We demonstrate the use of femtosecond laser micromachining for ablating macro-sized cavities in crystalline silicon. The method employed is laser milling in which the focused laser beam is raster scanned over the area to be removed. We report the achievement of very high volume ablation rates for the cavity of up to 8.48x106 μm3 s-1. To achieve such high rates, we make use of a high average power fiber laser source of 1030 nm wavelength and variable per pulse energy of up to 100 μJ. By carefully controlling the process variables such as pulse energy, repetition rate and scanner speed, the tradeoffs between micromachining quality and ablation rate are quantified. The developed process is applied on Siliconon-Insulator (SOI) wafers for improving performance of RF devices. By making use of laser removal and an additional step of selective silicon etch using XeF2, handler silicon is removed completely under RF circuits such as SP9T switch. The local removal of silicon under such circuits completely eliminates the losses and non-linearities caused by the coupling of RF signals to the semi-conducting substrate. Small-signal and large-signal RF measurements are performed before and after substrate removal to quantify the performance gain. The obtained performance after substrate removal is better than specialized RF-SOI substrates such as trap-rich SOI. This is of practical significance for next generation wireless technologies like 5G which operate at higher frequencies with stringent specifications. The proposed method is also potentially useful for fabricating membrane based devices in SOI technology such as pressure sensors.
Multiphoton microscopy is a cutting edge imaging modality leading to increasing advances in biology and also in the clinical field. To use it at its full potential and at the very heart of clinical practice, there have been several developments of fiber-based multiphoton microendoscopes. The application for those probes is now limited by few major restrictions, such as the difficulty to collect autofluorescence signals from tissues and cells theses being inherently weak (e.g. the ones from intracellular NADH or FAD metabolites). This limitation reduces the usefulness of microendoscopy in general, effectively restraining it to morphological imaging modality requiring staining of the tissues. Our aim is to go beyond this limitation, showing for the first time label-free cellular metabolism monitoring, in vivo in situ in real time.
The experimental setup is an upgrade of a recently published one (Ducourthial et.al, Scientific Reports, 2016) where femtosecond pulse fiber delivery is further optimized thank’s to a new transmissive-GRISM-based pulse stretcher permitting high energy throughput and wide bandwidth. This device allows fast sequential operation with two different excitation wavelengths for efficient two-photon excited NADH and FAD autofluorescence endoscopic detection (i.e. 860 nm for FAD and 760 nm for NADH), enabling cellular optical redox ratio quantification at 8 frames/s.
The obtained results on cell models in vitro and also on animal models in vivo (e.g. neurons of a living mouse) prove that we accurately assess the level of NADH and FAD at subcellular resolution through a 3-meters-long fiber with our miniaturized probe (O.D. =2.2 mm).
Fluorescence lifetime imaging microscopy (FLIM) represents a powerful tool for biological studies. Endoscopic FLIM applied to the intracellular native biomarker NADH and FAD represents a promising mean for in vivo in situ malignant tissue diagnosis in the medical field. Else, 2-photon-excited fluorescence (2PEF) provides increased 3D resolution and imaging depth. But very few demonstrations about 2PEF lifetime measurement through a fiber have been reported and none about endoscopic 2P-FLIM through a practical fiber length (< 3m).
Our group has recently demonstrated the possibility to efficiently deliver through a very long optical fiber the short and intense excitation pulses required for 2P-FLIM. Our goal is now to check that collecting fluorescence through the same endoscopic fiber does not deteriorate the lifetime measurement. Relying on the basis previously published in case of 1PEF by P. French and co-workers (J. Biophotonics, 2015), we have experimentally quantitatively evaluated the influence on the lifetime measurement of the fiber chromatic and intermodal dispersions. The main result is that the fiber contribution to the system impulse response function, even in the case of a 3-meter long double-clad optical fiber, does not hinder the separation between free and bound NADH states using FLIM. Related calibrations and measurements will be detailed. Ongoing experiments about the development of a 2P-FLIM endomicroscope on the basis of an previously reported 2P-endomicroscope (Ducourthial et al., Sc. Reports, 2015), used under various configurations (i.e. point measurement in the center of the 2P-endomicroscope image, averaged lifetime, binned endoscopic 2P-FLIM image), will be also presented.
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