5/21/2023 0 Comments Fotonica lens![]() Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Jbio20200039-Movies.rarPDF document, 30 MB Averaging the FWHM of the intensity profile of different beads allows to determine the system lateral resolution The bead is acquired under double side blue illumination. Panel b and c shows the detail of a single nanobead with the corresponding profile characterization. In this image, left, right and double illumination for both blue and green excitation are reported. The error is calculated as the SD of the differences between the fit and the experimental points in absolute valuesįigure S6: Device characterization with nanobeads. In blue it is illustrated the intensity profile, while in red the gaussian fit, whose waist is also indicated in each graph (w). The analysis is performed for green and blue light and the measured resolution is about 1.055 and 0.993 μm, respectivelyįigure S5: Light sheets characterization performed analyzing the microscope images of the Rhodamine fluorescence. Table S1: Table with different nanobeads' spot dimensions used to retrieve the system lateral resolution. The dashed line is the outline of the laser machined structure, while the solid line is the profile of the etched structure the mismatch between the two is due to the acid attack along the unirradiated substrate (indicated by arrows) Panel (c) schematically shows the effect of the limited acid selectivity that must be compensated during the irradiation process to obtain the desired lens profile. Scale bar is 300 μmįigure S4: Microscope images of the top view of the integrated lens, before (panel a) and after (panel b) the etching process. ![]() ![]() The varied geometrical parameter is highlighted in red. The flow rate is 2 mL/h while the parameters h (a) and L (b) assume different values. ![]() (b) and (c) COMSOL simulations of an embryo flowing in the deviceįigure S3: Results of the COMSOL simulations for the flow of a Drosophila embryo inside the microchannel. Scale bar is 50 μmįigure S2: (a) Microscope image of the sample channel optimized for automatic particle rotation. Jbio20200039-Supinfo.pdfPDF document, 967.9 KBįigure S1 Shows different sections of an embryo oriented with its long axis parallel to the flow velocity profile, acquired during the sample movement through the light sheet. This work was made possible by the versatility of the used fabrication technique, femtosecond laser micromachining, which allows straightforward fabrication of both optical and fluidic components in glass substrates. In addition, we highlight the dual color investigation capabilities of this device, by processing two sample populations encoding different fluorescent proteins. In this regard, optical and fluidic optimization has been carried out to implement dual-sided illumination and automatic sample orientation. Indeed, the thickness and the high ellipticity of Drosophila embryos can degrade the image quality. The device has been fully optimized to address the challenges related to the specimens under investigation. This integrated device, constituted by both optical and microfluidic components, allows the automatic acquisition of a 3D stack of images for specimens diluted in a liquid suspension. We present a microscope on chip for automated imaging of Drosophila embryos by light sheet fluorescence microscopy.
0 Comments
Leave a Reply. |