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

Petra Paiè; Roberto Memeo; Federico Sala; Michele Castriotta; Thomas Vaccari; Andrea Bassi; Roberto Osellame; Francesca Bragheri

doi:10.1117/12.2544873

17 February 2020 Dual-color on-chip light sheet microscopy of drosophila embryos

Petra Paiè, Roberto Memeo, Federico Sala, Michele Castriotta, Thomas Vaccari, Andrea Bassi, Roberto Osellame, Francesca Bragheri

Author Affiliations +

Proceedings Volume 11243, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XVIII; 112430K (2020) https://doi.org/10.1117/12.2544873
Event: SPIE BiOS, 2020, San Francisco, California, United States

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.

Conference Presentation

Citation Download Citation

Petra Paiè, Roberto Memeo, Federico Sala, Michele Castriotta, Thomas Vaccari, Andrea Bassi, Roberto Osellame, and Francesca Bragheri "Dual-color on-chip light sheet microscopy of drosophila embryos", Proc. SPIE 11243, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XVIII, 112430K (17 February 2020); https://doi.org/10.1117/12.2544873

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