4.4 Article

Novel algorithms for improved detection and analysis of fluorescent signal fluctuations

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SPRINGER HEIDELBERG
DOI: 10.1007/s00424-023-02855-3

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Calcium signal analysis; ROI detection; Background correction; Transient classification; Interactive user interface; Glial calcium signals; Neuronal SBFI imaging

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Fluorescent dyes and genetically encoded fluorescence indicators are commonly used tools for visualizing concentration changes of specific ions and messenger molecules. Two algorithms (PBasE and CoRoDe) are presented in this study to accurately estimate baseline and automate the detection and segmentation of fluorescence fluctuations.
Fluorescent dyes and genetically encoded fluorescence indicators (GEFI) are common tools for visualizing concentration changes of specific ions and messenger molecules during intra- as well as intercellular communication. Using advanced imaging technologies, fluorescence indicators are a prerequisite for the analysis of physiological molecular signaling. Automated detection and analysis of fluorescence signals require to overcome several challenges, including correct estimation of fluorescence fluctuations at basal concentrations of messenger molecules, detection, and extraction of events themselves as well as proper segmentation of neighboring events. Moreover, event detection algorithms need to be sensitive enough to accurately capture localized and low amplitude events exhibiting a limited spatial extent. Here, we present two algorithms (PBasE and CoRoDe) for accurate baseline estimation and automated detection and segmentation of fluorescence fluctuations.

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