Conjunctival UV autofluorescence - prevalence and risk factors

Purpose: Autofluorescence of ultraviolet (UV) light has been shown to occur in localised areas of the bulbar conjunctiva, which map to active cellular changes due to UV and environmental exposure. This study examined the presence of conjunctival UV autofluorescence in eye care practitioners (ECPs) across Europe and the Middle East and its associated risk factors. Method: Images were captured of 307 ECPs right eyes in the Czech Republic, Germany, Greece, Kuwait, Netherlands, Sweden, Switzerland, United Arab Emirates and the United Kingdom using a Nikon D100 camera and dual flash units through UV filters. UV autofluorescence was outlined using ImageJ software and the nasal and temporal area quantified. Subjects were required to complete a questionnaire on their demographics and lifestyle including general exposure to UV and refractive correction. Results: Average age of the subjects was 38.5±12.2 years (range 19-68) and 39.7% were male. Sixty-two percent of eyes had some conjunctival damage as indicated by UV autofluorescence. The average area of damage was higher (p=0.005) nasally (2.95±4.52mm2) than temporally (2.19±4.17mm2). The area of UV damage was not related to age (r=0.03, p=0.674), gender (p=0.194), self-reported sun exposure lifestyle (p>0.05), geographical location (p=0174), sunglasses use (p>0.05) or UV-blocking contact lens use (p>0.05), although it was higher in those wearing contact lenses with minimal UV-blocking and no spectacles (p=0.015). The area of UV damage was also less nasally in those who wore contact lenses and spectacles compared to those with no refractive correction use (p=0.011 nasal; p=0.958 temporal). Conclusion: UV conjunctival damage is common even in Europe, Kuwait and UAE, and among ECPs. The area of damage appears to be linked with the use of refractive correction, with greater damage nasally than temporally which may be explained by the peripheral light focusing effect.

Ophthalmic engineering:the development of novel instrumentation to further research in the field

The principle theme of this thesis is the advancement and expansion of ophthalmic research via the collaboration between professional Engineers and professional Optometrists. The aim has been to develop new and novel approaches and solutions to contemporary problems in the field. The work is sub divided into three areas of investigation; 1) High technology systems, 2) Modification of current systems to increase functionality, and 3) Development of smaller more portable and cost effective systems. High Technology Systems: A novel high speed Optical Coherence Tomography (OCT) system with integrated simultaneous high speed photography was developed achieving better operational speed than is currently available commercially. The mechanical design of the system featured a novel 8 axis alignment system. A full set of capture, analysis, and post processing software was developed providing custom analysis systems for ophthalmic OCT imaging, expanding the current capabilities of the technology. A large clinical trial was undertaken to test the dynamics of contact lens edge interaction with the cornea in-vivo. The interaction between lens edge design, lens base curvature, post insertion times and edge positions was investigated. A novel method for correction of optical distortion when assessing lens indentation was also demonstrated. Modification of Current Systems: A commercial autorefractor, the WAM-5500, was modified with the addition of extra hardware and a custom software and firmware solution to produce a system that was capable of measuring dynamic accommodative response to various stimuli in real time. A novel software package to control the data capture process was developed allowing real time monitoring of data by the practitioner, adding considerable functionality of the instrument further to the standard system. The device was used to assess the accommodative response differences between subjects who had worn UV blocking contact lens for 5 years, verses a control group that had not worn UV blocking lenses. While the standard static measurement of accommodation showed no differences between the two groups, it was determined that the UV blocking group did show better accommodative rise and fall times (faster), thus demonstrating the benefits of the modification of this commercially available instrumentation. Portable and Cost effective Systems: A new instrument was developed to expand the capability of the now defunct Keeler Tearscope. A device was developed that provided a similar capability in allowing observation of the reflected mires from the tear film surface, but with the added advantage of being able to record the observations. The device was tested comparatively with the tearscope and other tear film break-up techniques, demonstrating its potential. In Conclusion: This work has successfully demonstrated the advantages of interdisciplinary research between engineering and ophthalmic research has provided new and novel instrumented solutions as well as having added to the sum of scientific understanding in the ophthalmic field.

Immunochemical detection of UV-induced DNA damage and repair

The application of an antiserum to ultraviolet radiation (UVR)-damaged DNA is presented. A novel experimental system was employed to ascertain the limits of detection for this antiserum. Using a DNA standard containing a known amount of dimer, the limits of detection were found to be 0.9 fmol of dimer. This was compared to a limit of 20-50 fmol dimer using gas chromatography-mass spectrometry (GC-MS). Induction of thymine dimers in DNA following UVR exposure, as assessed using this antiserum in an enzyme-linked immunosorbent assay (ELISA), was compared with GC-MS measurements. The ELISA method successfully demonstrated the induction of lesions in DNA irradiated either with UVC or UVB, although despite high sensitivity, no discernible binding was seen to UVA-irradiated DNA. The antiserum was also shown to be applicable to immunocytochemistry, localising damage in the nuclei of UVR exposed keratinocytes in culture. The ability of the antiserum to detect DNA damage in skin biopsies of individuals exposed to sub-erythemal doses of UVR was also demonstrated. Moreover, the subsequent removal of this damage, as evidenced by a reduction in antiserum staining, was noted in sections of biopsies taken in the hours following irradiation. © 2003 Elsevier B.V. All rights reserved.

Immunochemical detection of sequence-specific modifications to DNA induced by UV light

Sequence specificity of antibodies to UV-damaged DNA has not been described previously. The antisera investigated here were specific for UV-modified DNA and were absolutely dependent upon the presence of thymine residues. Using a series of oligonucleotides in competition ELISA, increased inhibition was observed with increasing chain length of UV-polythymidylate. A minimum of three adjacent thymines was required for effective inhibition; alone, dimers of thymine were poor antigens. Although UV-irradiated poly(dC) was not antigenic, cytosines could partially replace thymines within the smallest effective epitope (T-T-T) with a high degree of sequence specificity, not previously described. The main epitope induced by UV was formed from adjacent thymines and either a 3' or a 5' pyrimidine.

UV femtosecond laser inscribes a 300 nm period nanostructure in a pure fused silica

We report on the first recording of a periodic structure of ∼150 nm pitch in a permanently moving sample of a pure fused silica using the tightly focused, 82 nJ, 267 nm, 300 fs, 1 kHz laser pulses. © 2007 IOP Publishing Ltd.

Advanced UV inscribed fibre grating structures and applications in optical sensing and laser systems

This thesis presents detailed investigation of UV inscribed fibre grating based devices and novel developments in the applications of such devices in optical sensing and fibre laser systems. The major contribution of this PhD programme includes the systematic study on fabrication, spectral characteristics and applications of different types of UV written in-fibre gratings such as Type I and IA Fibre Bragg Gratings (FBGs), Chirped Fibre Bragg Gratings (CFBGs) and Tilted Fibre Gratings (TFGs) with small, large and 45º tilted structures inscribed in normal silica fibre. Three fabrication techniques including holographic, phase-mask and blank beam exposure scanning, which were employed to fabricate a range of gratings in standard single mode fibre, are fully discussed. The thesis reports the creation of smart structures with self-sensing capability by embedding FBG-array sensors in Al matrix composite. In another part of this study, we have demonstrated the particular significant improvements made in sensitising standard FBGs to the chemical surrounding medium by inducing microstructure to the grating by femtosecond (fs) patterning assisted chemical etching technique. Also, a major work is presented for the investigation on the structures, inscription methods and spectral Polarisation Dependent Loss (PDL) and thermal characteristics of different angle TFGs. Finally, a very novel application in realising stable single polarisation and multiwavelength switchable Erbium Doped Fibre Lasers (EDFLs) using intracavity polarisation selective filters based on TFG devices with tilted structures at small, large and exact 45° angles forms another important contribution of this thesis.

Writing and characterisation methods for UV-photosensitivity based devices and their applications

The underlying work to this thesis focused on the exploitation and investigation of photosensitivity mechanisms in optical fibres and planar waveguides for the fabrication of advanced integrated optical devices for telecoms and sensing applications. One major scope is the improvement of grating fabrication specifications by introducing new writing techniques and the use of advanced characterisation methods for grating testing. For the first time the polarisation control method for advanced grating fabrication has successfully been converted to apodised planar waveguide fabrication and the development of a holographic method for the inscription of chirped gratings at arbitrary wavelength is presented. The latter resulted in the fabrication of gratings for pulse-width suppression and wavelength selection in diode lasers. In co-operation with research partners a number of samples were tested using optical frequency domain and optical low coherence reflectometry for a better insight into the limitations of grating writing techniques. Using a variety of different fabrication methods, custom apodised and chirped fibre Bragg gratings were written for the use as filter elements for multiplexer-demultiplexer devices, as well as for short pulse generation and wavelength selection in telecommunication transmission systems. Long period grating based devices in standard, speciality and tapered fibres are presented, showing great potential for multi-parameter sensing. One particular scope is the development of vectorial curvature and refractive index sensors with potential for medical, chemical and biological sensing. In addition the design of an optically tunable Mach-Zehnder based multiwavelength filter is introduced. The discovery of a Type IA grating type through overexposure of hydrogen loaded standard and Boron-Germanium co-doped fibres strengthened the assumption of UV-photosensitivity being a highly non-linear process. Gratings of this type show a significantly lower thermal sensitivity compared to standard gratings, which makes them useful for sensing applications. An Oxford Lasers copper-vapour laser operating at 255 nm in pulsed mode was used for their inscription, in contrast to previous work using CW-Argon-Ion lasers and contributing to differences in the processes of the photorefractive index change

Water diffusion into UV inscripted long period grating in microstructured polymer fibre

A long period grating was photoinscribed step-by-step in microstructured poly(methyl methacrylate) fiber for the first time using a continuous wave HeCd laser at 325 nm, irradiating the fiber with a power of 1 mW. The grating had a length of 2 cm and a period of 1 mm. A series of cladding mode coupling resonances were observed throughout the spectral region studied of 600 to 1100 nm. The resonance wavelengths were shown to be sensitive to the diffusion of water into the fiber.

Point-by-point inscription of 250 nm period structure in bulk fused silica by tightly focused femtosecond UV pulses

By conducting point-by-point inscription in a continuously moving slab of pure fused silica at the optimal depth (170νm depth below the surface), we have fabricated a 250nm period nanostructure with 30nJ, 300fs, 1kHz pulses from a frequency-tripled Ti:sapphire laser. This is the smallest value for the inscribed period yet reported, and has been achieved with radical improvement in the quality of the inscribed nanostructures in comparison with previous reports.

UV femtosecond laser inscribes a 300 nm period nanostructure in a pure fused silica

We report on the first recording of a periodic structure of ~150 nm pitch in a permanently moving sample of a pure fused silica using the tightly focused, 82 nJ, 267 nm, 300 fs, 1 kHz laser pulses.