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

3D laser nano-printing on fibre paves the way for super-focusing of multimode laser radiation

Multimode high-power laser diodes suffer from inefficient beam focusing, leading to a focal spot 10–100 times greater than the diffraction limit. This inevitably restricts their wider use in ‘direct-diode’ applications in materials processing and biomedical photonics. We report here a ‘super-focusing’ characteristic for laser diodes, where the exploitation of self-interference of modes enables a significant reduction of the focal spot size. This is achieved by employing a conical microlens fabricated on the tip of a multimode optical fibre using 3D laser nano-printing (also known as multi-photon lithography). When refracted by the conical surface, the modes of the fibre-coupled laser beam self-interfere and form an elongated narrow focus, usually referred to as a ‘needle’ beam. The multiphoton lithography technique allows the realisation of almost any optical element on a fibre tip, thus providing the most suitable interface for free-space applications of multimode fibre-delivered laser beams. In addition, we demonstrate the optical trapping of microscopic objects with a super-focused multimode laser diode beam thus rising new opportunities within the applications sector where lab-on-chip configurations can be exploited. Most importantly, the demonstrated super-focusing approach opens up new avenues for the ‘direct-diode’ applications in material processing and 3D printing, where both high power and tight focusing is required

Controlling waves in space and time for imaging and focusing in complex media

In complex media such as white paint and biological tissue, light encounters nanoscale refractive-index inhomogeneities that cause multiple scattering. Such scattering is usually seen as an impediment to focusing and imaging. However, scientists have recently used strongly scattering materials to focus, shape and compress waves by controlling the many degrees of freedom in the incident waves. This was first demonstrated in the acoustic and microwave domains using time reversal, and is now being performed in the optical realm using spatial light modulators to address the many thousands of spatial degrees of freedom of light. This approach is being used to investigate phenomena such as optical super-resolution and the time reversal of light, thus opening many new avenues for imaging and focusing in turbid medi

Generation of multiple Bessel beams for a biophotonics workstation

We present a simple method using an axicon and spatial light modulator to create multiple parallel Bessel beams and precisely control their individual positions in three dimensions. This technique is tested as an alternative to classical holographic beam shaping commonly used now in optical tweezers. Various applications of precise control of multiple Bessel beams are demonstrated within a single microscope giving rise to new methods for three-dimensional positional control of trapped particles or active sorting of micro-objects as well as "focus- free" photoporation of living cells. Overall this concept is termed a 'biophotonics workstation' where users may readily trap, sort and porate material using Bessel light modes in a microscope. (C) 2008 Optical Society of America.</p

Propagation characteristics of Airy beams:dependence upon spatial coherence and wavelength

We generate a broadband "white light" Airy beam and characterize the dependence of the beam properties on wavelength. Experimental results are presented showing that the beam's deflection coefficient and its characteristic length are wavelength dependent. In contrast the aperture coefficient is not wavelength dependent. However, this coefficient depends on the spatial coherence of the beam. We model this behaviour theoretically by extending the Gaussian-Schell model to describe the effect of spatial coherence on the propagation of Airy beams. The experimental results are compared to the model and good agreement is observed. (C) 2009 Optical Society of America</p

Large-field-of-view imaging by multi-pupil adaptive optics

Adaptive optics can correct for optical aberrations. We developed multi-pupil adaptive optics (MPAO), which enables simultaneous wavefront correction over a field of view of 450 ?? 450 ??m2 and expands the correction area to nine times that of conventional methods. MPAO&apos;s ability to perform spatially independent wavefront control further enables 3D nonplanar imaging. We applied MPAO to in vivo structural and functional imaging in the mouse brain.ope