Holographic mensuration of suspended particles in aquatic systems

The distribution and dynamics of aggregates in the aquatic environment play an important role in the modelling of biogeochemical processes. Previous work on aggregates in the ocean (e.g. sedimentary 'marine snow' particles), which vary in size from tens of microns to several millimetres, has used electronic counting or conventional photography coupled with image analysis. Here we describe a non-destructive in situ approach by use of holographic mensuration, hologrammetry, that affords greater scope and higher accuracy for the enumeration, sizing, and spatial distribution determination of aggregate particles. By means of two complimentary techniques, in-line and offaxis transmission holography, we present the initial experiments conducted in our laboratory and discuss the preliminaiy results from real image analysis

Off-axis transmission holographic system for recording aquatic particles

We describe a holographic system for recording particles suspended in water. The hologram plate is located in air, separated from the test tank by an air/glass/water boundary. The holographic emulsion is therefore unaffected by adverse aquatic conditions within the tank (i.e. surface contamination, non-uniform swelling). The design geometry is intended to minimise the aberrations that arise from recording subjects located in water and replaying their hologram image in air. Third order aberrations, most crucially spherical aberration and astigmatism, are suppressed to give an experimental resolution of 7 lp/mm using USAF 1951 target in water 600mm from the boundary. Particles (plankton species) in the sub-millimeter to several millimeters size range are observed at planar sections within the recording volume by visual inspection of the hologram replayed in real image mode

In situ off-axis holography of marine plankton

We present an off-axis transmission holographic technique for recording marine plankton in situ within a test tank of 36,000 ml with a pulsed laser in a 40 ns interval. The holographic plate is located in air and is therefore unaffected by aquatic conditions that may cause emulsion degradation (e.g. non-uniform swelling and surface contamination). The reference beam traverses a path in air only, and thus remains unaffected by dense concentrations of plankton. Third order aberrations, notably spherical aberration and astigmatism, are suppressed to yield an experimental resolution of 7 lp/mm (70 micrometres) with a USAF 1951 target located 600 mm in water from the observation window. Plankton particle counts examined by real image reconstruction show a strong correlation with duplicate samples examined under a microscope

A data extraction system for underwater particle holography

Pulsed laser holography is an extremely powerful technique for the study of particle fields as it allows instantaneous, noninvasive high-resolution recording of substantial volumes. By replaying the real image one can obtain the size, shape, position and - if multiple exposures are made - velocity of every object in the recorded field. Manual analysis of large volumes containing thousands of particles is, however, an enormous and time-consuming task, with operator fatigue an unpredictable source of errors. Clearly the value of holographic measurements also depends crucially on the quality of the reconstructed image: not only will poor resolution degrade size and shape measurements, but aberrations such as coma and astigmatism can change the perceived centroid of a particle, affecting position and velocity measurements. For large-scale applications of particle field holography, specifically the in situ recording of marine plankton with 'HoloCam,' we have developed an automated data extraction system that can be readily switched between the in-line and off-axis geometries and provides optimised reconstruction from holograms recorded underwater. As a videocamera is automatically stepped through the 200 by 200 by 1000mm sample volume, image processing and object tracking routines locate and extract particle images for further classification by a separate software module

Subwavelength hyperspectral THz studies of articular cartilage

This is the final version of the article. Available from the publisher via the DOI in this record.Terahertz-spectroscopy probes dynamics and spectral response of collective vibrational modes in condensed phase, which can yield insight into composition and topology. However, due to the long wavelengths employed (λ = 300 μm at 1THz), diffraction limited imaging is typically restricted to spatial resolutions around a millimeter. Here, we demonstrate a new form of subwavelength hyperspectral, polarization-resolved THz imaging which employs an optical pattern projected onto a 6 μm-thin silicon wafer to achieve near-field modulation of a co-incident THz pulse. By placing near-field scatterers, one can measure the interaction of object with the evanescent THz fields. Further, by measuring the temporal evolution of the THz field a sample's permittivity can be extracted with 65 μm spatial resolution due to the presence of evanescent fields. Here, we present the first application of this new approach to articular cartilage. We show that the THz permittivity in this material varies progressively from the superficial zone to the deep layer, and that this correlates with a change in orientation of the collagen fibrils that compose the extracellular matrix (ECM) of the tissue. Our approach enables direct interrogation of the sample's biophysical properties, in this case concerning the structure and permittivity of collagen fibrils and their anisotropic organisation in connective tissue.The research presented in this work was funded by QinetiQ & EPSRC under iCase award 12440575 and grant number EP/K041215/1

The development and sea trials of a subsea holographic camera for large volume in-situ recording of marine organisms

We describe the development, construction and sea testing of an underwater holographic camera (HoloCam) for in situ recording of marine organisms and particles in large volumes of sea water. HoloCam comprises a laser, power supply, holographic recording optics and plate holders, a water-tight housing and a support frame. Added to this are control electronics such that the entire camera is remotely operable and controllable from ship or dock-side. Uniquely the camera can simultaneously record both in-line and off-axis holograms using a pulsed frequency doubled Nd-YAG laser. In-line holography is capable of producing images of organisms with a resolution of better than 10 Pm (at concentrations up to a few thousand per cubic centimetre at the smallest sizes). Off-axis holograms of aquatic systems of up to 50,000 cm3 volume, have been recorded. Following initial laboratory testing, the holo-camera was evaluated in an observation tank and ultimately was tested in Loch Etive, Scotland. In-line and off-axis holograms were recorded to a depth of 100 m. We will present results on the test dives and evaluation of the camera performance

HoloCam: A subsea holographic camera for recording marine organisms and particles

The HoloCam system is a major component of a multi-national multi-discipline project known as HoloMar (funded by the European Commission under the MAST III initiative). The project is concerned with the development of pulsed laser holography to analyse and monitor the populations of living organisms and inanimate particles within the world's oceans. We describe here the development, construction and evaluation of a prototype underwater camera, the purpose of which is to record marine organisms and particles, in-situ. Recording using holography provides several advantages over conventional sampling methods in that it allows non-intrusive, non-destructive, high-resolution imaging of large volumes (up to 10^5 cm^3) in three dimensions. The camera incorporates both in-line and off-axis holographic techniques, which allows particles from a few micrometres to tens of centimetres to be captured. In tandem with development of the HoloCam, a dedicated holographic replay system and an automated data extraction and image processing facility are being developed. These will allow, optimisation of the images recorded by the camera, identification of species and particle concentration plotting

A holographic system for subsea recording and analysis of plankton and other marine particles

We report here details of the design, development, initial testing and field-deployment of the HOLOMAR system for in-situ subsea holography and analysis of marine plankton and nonliving particles. HOLOMAR comprises a submersible holographic camera ("HoloCam") able to record in-line and off-axis holograms at depths down to 100 m, together with specialised reconstruction hardware ("HoloScan") linked to custom image processing and classification software. The HoloCam consists of a laser and power supply, holographic recording optics and holographic plate holders, a water-tight housing and a support frame. It utilises two basic holographic geometries, in-line and off-axis such that a wide range of species, sizes and concentrations can be recorded. After holograms have been recorded and processed they are reconstructed in full three-dimensional detail in air in a dedicated replay facility. A computer-controlled microscope, using video cameras to record the image at a given depth, is used to digitise the scene. Specially written software extracts a binarised image of an object in its true focal plane and is classified using a neural network. The HoloCam was deployed on two separate cruises in a Scottish sea loch (Loch Etive) to a depth of 100 m and over 300 holograms were recorded

High-resolution in situ holographic recording and analysis of marine organisms and particles (HOLOMAR)

We report on the development of a fully- unctioning, prototype, underwater holographic camera (holo-camera) for holographic recording of large-volumes of sea water containing marine plankton and seston within the upper water column The overriding benefit of holographic imaging over other measurement techniques is that it allows non-intrusive and non-destructive, in-situ, recording of living organisms and inanimate particles in their natural environment. Because of the inherently high resolution of holography, its threedimensional imaging properties and the ability to perform "optical sectioning" on the image, it allows identification of particular organisms together with the extraction of sue and relative positional information This information, in turn, affords the ability to gain knowledge of the behaviour of marine biological communities, their relationship with each other and with the particles with which they interact