Claire Lefort

Pulse compression and fiber delivery of 45 fs Fourier transform limited pulses at 830 nm

A specific scheme is used for fiber delivery of ultrashort pulses using conventional elements. Starting from a standard femtosecond Ti:Al(2)O(3) oscillator (150 fs @ 830 nm), perfectly compressed ultrashort pulses with a duration of 45 fs are produced at the output of a standard two meter long single-mode fiber. The setup allows compensating independently and simultaneously second and third orders of chromatic dispersion as well as management of self-phase modulation in the fiber. It includes an optimized dispersion compensation line made of the assembly of diffraction gratings and prisms. The unsurpassed performances of the device are experimentally and numerically highlighted. Fiber delivery of sub-30 fs multinanojoule pulses is discussed.

Ultrashort pulse fiber delivery with optimized dispersion control by reflection grisms at 800 nm

We experimentally demonstrate a compact and efficient arrangement for fiber delivery of sub-30 fs energetic light pulses at 800 nm. Pulses coming from a broadband Ti:Sapphire oscillator are negatively pre-chirped by a grism-pair stretcher that allows for the control of second and third orders of dispersion. At the direct exit of a 2.7-m long large mode area (LMA) photonic crystal fiber 1-nJ pulses are temporally compressed to 29 fs producing close to 30 kW of peak power. The tunability of the device is studied. Comparison between LMA fibers and standard SMF fibers is also discussed.

Development of a nonlinear fiber-optic spectrometer for human lung tissue exploration

Several major lung pathologies are characterized by early modifications of the extracellular matrix (ECM) fibrillar collagen and elastin network. We report here the development of a nonlinear fiber-optic spectrometer, compatible with an endoscopic use, primarily intended for the recording of second-harmonic generation (SHG) signal of collagen and two-photon excited fluorescence (2PEF) of both collagen and elastin. Fiber dispersion is accurately compensated by the use of a specific grism-pair stretcher, allowing laser pulse temporal width around 70 fs and excitation wavelength tunability from 790 to 900 nm. This spectrometer was used to investigate the excitation wavelength dependence (from 800 to 870 nm) of SHG and 2PEF spectra originating from ex vivo human lung tissue samples. The results were compared with spectral responses of collagen gel and elastin powder reference samples and also with data obtained using standard nonlinear microspectroscopy. The excitation-wavelength-tunable nonlinear fiber-optic spectrometer presented in this study allows performing nonlinear spectroscopy of human lung tissue ECM through the elastin 2PEF and the collagen SHG signals. This work opens the way to tunable excitation nonlinear endomicroscopy based on both distal scanning of a single optical fiber and proximal scanning of a fiber-optic bundle.

Characterization, comparison, and choice of a commercial double-clad fiber for nonlinear endomicroscopy

Abstract. Several endomicroscope prototypes for nonlinear optical imaging were developed in the last decade for in situ analysis of tissue with cellular resolution by using short infrared light pulses. Fourier-transform-limited pulses at the tissue site are necessary for optimal excitation of faint endogenous signals. However, obtaining these transform-limited short pulses remains a challenge, and previously proposed devices did not achieve an optimal pulse delivery. We present a study of fibered endomicroscope architecture with an efficient femtosecond pulse delivery and a high excitation level at the output of commercially available double-clad fibers (DCFs). The endomicroscope incorporates a module based on a grism line to compensate for linear and nonlinear effects inside the system. Simulations and experimental results are presented and compared to the literature. Experimentally, we obtained short pulses down to 24 fs at the fiber output, what represents to the best of our knowledge the shortest pulse duration ever obtained at the output of a nonlinear endoscopic system without postcompression. The choice of the optimal DCF among four possible commercial components is discussed and evaluated in regard to multiphoton excitation and fluorescence emission.

Optimization and characterization of nonlinear excitation and collection through a gradient-index lens for high-resolution nonlinear endomicroscopy

We report a study of gradient index (GRIN) lenses as a miniaturized micro-objective for in vivo imaging in the context of the development of a nonlinear endomicroscope. A numerical study of the parameters influencing the lateral resolution, excitation, and collection efficiency, when GRIN lens is coupled with a double clad fiber (DCF), is exposed. Four commercial DCFs, previously identified from the literature as potential endoscopic fibers, are simulated. Then, an experimental study characterizes two GRIN lenses (one commercial, one homemade) by their dispersion and nonlinear effects, potential intrinsic fluorescence, and use for fluorescence lifetime measurements. Images of neural cells from brain tissues of mice through a GRIN lens are presented.

Rhodamine B as an optical thermometer in cells focally exposed to infrared laser light or nanosecond pulsed electric fields.

The temperature-dependent fluorescence property of Rhodamine B was used to measure changes in temperature at the cellular level induced by either infrared laser light exposure or high intensity, ultrashort pulsed electric fields. The thermal impact of these stimuli were demonstrated at the cellular level in time and contrasted with the change in temperature observed in the extracellular bath. The method takes advantage of the temperature sensitivity of the fluorescent dye Rhodamine B which has a quantum yield linearly dependent on temperature. The thermal effects of different temporal pulse applications of infrared laser light exposure and of nanosecond pulsed electric fields were investigated. The temperature increase due to the application of nanosecond pulsed electric fields was demonstrated at the cellular level.

Characterization of fiber ultrashort pulse delivery for nonlinear endomicroscopy

In this work, we present a detailed characterization of a small-core double-clad photonic crystal fiber, dedicated and approved for in vivo nonlinear imaging endomicroscopy. A numerical and experimental study has been performed to characterize the excitation and collection efficiencies through a 5 m-long optical fiber, including the pulse duration and spectral shape. This was first done without any distal optics, and then the performances of the system were studied by using two kinds of GRIN lenses at the fiber output. These results are compared to published data using commercial double clad fibers and GRIN lenses.

Infrared neural stimulation induces intracellular Ca2+ release mediated by phospholipase C

The influence of infrared laser pulses on intracellular Ca2+ signaling was investigated in neural cell lines with fluorescent live cell imaging. The probe Fluo-4 was used to measure Ca2+ in HT22 mouse hippocampal neurons and nonelectrically excitable U87 human glioblastoma cells exposed to 50 to 500 ms infrared pulses at 1470 nm. Fluorescence recordings of Fluo-4 demonstrated that infrared stimulation induced an instantaneous intracellular Ca2+ transient with similar dose-response characteristics in hippocampal neurons and glioblastoma cells (half-maximal effective energy density EC50 of around 58 J.cm-2 ). For both type of cells, the source of the infrared-induced Ca2+ transients was found to originate from intracellular stores and to be mediated by phospholipase C and IP3 -induced Ca2+ release from the endoplasmic reticulum. The activation of phosphoinositide signaling by IR light is a new mechanism of interaction relevant to infrared neural stimulation that will also be widely applicable to nonexcitable cell types. The prospect of infrared optostimulation of the PLC/IP3 cell signaling cascade has many potential applications including the development of optoceutical therapeutics.

Dye-free determination of the focalization position for the hollow fiber flow field flow fractionation (HF5) of proteins

Proteins are separated in field flow fractionation (FFF) according to a well-established mechanism described as the “Normal or Brownian” mode. In the case of the sub-technique using a hollow fiber, the focalization/relaxation position can be observed visually only with a transparent holder and using dyes as samples. Whatever the choice of instrumentation, a dye-free method is proposed to determine the center of the zone from experimental fractograms by means of only two sample elutions. It is also possible to determine and model the kinematics of the sample toward the equilibrium focalization/relaxation position as well as the real dimensions of the fiber during the separation process.

Label free multiphoton imaging of human pulmonary tissues through two-meter-long microstructured fiber and multicore image-guide

This work deals with label free multiphoton imaging of the human lung tissue extra-cellular matrix (ECM) through optical fibers. Two devices were developed, the first one using distal scanning associated to a double clad large mode area (LMA) air-silica microstructured fiber, the second one using proximal scanning of a miniature multicore image guide (30000 cores inside a 0.8 mm diameter). In both cases, the main issue has been efficient linear and nonlinear distortion pre-compensation of excitation pulses. By inserting before the delivery fiber a compact (10 cm × 10 cm footprint) grisms-based stretcher (a grating in close contact with a prism) made of readily available commercial components, we achieved as short as 35-femtosecond-duration pulses that were temporally compressed at the direct exit of a 2-meter-long fiber. Interestingly, this femtosecond pulse fiber delivery device is also wavelength tunable over more than 100 nm inside the Ti: Sapphire emission band. With the help of distal scan system, those unique features allowed us to record elastin (through two-photon fluorescence) and collagen (through second harmonic generation) fibered network images. These images were obtained ex-vivo with only 15 mW @ 80 MHz of IR radiation delivered to the alveoli or bronchus tissues. 3D imaging with 400-μm-penetration depth inside the tissue was possible working with a 2-meter-long LMA fiber. With the help of proximal scanning, the miniature image guide allowed us to perform endoscopic real time microimaging of the ECM ex vivo.