Colin Sheppard
Colin Sheppard obtained his PhD degree in Engineering from University of Cambridge (1973). He developed an ultra-high vacuum scanning electron diffraction system, and applied it to observation of the oxidation of metal surfaces, under the supervision of Professor Sir Charles Oatley, who constructed the first scanning electron microscope. He spent two years in industry, at English Electric Valve Company, researching on negative electron affinity silicon photocathodes. He then went to Oxford, where he worked with Professor Rudi Kompfner, inventor of the traveling wave tube and former director of Bell Labs at Holmdel. He was University Lecturer in Engineering Science at the University of Oxford, and Official Fellow and Tutor in Engineering at Pembroke College. He was awarded the DSc degree in Physical Sciences from University of Oxford in 1986. In 1989, he moved to Australia, where he was Professor of Physics and Head of the Physical Optics Department at the University of Sydney. From 2003, he was Professor and Head of the Department of Bioengineering at the National University of Singapore, where he also held joint appointments in Diagnostic Radiology, Biological Sciences and the NUS Graduate School (NGS). From 2012 to 2017 he was at the Italian Institute of Technology in Genoa as Senior Scientist.
Professor Sheppard is currently Honorary Professorial Fellow at the University of Wollongong, Australia; External Collaborator and Visiting Scientist with the Italian Institute of Technology, Genoa, Italy; and Supernumerary Fellow of Pembroke College Oxford.
He has held visiting professorships at many different institutions, including UC-Berkeley, MIT, EPFL, TU-Delft, Warsaw University of Technology, University of Erlangen-Nürnberg, University of Jena, Autonomous University of Barcelona , University of Buenos Aires, University of the Philippines, University of Western Australia, and University of Queensland, as well as fellowships at Stanford University, Tokyo University, UNSW, University of Besançon, and Institut Fresnel Marseille.
He was elected Fellow of OSA, SPIE, IEE, Institute of Physics, Japan Society for Applied Physics, and the Royal Microscopical Society. He served as Vice-President of the International Commission for Optics (ICO), and as President of the International Society for Optics Within Life Sciences (OWLS). He was awarded the Italian Society for Optics and Photonics Galileo Galilei Medal, Institute of Physics Optics and Photonics Division Prize, Max Planck Bonhöffer Medal, Humboldt Research Award, ISATA Mercedes Award, National Physical Laboratory Metrology Award, Prince of Wales Award for Industrial Innovation, British Technology Group Academic Enterprise Award, and the IEE Gyr and Landis Prize.
He developed an early laser microscope (1975), patented scanning microscopy using Bessel beams (1977), published the first demonstration of scanning two-photon microscopy (by SHG) (1977), published the first paper on Bessel-Gauss beams (1978), proposed two-photon fluorescence and CARS microscopy (1978), launched the first commercial confocal microscope (1982), developed the first confocal microscope with computer control and storage (1983), proposed scanning microscopy using a detector array with pixel reassignment, now known as image scanning microscopy (1988). He has made substantial contributions in the fields of diffraction theory, beam propagation, pulse propagation, scattering and inverse scattering, 3D image formation, and polarization algebra.
Professor Sheppard is currently Honorary Professorial Fellow at the University of Wollongong, Australia; External Collaborator and Visiting Scientist with the Italian Institute of Technology, Genoa, Italy; and Supernumerary Fellow of Pembroke College Oxford.
He has held visiting professorships at many different institutions, including UC-Berkeley, MIT, EPFL, TU-Delft, Warsaw University of Technology, University of Erlangen-Nürnberg, University of Jena, Autonomous University of Barcelona , University of Buenos Aires, University of the Philippines, University of Western Australia, and University of Queensland, as well as fellowships at Stanford University, Tokyo University, UNSW, University of Besançon, and Institut Fresnel Marseille.
He was elected Fellow of OSA, SPIE, IEE, Institute of Physics, Japan Society for Applied Physics, and the Royal Microscopical Society. He served as Vice-President of the International Commission for Optics (ICO), and as President of the International Society for Optics Within Life Sciences (OWLS). He was awarded the Italian Society for Optics and Photonics Galileo Galilei Medal, Institute of Physics Optics and Photonics Division Prize, Max Planck Bonhöffer Medal, Humboldt Research Award, ISATA Mercedes Award, National Physical Laboratory Metrology Award, Prince of Wales Award for Industrial Innovation, British Technology Group Academic Enterprise Award, and the IEE Gyr and Landis Prize.
He developed an early laser microscope (1975), patented scanning microscopy using Bessel beams (1977), published the first demonstration of scanning two-photon microscopy (by SHG) (1977), published the first paper on Bessel-Gauss beams (1978), proposed two-photon fluorescence and CARS microscopy (1978), launched the first commercial confocal microscope (1982), developed the first confocal microscope with computer control and storage (1983), proposed scanning microscopy using a detector array with pixel reassignment, now known as image scanning microscopy (1988). He has made substantial contributions in the fields of diffraction theory, beam propagation, pulse propagation, scattering and inverse scattering, 3D image formation, and polarization algebra.
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Books by Colin Sheppard
Over the last decade, several excellent books and reviews on confocal microscopy have been published, but there has been a noticeable gap in the availability of a small handbook that introduces the interested student or research worker to this important microscopical technique, and that illustrates how it might benefit their own research. It is this gap that we hope this Handbook, the latest in the Royal Microscopical Society Microscopy Handbooks series, will fill.
Starting from first principles, this Handbook explains to the reader what a confocal microscope is, how it is constructed and used what its benefits. are, and. why i.ts imaging performance is superior to'that of a conventional optical mIcroscope. It discusses multiparameter confocal fluorescence microscopy, describes digital image processing and animation of 3D confocal images, illustrates applications of confocal
microscopy in both the biomedical and the materials sciences and concludes with a discussion of future developments in this new area of microscopy. Richly illustrated with colour micrographs and diagrams chosen for their clarity and didactic quality, this book also contains an up-to-date bibliography of the most informative publications on confocal
microscopy, a catalogue of World Wide Web sites of relevance, and a listing of the names and addresses of confocal microscope and fluorescence filter manufacturers, image processing software vendors, and reagent suppliers. We hope that you will find it useful.
Book Chapters by Colin Sheppard
has been investigated theoretically. Imaging can be described by a three-dimensional (3-D) coherent transfer function (CTF), which has contributions from the different wavelength components. From this, two dimensional imaging can be described using the 2-D CTF obtained as a projection of the 3-D CTF. For very low numerical aperture axial imaging results from the limited coherence length of the light source. Interference microscopy results in an optical sectioning property similar to that in confocal microscopy. Thus for intermediate values of numerical aperture, axial imaging is a combination of coherence gating and confocal sectioning, for which a paraxial theory can be used. At very high numerical apertures it is necessary to use a full high aperture theory. These theoretical treatments can be used to model images of known structures, and to estimate expected imaging performance.
Over the last decade, several excellent books and reviews on confocal microscopy have been published, but there has been a noticeable gap in the availability of a small handbook that introduces the interested student or research worker to this important microscopical technique, and that illustrates how it might benefit their own research. It is this gap that we hope this Handbook, the latest in the Royal Microscopical Society Microscopy Handbooks series, will fill.
Starting from first principles, this Handbook explains to the reader what a confocal microscope is, how it is constructed and used what its benefits. are, and. why i.ts imaging performance is superior to'that of a conventional optical mIcroscope. It discusses multiparameter confocal fluorescence microscopy, describes digital image processing and animation of 3D confocal images, illustrates applications of confocal
microscopy in both the biomedical and the materials sciences and concludes with a discussion of future developments in this new area of microscopy. Richly illustrated with colour micrographs and diagrams chosen for their clarity and didactic quality, this book also contains an up-to-date bibliography of the most informative publications on confocal
microscopy, a catalogue of World Wide Web sites of relevance, and a listing of the names and addresses of confocal microscope and fluorescence filter manufacturers, image processing software vendors, and reagent suppliers. We hope that you will find it useful.
has been investigated theoretically. Imaging can be described by a three-dimensional (3-D) coherent transfer function (CTF), which has contributions from the different wavelength components. From this, two dimensional imaging can be described using the 2-D CTF obtained as a projection of the 3-D CTF. For very low numerical aperture axial imaging results from the limited coherence length of the light source. Interference microscopy results in an optical sectioning property similar to that in confocal microscopy. Thus for intermediate values of numerical aperture, axial imaging is a combination of coherence gating and confocal sectioning, for which a paraxial theory can be used. At very high numerical apertures it is necessary to use a full high aperture theory. These theoretical treatments can be used to model images of known structures, and to estimate expected imaging performance.
regard to their behaviour at high apertures.
of the transfer function.
all having Gaussian pupil function is developed. This assumption is useful as the expressions may be evaluated analytically. It is shown that Type 2 microscopes exhibit superior performance to those of Type 1. Effects of defocus are considered. It is found that defocus can be used in a Type 2 microscope to observe phase information without the limitation in resolution associated with stopping down the collector of a conventional microscope. It is also found that a Type 2 microscope discriminates against light scattered by parts of the object outside of the focal plane, allowing observation of detail within a thick object.
If the effective source and effective detector are coherent and incoherent, respectively, the microscope (Type 1) is of the form of the scanning transmission electron microscope (STEM) . If the effective source and the effective detector are both coherent, the microscope (Type 2) is of the form of the scanning acoustic microscope. Scanning optical microscopes of both these types may be con- structed. The effect of using a Type 2 scanning microscope in dark-field is discussed. This arrangement has the advantage over the Type 1 dark-field microscope that imaging for weak contrast specimens may be made linear .
In the axoneme, dynein induced sliding of apposing microtubules is responsible
for the motility of cilia and flagella. Although it has been speculated that the
interaction between dyne in and microtubules in the axoneme is spatiotemporally
correlated with the bend propagation, such correlation has never been
experimentally demonstrated. A key challenge in studying the correlation is to
detect structural changes in the beating axoneme with high enough spatial and
temporal resolution.
In this study, we demonstrate that a high-aperture dark-field microscope can
detect minute structural changes that occur along the length of a beating
axoneme. We show slow-motion recordings of beating axonemes and
quantitative image analysis combined with theoretical mode ling of the dark field
imaging mode to extract structural information gleaned from changes in
scattering power of axoneme segments. Based on dark-field imaging of
demembranated beating axonemes of sea urchin spermatozoa, we found
changes in the scattered intensity that propagated with the flagellar wave.
Interestingly, we detected changes in scattering power of fragmented axonemes
that were exposed to different nucleotide conditions using caged ATP
photolysis. These results can be explained assuming changes in the effective
diameter of the axoneme. Our combined results lead us to propose regional
structural differences along beating axonemes triggered by ATP hydrolysis.