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Proceedings Paper

Dual-color on-chip light sheet microscopy of drosophila embryos
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Paper Abstract

Drosophila Melanogaster is a sample of high biological interest that is being widely used as biological model, due to the relatively short life cycle, short genome and ease in culturing. In this work we present a microscope on chip capable of processing Drosophila embryos to obtain three dimensional fluorescent images at high throughput. This device, based on light sheet microscopy, uses a plane of light intercepting the sample channel to optically and noninvasively section the embryos while flowing. This permits to automatically acquire for each sample the stack of images necessary for the subsequent 3D reconstruction with no need of any manual sample positioning and alignment. The whole chip is fabricated in a glass substrate by femtosecond laser micromachining. The device has been optimized for the specific morphology of the sample. Indeed, the highly elliptical shape of the embryos (about 100 x 500 μm2) might affect the image quality degrading both the vertical and the axial resolution of the system. To overcome this issue, we have first optimized the layout of the fluidic channel to precisely control the sample orientation by means of hydrodynamic forces. Thereafter, we have optimized the properties of the optical circuit, to realize two opposite light sheets impinging on the sample, perfectly overlapped, with a high signal to noise ratio. With these actions, we have been able to obtain high quality Drosophila reconstruction.

Paper Details

Date Published: 17 February 2020
PDF: 6 pages
Proc. SPIE 11243, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XVIII, 112430K (17 February 2020); doi: 10.1117/12.2544873
Show Author Affiliations
Petra Paiè, CNR-Istituto di Fotonica e Nanotecnologie (Italy)
Roberto Memeo, Politecnico di Milano (Italy)
Federico Sala, CNR-Istituto di Fotonica e Nanotecnologie (Italy)
Politecnico di Milano (Italy)
Michele Castriotta, Politecnico di Milano (Italy)
Thomas Vaccari, Univ. degli Studi di Milano (Italy)
Andrea Bassi, Politecnico di Milano (Italy)
Roberto Osellame, CNR-Istituto di Fotonica e Nanotecnologie (Italy)
Politecnico di Milano (Italy)
Francesca Bragheri, CNR-Istituto di Fotonica e Nanotecnologie (Italy)

Published in SPIE Proceedings Vol. 11243:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XVIII
Daniel L. Farkas; Attila Tarnok, Editor(s)

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