Light sheet adaptive optics microscope for 3D live imaging

We report on the incorporation of adaptive optics (AO) into the imaging arm of a selective plane illumination microscope (SPIM). SPIM has recently emerged as an important tool for life science research due to its ability to deliver high-speed, optically sectioned, time-lapse microscope images from deep within in vivo selected samples. SPIM provides a very interesting system for the incorporation of AO as the illumination and imaging paths are decoupled and AO may be useful in both paths. In this paper, we will report the use of AO applied to the imaging path of a SPIM, demonstrating significant improvement in image quality of a live GFP-labeled transgenic zebrafish embryo heart using a modal, wavefront sensorless approach and a heart synchronization method. These experimental results are linked to a computational model showing that significant aberrations are produced by the tube holding the sample in addition to the aberration from the biological sample itself

Diamond machining of a single shot ellipsoidal focusing plasma mirror.

Plasma mirrors have become an important tool in high power laser physics due to their ability to suppress laser pre-pulses and amplified spontaneous emission allowing a cleaner and sharper rising edge pulse to be focused onto a target. A PMMA ellipsoidal plasma mirror used to increase the peak intensity of a high power laser pulses before it reaches the target is presented. The ellipse has been designed to increase by a factor 3, between input and output, the F-number of the beam, inducing in theory a factor 9 gain in peak intensity. Diamond machining, which is a technique capable of producing sub-micron accuracy on steep, freeform surfaces, is an ideal process for manufacturing these types of mirrors. In this paper, we discuss the diamond machining requirements to manufacture such near diffraction limited high numerical aperture mirrors

Comparison of closed loop and sensorless adaptive optics in widefield optical microscopy

We report on a closed loop widefield adaptive optics, optical microscopy system in which the feedback signal is provided by backscattered light from the sample acting as a guide star. The improvement in imaging performance is compared to an adaptive optics system controlled via an image optimisation routine commonly described as sensorless adaptive optics. The samples viewed were imaged without fluorescence to ensure that photobleaching and other potential variations did not affect the comparisons in system performance though the method is equally applicable for fluorescence microscopy. The closed loop system is self-optimising for different areas of the sample, using a common reference wavefront, with the accuracy of the loop being limited by variation across the sub-aperture images induced by guide star elongation. Optimisation using an image sharpness metric gives slightly sharper images but takes significantly longer. We thus believe that both wavefront sensor based closed loop AO and metric based optimisation have a role to play in AO for microscopy and that the method of backscattered light as a guide star has a great potential in the application of AO, particularly to optical coherence tomography

Towards freeform curved blazed gratings using diamond machining.

Concave blazed gratings greatly simplify the architecture of spectrographs by reducing the number of optical components. The production of these gratings using diamond-machining offers practically no limits in the design of the grating substrate shape, with the possibility of making large sag freeform surfaces unlike the alternative and traditional method of holography and ion etching. In this paper, we report on the technological challenges and progress in the making of these curved blazed gratings using an ultra-high precision 5 axes Moore-Nanotech machine. We describe their implementation in an integral field unit prototype called IGIS (Integrated Grating Imaging Spectrograph) where freeform curved gratings are used as pupil mirrors. The goal is to develop the technologies for the production of the next generation of low-cost, compact, high performance integral field unit spectrometers

Lightweighting design optimisation for additively manufactured mirrors

Design for additive manufacture (AM; 3D printing) is significantly different than design for subtractive machining. Although there are some limitations on the designs that can be printed, the increase in the AM design-space removes some of the existing challenges faced by the traditional lightweight mirror designs; for example, sandwich mirrors are just as easy to fabricate as open-back mirrors via AM, and they provide an improvement in structural rigidity. However, the ability to print a sandwich mirror as a single component does come with extra considerations; such as orientation upon the build plate and access to remove any temporary support material. This paper describes the iterations in optimisation applied to the lightweighting of a small, 84 mm diameter by 20 mm height, spherical concave mirror intended for CubeSat applications. The initial design, which was fabricated, is discussed in terms of the internal lightweighting design and the design constraints that were imposed by printing and post-processing. Iterations on the initial design are presented; these include the use of topology optimisation to minimise the total internal strain energy during mirror polishing and the use of lattices combined with thickness variation i.e. having a thicker lattice in strategic support locations. To assess the suitability of each design, finite element analysis is presented to quantify the print-through of the lightweighting upon the optical surface for a given mass reduction

Additively manufactured mirrors for CubeSats

Additive manufacturing (AM; 3D printing) is a fabrication process that builds an object layer-upon-layer and promotes the use of structures that would not be possible via subtractive machining. Prototype AM metal mirrors are increasingly being studied in order to exploit the advantage of the broad AM design-space to develop intricate lightweight structures that are more optimised for function than traditional open-back mirror lightweighting. This paper describes a UK Space Agency funded project to design and manufacture a series of lightweighted AM mirrors to fit within a 3 U CubeSat chassis. Five AM mirrors of identical design will be presented: two in aluminium (AlSi10Mg), two in nickel phosphorous (NiP) coated AlSi10Mg, and one in titanium (Ti64). For each material mirror pair, one is hand-polished (including the Ti64) and the other is diamond turned. Metrology data, surface form error and surface roughness, will be presented to compare and contrast the different materials and post-processing methods. To assess the presence of porosity, a frequent concern for AM materials, X-ray computed tomography measurements will be presented to highlight the location and density of pores within the mirror substrate; methods to mitigate the distribution of pores near the optical surface will be described. As a metric for success, the AlSi10Mg + NiP and AlSi10Mg mirrors should be suitable in terms of metrology data for visible and infrared applications respectively

Multimode fibre:Light-sheet microscopy at the tip of a needle

We also thank the UK Engineering and Physics Sciences Research Council for funding under grant EP/J01771X/1. Finally, we would like to thank EXCELLENT TEAMS (CZ.1.07/2.3.00/30.0005) from European Social Fund and CEITEC - Central European Institute of Technology (CZ.1.05/1.1.00/02.0068) from European Regional Development Fund for support.Light-sheet fluorescence microscopy has emerged as a powerful platform for 3-D volumetric imaging in the life sciences. Here, we introduce an important step towards its use deep inside biological tissue. Our new technique, based on digital holography, enables delivery of the light-sheet through a multimode optical fibre - an optical element with extremely small footprint, yet permitting complex control of light transport processes within. We show that this approach supports some of the most advanced methods in light-sheet microscopy: by taking advantage of the cylindrical symmetry of the fibre, we facilitate the wavefront engineering methods for generation of both Bessel and structured Bessel beam plane illumination. Finally, we assess the quality of imaging on a sample of fluorescent beads fixed in agarose gel and we conclude with a proof-of-principle imaging of a biological sample, namely the regenerating operculum prongs of Spirobranchus lamarcki.Publisher PDFPeer reviewe