Dynamic real-time subtraction of stray-light and background for multiphoton imaging.

Autor:	Fernández A; IQSE and Department of Soil and Crop Sciences, Texas A&M University, 4242 TAMU, College Station, TX 77843, USA.; Photonics Institute, TU Wien, Gusshausstrasse 27-29/387, 1040 Vienna, Austria.; Centro Regional Universitario de Coclé, Universidad de Panamá, Penonomé, Coclé, Panama.; alma.fernandez@tamu.edu., Straw A; Institute of Biology I and Bernstein Center Freiburg, University of Freiburg, Hauptstrasse 1, 79104 Freiburg, Germany., Distel M; St. Anna Children's Cancer Research Institute, Zimmermannplatz 10, 1090 Vienna, Austria., Leitgeb R; Center for Medical Physics and Biomedical Engineering, Medical University of Vienna, Währinger Gürtel 18-20/4L, 1090 Vienna, Austria., Baltuska A; Photonics Institute, TU Wien, Gusshausstrasse 27-29/387, 1040 Vienna, Austria., Verhoef AJ; IQSE and Department of Soil and Crop Sciences, Texas A&M University, 4242 TAMU, College Station, TX 77843, USA.; Photonics Institute, TU Wien, Gusshausstrasse 27-29/387, 1040 Vienna, Austria.; Centro Regional Universitario de Coclé, Universidad de Panamá, Penonomé, Coclé, Panama.; Center for Medical Physics and Biomedical Engineering, Medical University of Vienna, Währinger Gürtel 18-20/4L, 1090 Vienna, Austria.; aart.verhoef@tamu.edu.
Jazyk:	angličtina
Zdroj:	Biomedical optics express [Biomed Opt Express] 2020 Dec 14; Vol. 12 (1), pp. 288-302. Date of Electronic Publication: 2020 Dec 14 (Print Publication: 2021).
DOI:	10.1364/BOE.403255
Abstrakt:	We introduce a new approach to reduce uncorrelated background signals from fluorescence imaging data, using real-time subtraction of background light. This approach takes advantage of the short fluorescence lifetime of most popular fluorescent activity reporters, and the low duty-cycle of ultrafast lasers. By synchronizing excitation and recording, laser-induced multiphoton fluorescence can be discriminated from background light levels with each laser pulse. We demonstrate the ability of our method to - in real-time - remove image artifacts that in a conventional imaging setup lead to clipping of the signal. In other words, our method enables imaging under conditions that in a conventional setup would yield corrupted data from which no accurate information can be extracted. This is advantageous in experimental setups requiring additional light sources for applications such as optogenetic stimulation. Competing Interests: The authors declare no conflicts of interest. (Published by The Optical Society under the terms of the Creative Commons Attribution 4.0 License. Further distribution of this work must maintain attribution to the author(s) and the published article’s title, journal citation, and DOI.)
Databáze:	MEDLINE
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