15 research outputs found
Multi-photon attenuation-compensated light-sheet fluorescence microscopy
We thank the UK Engineering and Physical Sciences Research Council for funding (grants EP/P030017/1 and EP/R004854/1), the European Union’s Horizon 2020 Framework Programme (H2020) (675512, BE-OPTICAL), the Danish Council for Independent Research (DFF FTP grant 7017-00021), and the Otto Mønsted Foundation (grant 19-70-0109).Attenuation of optical fields owing to scattering and absorption limits the penetration depth for imaging. Whilst aberration correction may be used, this is difficult to implement over a large field-of-view in heterogeneous tissue. Attenuation-compensation allows tailoring of the maximum lobe of a propagation-invariant light field and promises an increase in depth penetration for imaging. Here we show this promising approach may be implemented in multi-photon (two-photon) light-sheet fluorescence microscopy and, furthermore, can be achieved in a facile manner utilizing a graded neutral density filter, circumventing the need for complex beam shaping apparatus. A “gold standard” system utilizing a spatial light modulator for beam shaping is used to benchmark our implementation. The approach will open up enhanced depth penetration in light-sheet imaging to a wide range of end users.Publisher PDFPeer reviewe
Realization of laterally nondispersing ultrabroadband Airy pulses
We present the measurements of the spatiotemporal impulse response of a system creating nondispersing Airy pulses, i.e., ultrabroadband Airy beams whose main lobe size remains constant over propagation. A custom refractive element with a continuous surface profile was used to impose the cubic phase on the input beam. The impulse response of the Airy pulse generator was spatiotemporally characterized by applying a white-light spatial-spectral interferometry setup based on the SEA TADPOLE technique. The results were compared with the theoretical model and previously spatiotemporally characterized Airy pulses generated by a spatial light modulator. (C) 2014 Optical Society of Americ
Separation of the rotational contribution in fluorescence correlation experiments
The theory of fluorescence correlation spectroscopy is reexamined with the aim of separating the contribution of rotational diffusion. Under constant excitation, fluorescence correlation experiments are characterized by three polarizations: one of the incident beam and two of the two photon detectors. A set of experiments of different polarizations is proposed for study. From the results of the experiments the isotropic factor of the fluorescence intensity correlation functions can be determined, which is independent of the rotational motion of the sample molecule. This function can be used to represent each fluorescence intensity correlation function as the product of the isotropic and the rotational factors. The theory is illustrated by an experiment in which rotational diffusion of porcine pancreatic lipase labeled with Texas Red was observed Texas Red is a label that allows precise fluorescence correlation experiments even in the nanosecond time range
Measuring the suppression of ultrashort pulses into Airy-Bessel light bullets with almost single-cycle temporal resolution
We have achieved ∼3 fs and ∼3 μm resolutions for full spatio-temporal characterization of impulse responses of optical systems using a white-light spectral interferometry set-up and demonstrate how a circular diffraction grating temporally focuses an Airy-Bessel wave-packet