Back to Search
Start Over
Building Accurate Intracellular Polarity Maps through Multiparametric Microscopy.
- Source :
-
Methods and protocols [Methods Protoc] 2020 Nov 11; Vol. 3 (4). Date of Electronic Publication: 2020 Nov 11. - Publication Year :
- 2020
-
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.
Details
- Language :
- English
- ISSN :
- 2409-9279
- Volume :
- 3
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Methods and protocols
- Publication Type :
- Academic Journal
- Accession number :
- 33187290
- Full Text :
- https://doi.org/10.3390/mps3040078