Hyperspectral light sheet microscopy

Nature Communications
Wiebke JahrJan Huisken

Abstract

To study the development and interactions of cells and tissues, multiple fluorescent markers need to be imaged efficiently in a single living organism. Instead of acquiring individual colours sequentially with filters, we created a platform based on line-scanning light sheet microscopy to record the entire spectrum for each pixel in a three-dimensional volume. We evaluated data sets with varying spectral sampling and determined the optimal channel width to be around 5 nm. With the help of these data sets, we show that our setup outperforms filter-based approaches with regard to image quality and discrimination of fluorophores. By spectral unmixing we resolved overlapping fluorophores with up to nanometre resolution and removed autofluorescence in zebrafish and fruit fly embryos.

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Citations

Apr 6, 2016·Scientific Reports·Hoong-Ta Lim, Vadakke Matham Murukeshan
Apr 23, 2016·Scientific Reports·Jiamin WuQionghai Dai
Jul 28, 2016·Biophysical Journal·Zeno LavagninoDavid W Piston
Aug 23, 2016·Nature Methods·Chenchen PanAli Ertürk
Jan 10, 2017·Nature Methods·Francesco CutraleScott E Fraser
May 10, 2017·Journal of Biophotonics·Anna Siri LuthmanSarah E Bohndiek
Apr 1, 2017·Nature Methods·Rory M Power, Jan Huisken
Jul 28, 2017·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Jaroslav IchaCaren Norden
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Sep 7, 2018·Journal of Biophotonics·Lingling ChenShuangchen Ruan
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Methods Mentioned

BETA
confocal microscopy
chip
imaging technique
transgenic
light sheet

Software Mentioned

Fiji
MATLAB
LabVIEW
Andor Software Development Kit
Specim
LibreOffice

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