Hyperspectral light sheet microscopy

Wiebke Jahr, Benjamin Schmid, Christopher Schmied, Florian O. Fahrbach and Jan Huisken ()
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Wiebke Jahr: Max Planck Institute of Molecular Cell Biology and Genetics
Benjamin Schmid: Max Planck Institute of Molecular Cell Biology and Genetics
Christopher Schmied: Max Planck Institute of Molecular Cell Biology and Genetics
Florian O. Fahrbach: Max Planck Institute of Molecular Cell Biology and Genetics
Jan Huisken: Max Planck Institute of Molecular Cell Biology and Genetics

Nature Communications, 2015, vol. 6, issue 1, 1-7

Abstract: 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.

Date: 2015
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DOI: 10.1038/ncomms8990

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