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Scattered light fluorescence microscopy in three dimensions


Ghielmetti, Giulia; Aegerter, Christof M (2012). Scattered light fluorescence microscopy in three dimensions. Optics Express, 20(4):3744-3752.

Abstract

Recently, we have proposed a method to image fluorescent structures behind turbid layers at diffraction limited resolution using wave-front shaping and the memory effect. However, this was limited to a raster scanning of the wave-front shaped focus to a two dimensional plane. In applications, it can however be of great importance to be able to scan a three dimensional volume. Here we show that this can be implemented in the same setup. This is achieved by the addition of a parabolic phase shift to the shaped wave-front. Via the memory effect, this phase shift leads to a shift of the interference based focus in the z-direction, thus opening the possibility of three dimensional imaging using scattered light fluorescence microscopy. Here, we show an example of such a three dimensional image of fluorescent nano-beads taken behind a turbid layer more than 10 mean free paths thick. Finally, we discuss the differences of the scanning in the z-direction with that in the x–y plane and the corresponding possibilities and limitations of the technique.

Abstract

Recently, we have proposed a method to image fluorescent structures behind turbid layers at diffraction limited resolution using wave-front shaping and the memory effect. However, this was limited to a raster scanning of the wave-front shaped focus to a two dimensional plane. In applications, it can however be of great importance to be able to scan a three dimensional volume. Here we show that this can be implemented in the same setup. This is achieved by the addition of a parabolic phase shift to the shaped wave-front. Via the memory effect, this phase shift leads to a shift of the interference based focus in the z-direction, thus opening the possibility of three dimensional imaging using scattered light fluorescence microscopy. Here, we show an example of such a three dimensional image of fluorescent nano-beads taken behind a turbid layer more than 10 mean free paths thick. Finally, we discuss the differences of the scanning in the z-direction with that in the x–y plane and the corresponding possibilities and limitations of the technique.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:07 Faculty of Science > Physics Institute
Special Collections > SystemsX.ch
Special Collections > SystemsX.ch > Research, Technology and Development Projects > WingX
Dewey Decimal Classification:570 Life sciences; biology
530 Physics
Language:English
Date:2012
Deposited On:20 Feb 2013 14:05
Last Modified:05 Apr 2016 16:31
Publisher:Optical Society of America
ISSN:1094-4087
Publisher DOI:https://doi.org/10.1364/OE.20.003744

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