Light-sheet microscopy with length-adaptive Bessel beams

Biomedical Optics Express
Tobias Meinert, Alexander Rohrbach

Abstract

In light-sheet microscopy, a confined layer in the focal plane of the detection objective is illuminated from the side. The illumination light-sheet usually has a constant beam length independent of the shape of the biological object. Since the thickness and the length of the illumination light-sheet are coupled, a tradeoff between resolution, contrast and field of view has to be accepted. Here we show that scanned Bessel beams enable object adapted tailoring of the light-sheet defined by its beam length and position. The individual beam parameters are obtained from automatic object shape estimation by low-power laser light scattered at the object. Using Arabidopsis root tips, cell clusters and zebrafish tails, we demonstrate that Bessel beam light-sheet tailoring leads to a 50% increase in image contrast and a 50% reduction in photobleaching. Light-sheet tailoring requires only binary amplitude modulation, therefore allowing a real time illumination adaptation with little technical effort in the future.

References

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Citations

Feb 8, 2020·Scientific Reports·Gustavo de MedeirosLars Hufnagel
May 22, 2021·Nature Communications·Sota TakanezawaTakeshi Imamura

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