Fraping: A computational tool for detecting slight differences in fluorescence recovery after photobleaching (FRAP) data for actin polymerization analysis.
F-actin
FRAP
LifeAct
neurites
neuron growth
neurons
simulation
software analysis
Journal
Microscopy research and technique
ISSN: 1097-0029
Titre abrégé: Microsc Res Tech
Pays: United States
ID NLM: 9203012
Informations de publication
Date de publication:
01 Mar 2024
01 Mar 2024
Historique:
revised:
13
02
2024
received:
30
08
2023
accepted:
14
02
2024
medline:
1
3
2024
pubmed:
1
3
2024
entrez:
1
3
2024
Statut:
aheadofprint
Résumé
Fluorescence recovery after photobleaching (FRAP) is a laser method of light microscopy to evaluate the rapid movement of fluorescent molecules. To have a more reliable approach to analyze data from FRAP, we designed Fraping, a free access R library to data analysis obtained from FRAP. Unlike other programs, Fraping has a new form of analyzing curves of FRAP using statistical analysis based on the average curve difference. To evaluate our library, we analyzed the differences of actin polymerization in real time between dendrites and secondary neurites of cultured neuron transfected with LifeAct to track F-actin changes of neurites. We found that Fraping provided greater sensitivity than the conventional model using mobile fraction analysis. Likewise, this approach allowed us to normalize the fluorescence to the size area of interest and adjust data curves choosing the best parametric model. In addition, this library was supplemented with data simulation to have a more significant enrichment for the analysis behavior. We concluded that Fraping is a method that reduces bias when analyzing two data groups as compared with the conventional methods. This method also allows the users to choose a more suitable analysis approach according to their requirements. RESEARCH HIGHLIGHTS: Fraping is a new programming tool to analyze FRAP data to normalize fluorescence recovery curves. The conventional method uses one-point analysis, and the new one compares all the points to define the similarity of the fluorescence recovery.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Consejo Nacional de Humanidades, Ciencia y Tecnología
ID : 285184
Organisme : Dirección General de Asuntos del Personal Académico, UNAM
ID : IN212019
Organisme : Dirección General de Asuntos del Personal Académico, UNAM
ID : IN216422
Informations de copyright
© 2024 The Authors. Microscopy Research and Technique published by Wiley Periodicals LLC.
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