Building Accurate Intracellular Polarity Maps through Multiparametric Microscopy
| Autor: | Emilio Garcia-Fernandez, Pilar Herrero-Foncubierta, Angel Orte, M. Carmen Gonzalez-Garcia |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Fluorescence-lifetime imaging microscopy
Materials science QH301-705.5 Polarity (physics) solvatochromism 010402 general chemistry 01 natural sciences Biochemistry Genetics and Molecular Biology (miscellaneous) Fluorescence imaging Condensed Matter::Materials Science acridones fluorescence imaging Structural Biology Microscopy Calibration Fluorescence microscope Cellular microenvironment Technical Note Biology (General) lifetime fluorescence lifetime imaging microscopy (FLIM) 010405 organic chemistry Biosensing Solvatochromism Fluorescence Lifetime Imaging Microscopy (FLIM) cellular microenvironment 0104 chemical sciences biosensing Biological system Luminescence Lifetime Biotechnology Interpolation Acridones |
| Zdroj: | Methods and Protocols Digibug. Repositorio Institucional de la Universidad de Granada instname Methods and Protocols, Vol 3, Iss 78, p 78 (2020) |
| ISSN: | 2409-9279 2017-8565 |
| Popis: | M.C.G.-G. thanks MICIU/AEI for a predoctoral fellowship. We thank Juan M. Cuerva and Delia Miguel (University of Granada, Spain) for the synthesis of the acridone dyes, Rafael Salto and Maria D. Giron (University of Granada, Spain) for providing the cultured cells, and Amparo Navarro and Tomas Peña-Ruiz (University of Jaen, Spain) for computational studies of the acridone dyes. M.C.G.-G. thanks MICIU/AEI for a predoctoral fellowship. We thank Juan M. Cuerva and Delia Miguel (University of Granada, Spain) for the synthesis of the acridone dyes, Rafael Salto and Maria D. Giron (University of Granada, Spain) for providing the cultured cells, and Amparo Navarro and Tomas Peña-Ruiz (University of Jaen, Spain) for computational studies of the acridone dyes. 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. Spanish Ministerio de Ciencia e Innovación Agencia Estatal de Investigación and the European Regional Development Fund CTQ2017-85658-R. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|