Publicação
SIGAA: signaling automated analysis: a new tool for Ca2+ signaling quantification using ratiometric Ca2+ dyes
| Resumo: | Astrocytes are non-neural cells, restricted to the brain and spinal cord, whose functions and morphology depend on their location. Astrocyte–astrocyte and astrocyte–neuron interactions occur through cytoplasmic Ca2+ level changes that are assessed to determine cell function and response (i.e., drug testing). The evaluation of alterations in intracellular Ca2+ levels primarily relies on fluorescence imaging techniques, performed through video recording of cells incubated with Ca2+-sensitive dyes. By observing ion concentration shifts over time in a delimited region of interest (ROI) encompassing a single cell, it is possible to draw conclusions on cell responses to specific stimuli. Our work describes a tool named SIGAA—signaling automated analysis, for astrocyte ROI-based fluorescent imaging. This tool is specifically tailored for two wavelengths excited dyes by using two inputs of Ca2+ signaling recorded frames/videos and outputting a set of features relevant to the experiment’s conclusions and cell characterization. SIGAA performs automatic drift correction for the two recorded videos with a template matching algorithm, followed by astrocyte identification (ROI) using morphological reconstruction techniques. Subsequently, SIGAA extracts intracellular Ca2+ evolution functions for all identified ROIs detects function transients, and estimates a set of features for each signal. These features closely resemble those obtained through traditional methods and software used thus far. SIGAA is a new fully automated tool, which can speed up hour-long studies and analysis to a few minutes, showing reliable results as the validity tests indicate. |
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| Autores principais: | Lopes, Rafael Faria |
| Outros Autores: | Gonçalves-Ribeiro, Joana; Sebastião, Ana M.; Meneses, Carlos; Vaz, Sandra H. |
| Assunto: | Astrocyte Central nervous system Calcium (Ca2+) ion Automation Image processing Video drift correction Template matching Automatic detection Morphological reconstruction |
| Ano: | 2023 |
| País: | Portugal |
| Tipo de documento: | artigo original |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Instituto Politécnico de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório Científico do Instituto Politécnico de Lisboa |
| Resumo: | Astrocytes are non-neural cells, restricted to the brain and spinal cord, whose functions and morphology depend on their location. Astrocyte–astrocyte and astrocyte–neuron interactions occur through cytoplasmic Ca2+ level changes that are assessed to determine cell function and response (i.e., drug testing). The evaluation of alterations in intracellular Ca2+ levels primarily relies on fluorescence imaging techniques, performed through video recording of cells incubated with Ca2+-sensitive dyes. By observing ion concentration shifts over time in a delimited region of interest (ROI) encompassing a single cell, it is possible to draw conclusions on cell responses to specific stimuli. Our work describes a tool named SIGAA—signaling automated analysis, for astrocyte ROI-based fluorescent imaging. This tool is specifically tailored for two wavelengths excited dyes by using two inputs of Ca2+ signaling recorded frames/videos and outputting a set of features relevant to the experiment’s conclusions and cell characterization. SIGAA performs automatic drift correction for the two recorded videos with a template matching algorithm, followed by astrocyte identification (ROI) using morphological reconstruction techniques. Subsequently, SIGAA extracts intracellular Ca2+ evolution functions for all identified ROIs detects function transients, and estimates a set of features for each signal. These features closely resemble those obtained through traditional methods and software used thus far. SIGAA is a new fully automated tool, which can speed up hour-long studies and analysis to a few minutes, showing reliable results as the validity tests indicate. |
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