Building Accurate Intracellular Polarity Maps through Multiparametric Microscopy

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ID: 266453
2020
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Abstract
The precise knowledge of intracellular polarity, a physiological parameter that involves complex and intertwined intracellular mechanisms, may be relevant in the study of important diseases like cancer or Alzheimer’s. In this technical note, we illustrate our recently developed, accurate method for obtaining intracellular polarity maps employing potent fluorescence microscopy techniques. Our method is based on the selection of appropriate luminescent probes, in which several emission properties vary with microenvironment polarity, specifically spectral shifts and luminescence lifetime. A multilinear calibration is performed, correlating polarity vs. spectral shift vs. luminescence lifetime, to generate a powerful and error-free 3D space for reliable interpolation of microscopy data. Multidimensional luminescence microscopy is then used to obtain simultaneously spectral shift and luminescence lifetime images, which are then interpolated in the 3D calibration space, resulting in accurate, quantitative polarity maps.
Reference Key
gonzalez-garcia2020methodsbuilding Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors M. Carmen Gonzalez-Garcia;Pilar Herrero-Foncubierta;Emilio Garcia-Fernandez;Angel Orte;Gonzalez-Garcia, M. Carmen;Herrero-Foncubierta, Pilar;Garcia-Fernandez, Emilio;Orte, Angel;
Journal methods and protocols
Year 2020
DOI
10.3390/mps3040078
URL
https://www.mdpi.com/2409-9279/3/4/78#citedby
Keywords
solvatochromism fluorescence imaging biosensing lifetime cellular microenvironment fluorescence lifetime imaging microscopy (flim) acridones

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