positive and false-negative results. The aim of this manuscript is to provide guidelines for a secure and accurate method to calculate a threshold to select which core of c-Fos-positive neurons marked by immunofluorescence has to be scored. A background percentage was calculated by dividing the intensity value (0 to 255) of the core of c-Fos-positive neurons by its surrounding background from the 8-bit images obtained in a previous study. Using the background percentage from 20% up to 98%, raising 2% once for each score, as threshold to choose which core has to be counted, a script was written for the R program to count the number of the c-Fos-positive neurons and the comparison between control and experimental groups. The differences of the average number of the core counted c-Fos-positive neurons between control and experimental groups, at all thresholds studied, showed a rising value related to an increase of the background percentage threshold as well as a decrease of its p value related to an increase of the threshold of background percentage. For the smallest thresholds (high intensity of label), the differences between groups are suppressed (false negative). However, for the biggest thresholds (nonspecific label), these differences are always the same (false positive). Therefore, to avoid the false-negative and the false-positive values, it was chosen as the threshold of 62% the inflection point of the linear regression, which is equally different from the biggest and smallest values of the differences between groups.
format	article
author	Wellington José da Silva José Rodrigo Santos Silva Jullyana de Souza Siqueira Quintans Waldecy de Lucca Junior
author_facet	Wellington José da Silva José Rodrigo Santos Silva Jullyana de Souza Siqueira Quintans Waldecy de Lucca Junior
author_sort	Wellington José da Silva
title	Increased Accuracy to c-Fos-Positive Neuron Counting
title_short	Increased Accuracy to c-Fos-Positive Neuron Counting
title_full	Increased Accuracy to c-Fos-Positive Neuron Counting
title_fullStr	Increased Accuracy to c-Fos-Positive Neuron Counting
title_full_unstemmed	Increased Accuracy to c-Fos-Positive Neuron Counting
title_sort	increased accuracy to c-fos-positive neuron counting
publisher	Hindawi Limited
publishDate	2021
url	https://doaj.org/article/18fa74f6f38c4ad5808fb5e2bcf42d80
work_keys_str_mv	AT wellingtonjosedasilva increasedaccuracytocfospositiveneuroncounting AT joserodrigosantossilva increasedaccuracytocfospositiveneuroncounting AT jullyanadesouzasiqueiraquintans increasedaccuracytocfospositiveneuroncounting AT waldecydeluccajunior increasedaccuracytocfospositiveneuroncounting
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Increased Accuracy to c-Fos-Positive Neuron Counting

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