Research ExplorerPublicationsA Software Tool for High-Throughput Real-Time Measurement of...

A Software Tool for High-Throughput Real-Time Measurement of Intensity-Based Ratio-Metric FRET

Standard

A Software Tool for High-Throughput Real-Time Measurement of Intensity-Based Ratio-Metric FRET. / Ramuz, Masoud; Hasan, Alveera; Gruscheski, Lena; Diakonov, Ivan; Pavlaki, Nikoleta; Nikolaev, Viacheslav O; Harding, Sian; Dunsby, Chris; Gorelik, Julia.

In: CELLS-BASEL, Vol. 8, No. 12, 29.11.2019.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Ramuz, M, Hasan, A, Gruscheski, L, Diakonov, I, Pavlaki, N, Nikolaev, VO, Harding, S, Dunsby, C & Gorelik, J 2019, 'A Software Tool for High-Throughput Real-Time Measurement of Intensity-Based Ratio-Metric FRET', CELLS-BASEL, vol. 8, no. 12. https://doi.org/10.3390/cells8121541

APA

Ramuz, M., Hasan, A., Gruscheski, L., Diakonov, I., Pavlaki, N., Nikolaev, V. O., Harding, S., Dunsby, C., & Gorelik, J. (2019). A Software Tool for High-Throughput Real-Time Measurement of Intensity-Based Ratio-Metric FRET. CELLS-BASEL, 8(12). https://doi.org/10.3390/cells8121541

Vancouver

Ramuz M, Hasan A, Gruscheski L, Diakonov I, Pavlaki N, Nikolaev VO et al. A Software Tool for High-Throughput Real-Time Measurement of Intensity-Based Ratio-Metric FRET. CELLS-BASEL. 2019 Nov 29;8(12). https://doi.org/10.3390/cells8121541

Bibtex

@article{1f9d49130f054df19e42abffb6fef824,
title = "A Software Tool for High-Throughput Real-Time Measurement of Intensity-Based Ratio-Metric FRET",
abstract = "F{\"o}rster resonance energy transfer (FRET) is increasingly used for non-invasive measurement of fluorescently tagged molecules in live cells. In this study, we have developed a freely available software tool MultiFRET, which, together with the use of a motorised microscope stage, allows multiple single cells to be studied in one experiment. MultiFRET is a Java plugin for Micro-Manager software, which provides real-time calculations of ratio-metric signals during acquisition and can simultaneously record from multiple cells in the same experiment. It can also make other custom-determined live calculations that can be easily exported to Excel at the end of the experiment. It is flexible and can work with multiple spectral acquisition channels. We validated this software by comparing the output of MultiFRET to that of a previously established and well-documented method for live ratio-metric FRET experiments and found no significant difference between the data produced with the use of the new MultiFRET and other methods. In this validation, we used several cAMP FRET sensors and cell models: i) isolated adult cardiomyocytes from transgenic mice expressing the cytosolic epac1-camps and targeted pmEpac1 and Epac1-PLN sensors, ii) isolated neonatal mouse cardiomyocytes transfected with the AKAP79-CUTie sensor, and iii) human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) transfected with the Epac-SH74 sensor. The MultiFRET plugin is an open source freely available package that can be used in a wide area of live cell imaging when live ratio-metric calculations are required.",
author = "Masoud Ramuz and Alveera Hasan and Lena Gruscheski and Ivan Diakonov and Nikoleta Pavlaki and Nikolaev, {Viacheslav O} and Sian Harding and Chris Dunsby and Julia Gorelik",
year = "2019",
month = nov,
day = "29",
doi = "10.3390/cells8121541",
language = "English",
volume = "8",
journal = "CELLS-BASEL",
issn = "2073-4409",
publisher = "MDPI Multidisciplinary Digital Publishing Institute",
number = "12",

}

RIS

TY - JOUR

T1 - A Software Tool for High-Throughput Real-Time Measurement of Intensity-Based Ratio-Metric FRET

AU - Ramuz, Masoud

AU - Hasan, Alveera

AU - Gruscheski, Lena

AU - Diakonov, Ivan

AU - Pavlaki, Nikoleta

AU - Nikolaev, Viacheslav O

AU - Harding, Sian

AU - Dunsby, Chris

AU - Gorelik, Julia

PY - 2019/11/29

Y1 - 2019/11/29

N2 - Förster resonance energy transfer (FRET) is increasingly used for non-invasive measurement of fluorescently tagged molecules in live cells. In this study, we have developed a freely available software tool MultiFRET, which, together with the use of a motorised microscope stage, allows multiple single cells to be studied in one experiment. MultiFRET is a Java plugin for Micro-Manager software, which provides real-time calculations of ratio-metric signals during acquisition and can simultaneously record from multiple cells in the same experiment. It can also make other custom-determined live calculations that can be easily exported to Excel at the end of the experiment. It is flexible and can work with multiple spectral acquisition channels. We validated this software by comparing the output of MultiFRET to that of a previously established and well-documented method for live ratio-metric FRET experiments and found no significant difference between the data produced with the use of the new MultiFRET and other methods. In this validation, we used several cAMP FRET sensors and cell models: i) isolated adult cardiomyocytes from transgenic mice expressing the cytosolic epac1-camps and targeted pmEpac1 and Epac1-PLN sensors, ii) isolated neonatal mouse cardiomyocytes transfected with the AKAP79-CUTie sensor, and iii) human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) transfected with the Epac-SH74 sensor. The MultiFRET plugin is an open source freely available package that can be used in a wide area of live cell imaging when live ratio-metric calculations are required.

AB - Förster resonance energy transfer (FRET) is increasingly used for non-invasive measurement of fluorescently tagged molecules in live cells. In this study, we have developed a freely available software tool MultiFRET, which, together with the use of a motorised microscope stage, allows multiple single cells to be studied in one experiment. MultiFRET is a Java plugin for Micro-Manager software, which provides real-time calculations of ratio-metric signals during acquisition and can simultaneously record from multiple cells in the same experiment. It can also make other custom-determined live calculations that can be easily exported to Excel at the end of the experiment. It is flexible and can work with multiple spectral acquisition channels. We validated this software by comparing the output of MultiFRET to that of a previously established and well-documented method for live ratio-metric FRET experiments and found no significant difference between the data produced with the use of the new MultiFRET and other methods. In this validation, we used several cAMP FRET sensors and cell models: i) isolated adult cardiomyocytes from transgenic mice expressing the cytosolic epac1-camps and targeted pmEpac1 and Epac1-PLN sensors, ii) isolated neonatal mouse cardiomyocytes transfected with the AKAP79-CUTie sensor, and iii) human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) transfected with the Epac-SH74 sensor. The MultiFRET plugin is an open source freely available package that can be used in a wide area of live cell imaging when live ratio-metric calculations are required.

U2 - 10.3390/cells8121541

DO - 10.3390/cells8121541

M3 - SCORING: Journal article

C2 - 31795419

VL - 8

JO - CELLS-BASEL

JF - CELLS-BASEL

SN - 2073-4409

IS - 12

ER -