Heterogeneity plays an important role in medicine and biology, which can be investigated by exploiting single cell analysis (SCA). Among SCA methods, imaging cytometry allows the analysis of individual 2D and 3D spatial features. Here we present a femtosecond laser fabricated optofluidic automated platform encompassing a thermo-optic phase shifter, cylindrical lenses and a microfluidic network to generate and shift a dual-color patterned light sheet within a microchannel where the samples of interest flow. The device can be used as add-on and can provide an acquisition rate of about 1 cell/second, or subnuclear resolution at the single cell level.
Integrated optical switches and modulators allow performing reconfigurability in integrated circuits, resulting as fundamental components in different fields ranging from optical communications to sensing and metrology. Among different methods, the thermo-optic effect has been successfully used to fabricate optical modulators by femtosecond laser micromachining (FLM) in glass substrates, proving high stability, no losses dependance but long switching time. In this work, we present an integrated optical switch realized by FLM with a switching time of less than 1 ms: which is about 1 order of magnitude faster than the other devices present in literature. This result has been achieved by carefully optimizing the geometry and the position of resistors and trenches near the waveguides through simulation and experimental validation. In addition, by means of an optimization of the applied voltage signal, we have demonstrated a further significant temporal improvement, measuring a switching time of less than 100 μs.
In this work we present a microscope on chip based on Light Sheet Fluorescence Microscopy, capable to automatically perform 3D and dual-color imaging of specimens diluted in a liquid suspension. A microfluidic channel is used for automatic sample delivery, while integrated optical components such as optical waveguides and lenses are used to illuminate the sample flowing in the channel. The device is fabricated by femtosecond laser micromachining in a glass substrate. Benefiting from the versatility of the fabrication technique we present two prototypes that have been optimized for different samples such as single cells and Drosophila embryos.
Lab on a Chip devices are compact and portable chips mainly constituted by a network of microfluidic channels. They aim at substituting bulk laboratory instrumentations, with the advantages of increasing the automation and the sensitivity of the analysis, reducing the costs and opening the possibility of performing measurements at the Point of Care. Among different Lab on Chips, optofluidic ones have the advantages of optical investigation, but the integration of optical and microfluidic components in a single substrate is very challenging from a technological point of view. A recent fabrication technique, known as femtosecond laser micromachining (FLM), has proven to be ideal for the realization of these devices, allowing the fabrication of the whole device in a single irradiation step. Here, we will present a platelet counter and a microscope on chip, that fully take advantage from the versatility of FLM. To succeed in these works a fundamental aspect to address is the capability to control the sample positioning in the microfluidic channel. A single particle per time should pass in the detection region to avoid the overlooking of specimen. Moreover, a precise control of the sample orientation and position in the channel cross section is needed for imaging. The 3D capabilities of FLM have been fundamental in the realization of advanced fluidic layouts capable of sample manipulation with no need of any additional external field. We have successfully proven red blood cells and platelets counting, as well as single cells, cellular spheroids and drosophila embryos 3D imaging.
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.
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