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Accurate Rapid Lifetime Determination on Time-Gated FLIM Microscopy with Optical Sectioning
Joint Authors
Silva, Susana F.
Domingues, José Paulo
Morgado, António Miguel
Source
Journal of Healthcare Engineering
Issue
Vol. 2018, Issue 2018 (31 Dec. 2018), pp.1-7, 7 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2018-01-16
Country of Publication
Egypt
No. of Pages
7
Main Subjects
Abstract EN
Time-gated fluorescence lifetime imaging microscopy (FLIM) is a powerful technique to assess the biochemistry of cells and tissues.
When applied to living thick samples, it is hampered by the lack of optical sectioning and the need of acquiring many images for an accurate measurement of fluorescence lifetimes.
Here, we report on the use of processing techniques to overcome these limitations, minimizing the acquisition time, while providing optical sectioning.
We evaluated the application of the HiLo and the rapid lifetime determination (RLD) techniques for accurate measurement of fluorescence lifetimes with optical sectioning.
HiLo provides optical sectioning by combining the high-frequency content from a standard image, obtained with uniform illumination, with the low-frequency content of a second image, acquired using structured illumination.
Our results show that HiLo produces optical sectioning on thick samples without degrading the accuracy of the measured lifetimes.
We also show that instrument response function (IRF) deconvolution can be applied with the RLD technique on HiLo images, improving greatly the accuracy of the measured lifetimes.
These results open the possibility of using the RLD technique with pulsed diode laser sources to determine accurately fluorescence lifetimes in the subnanosecond range on thick multilayer samples, providing that offline processing is allowed.
American Psychological Association (APA)
Silva, Susana F.& Domingues, José Paulo& Morgado, António Miguel. 2018. Accurate Rapid Lifetime Determination on Time-Gated FLIM Microscopy with Optical Sectioning. Journal of Healthcare Engineering،Vol. 2018, no. 2018, pp.1-7.
https://search.emarefa.net/detail/BIM-1186904
Modern Language Association (MLA)
Silva, Susana F.…[et al.]. Accurate Rapid Lifetime Determination on Time-Gated FLIM Microscopy with Optical Sectioning. Journal of Healthcare Engineering No. 2018 (2018), pp.1-7.
https://search.emarefa.net/detail/BIM-1186904
American Medical Association (AMA)
Silva, Susana F.& Domingues, José Paulo& Morgado, António Miguel. Accurate Rapid Lifetime Determination on Time-Gated FLIM Microscopy with Optical Sectioning. Journal of Healthcare Engineering. 2018. Vol. 2018, no. 2018, pp.1-7.
https://search.emarefa.net/detail/BIM-1186904
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1186904