Fluorescence Microscopy-Based Quantitation of GLUT4 Translocation: High Throughput or High Content?
Abstract
:1. Introduction
2. Results
2.1. Quantitation of GLUT4 Translocation by Objective-type Scanning TIRF Microscopy
2.2. Quantitation of GLUT4 Translocation Using A Prism-Type TIR Reader
2.3. Influence of Autofluorescence of Tested Compounds on the Quantitation of GLUT4 Translocation
2.4. Quantitation of GLUT4 Translocation by Confocal Microscopy of Giant Plasma Membrane Vesicles (GPMVs)
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Cell Culture and Transfection
4.3. Objective-Type TIRF Microscopy
4.4. Prism-Type TIR Multiwell Reader
4.5. Fluorescence Spectra for Autofluorescence and Quenching Properties of Plant Extracts
4.6. Giant Plasma Membrane Vesicle (GPMV) Formation and Confocal Imaging
4.7. Data Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
GLUT4 | Glucose transporter 4 |
GPMV | Giant plasma membrane vesicle |
T2DM | Type 2 diabetes mellitus |
TIRF | Total internal reflection fluorescence |
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Stadlbauer, V.; Lanzerstorfer, P.; Neuhauser, C.; Weber, F.; Stübl, F.; Weber, P.; Wagner, M.; Plochberger, B.; Wieser, S.; Schneckenburger, H.; et al. Fluorescence Microscopy-Based Quantitation of GLUT4 Translocation: High Throughput or High Content? Int. J. Mol. Sci. 2020, 21, 7964. https://doi.org/10.3390/ijms21217964
Stadlbauer V, Lanzerstorfer P, Neuhauser C, Weber F, Stübl F, Weber P, Wagner M, Plochberger B, Wieser S, Schneckenburger H, et al. Fluorescence Microscopy-Based Quantitation of GLUT4 Translocation: High Throughput or High Content? International Journal of Molecular Sciences. 2020; 21(21):7964. https://doi.org/10.3390/ijms21217964
Chicago/Turabian StyleStadlbauer, Verena, Peter Lanzerstorfer, Cathrina Neuhauser, Florian Weber, Flora Stübl, Petra Weber, Michael Wagner, Birgit Plochberger, Stefan Wieser, Herbert Schneckenburger, and et al. 2020. "Fluorescence Microscopy-Based Quantitation of GLUT4 Translocation: High Throughput or High Content?" International Journal of Molecular Sciences 21, no. 21: 7964. https://doi.org/10.3390/ijms21217964