Evaluating the pharmacological response in fluorescence microscopy images: The Δm algorithm
| Autor: | Ana Isabel Gomez, M. Cruz, Juan F. López-Giménez |
|---|---|
| Přispěvatelé: | Universidad de Cantabria, Ministerio de Economía y Empresa (España) |
| Rok vydání: | 2019 |
| Předmět: |
Receptors
Opioid mu 02 engineering and technology Receptors G-Protein-Coupled Fluorescence Microscopy Antibiotics Microscopy Drug Discovery 0202 electrical engineering electronic engineering information engineering Fluorescence microscope Medicine and Health Sciences 0303 health sciences Analgesics Multidisciplinary Secretory Pathway Morphine Chemistry Antimicrobials Applied Mathematics Simulation and Modeling Light Microscopy Drugs Endocytosis Endocytic vesicle Cell Processes Doxycycline Physical Sciences Medicine 020201 artificial intelligence & image processing Cellular Structures and Organelles Algorithm Algorithms Research Article Endosome Imaging Techniques Science Endosomes Blob detection Research and Analysis Methods Microbiology 03 medical and health sciences Antimalarials Microbial Control Image Interpretation Computer-Assisted Fluorescence Imaging Humans Pain Management Computer Simulation Vesicles Transport Vesicles 030304 developmental biology Pharmacology Biology and Life Sciences Cell Biology Object detection Opioids Microscopy Fluorescence Temporal resolution Mathematics |
| Zdroj: | PLoS ONE 14(2): e0211330. UCrea Repositorio Abierto de la Universidad de Cantabria Universidad de Cantabria (UC) PLoS ONE Digital.CSIC. Repositorio Institucional del CSIC instname PLoS ONE, Vol 14, Iss 2, p e0211330 (2019) |
| Popis: | Current drug discovery procedures require fast and effective quantification of the pharmacological response evoked in living cells by agonist compounds. In the case of G-protein coupled receptors (GPCRs), the efficacy of a particular drug to initiate the endocytosis process is related to the formation of endocytic vesicles or endosomes and their subsequent internalisation within intracellular compartments that can be observed with high spatial and temporal resolution by fluorescence microscopy techniques. Recently, an algorithm has been proposed to evaluate the pharmacological response by estimating the number of endosomes per cell on time series of images. However, the algorithm was limited by the dependence on some manually set parameters and in some cases the quality of the image does not allow a reliable detection of the endosomes. Here we propose a simple, fast and automated image analysis method-the Delta m algorithm- to quantify a pharmacological response with data obtained from fluorescence microscopy experiments. This algorithm does not require individual object detection and computes the relative increment of the third order moment in fluorescence microscopy images after filtering with the Laplacian of Gaussian function. It was tested on simulations demonstrating its ability to discriminate different experimental situations according to the number and the fluorescence signal intensity of the simulated endosomes. Finally and in order to validate this methodology with real data, the algorithm was applied to several time-course experiments based on the endocytosis of the mu opioid receptor (MOP) initiated by different agonist compounds. Each drug displayed a different Am sigmoid time-response curve and statistically significant differences were observed among drugs in terms of efficacy and kinetic parameters. The authors acknowledge financial support from the Spanish Project AYA2015-66357-R 288 (MINECO/FEDER). |
| Databáze: | OpenAIRE |
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