40 research outputs found
Three-dimensional imaging of direct-written photonic structures
Third harmonic generation microscopy has been used to analyze the morphology
of photonic structures created using the femtosecond laser direct-write
technique. Three dimensional waveguide arrays and waveguide-Bragg gratings
written in fused-silica and doped phosphate glass were investigated. A
sensorless adaptive optical system was used to correct the optical aberrations
occurring in the sample and microscope system, which had a lateral resolution
of less than 500 nm. This non-destructive testing method creates volume
reconstructions of photonic devices and reveals details invisible to other
linear microscopy and index profilometry techniques.Comment: 8 pages, 3 color figures, 2 hyper-linked animation
Four-dimensional light shaping: manipulating ultrafast spatio-temporal foci in space and time
Spectral dispersion of ultrashort pulses allows simultaneous focusing of
light in both space and time creating so-called spatio-temporal foci. Such
space-time coupling may be combined with existing holographic techniques to
give a further dimension of control when generating focal light fields. It is
shown that a phase-only hologram placed in the pupil plane of an objective and
illuminated by a spatially chirped ultrashort pulse can be used to generate
three dimensional arrays of spatio-temporally focused spots. Exploiting the
pulse front tilt generated at focus when applying simultaneous spatial and
temporal focusing (SSTF), it is possible to overlap neighbouring foci in time
to create a smooth intensity distribution. The resulting light field displays a
high level of axial confinement, with experimental demonstrations given through
two-photon microscopy and non-linear laser fabrication of glass
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Deconvolution approach for 3D scanning microscopy with helical phase engineering.
RESCH (refocusing after scanning using helical phase engineering) microscopy is a scanning technique using engineered point spread functions which provides volumetric information. We present a strategy for processing the collected raw data with a multi-view maximum likelihood deconvolution algorithm, which inherently comprises the resolution gain of pixel-reassignment microscopy. The method, which we term MD-RESCH (for multi-view deconvolved RESCH), achieves in our current implementation a 20% resolution advantage along all three axes compared to RESCH and confocal microscopy. Along the axial direction, the resolution is comparable to that of image scanning microscopy. However, because the method inherently reconstructs a volume from a single 2D scan, a significantly higher optical sectioning becomes directly visible to the user, which would otherwise require collecting multiple 2D scans taken at a series of axial positions. Further, we introduce the use of a single-helical detection PSF to obtain an increased post-acquisition refocusing range. We present data from numerical simulations as well as experiments to confirm the validity of our approach
Characterisation of the dynamic behaviour of lipid droplets in the early mouse embryo using adaptive harmonic generation microscopy
<p>Abstract</p> <p>Background</p> <p>Lipid droplets (LD) are organelles with an important role in normal metabolism and disease. The lipid content of embryos has a major impact on viability and development. LD in Drosophila embryos and cultured cell lines have been shown to move and fuse in a microtubule dependent manner. Due to limitations in current imaging technology, little is known about the behaviour of LD in the mammalian embryo. Harmonic generation microscopy (HGM) allows one to image LD without the use of exogenous labels. Adaptive optics can be used to correct aberrations that would otherwise degrade the quality and information content of images.</p> <p>Results</p> <p>We have built a harmonic generation microscope with adaptive optics to characterise early mouse embryogenesis. At fertilization, LD are small and uniformly distributed, but in the implanting blastocyst, LD are larger and enriched in the invading giant cells of the trophectoderm. Time-lapse studies reveal that LD move continuously and collide but do not fuse, instead forming aggregates that subsequently behave as single units. Using specific inhibitors, we show that the velocity and dynamic behaviour of LD is dependent not only on microtubules as in other systems, but also on microfilaments. We explore the limits within which HGM can be used to study living embryos without compromising viability and make the counterintuitive finding that 16 J of energy delivered continuously over a period of minutes can be less deleterious than an order of magnitude lower energy delivered dis-continuously over a period of hours.</p> <p>Conclusions</p> <p>LD in pre-implantation mouse embryos show a previously unappreciated complexity of behaviour that is dependent not only on microtubules, but also microfilaments. Unlike LD in other systems, LD in the mouse embryo do not fuse but form aggregates. This study establishes HGM with adaptive optics as a powerful tool for the study of LD biology and provides insights into the photo-toxic effects of imaging embryos.</p
Optofluidic adaptive optics in multi-photon microscopy
Adaptive optics in combination with multi-photon techniques is a powerful
approach to image deep into a specimen. Remarkably, virtually all adaptive
optics schemes today rely on wavefront modulators which are reflective,
diffractive, or both. This, however, can pose a severe limitation for
applications. Here, we present a fast and robust sensorless adaptive optics
scheme adapted for transmissive wavefront modulators. We study our scheme in
numerical simulations and in experiments with a novel, optofluidic wavefront
shaping device which is transmissive, refractive, polarisation-independent and
broadband. We demonstrate scatter correction of two-photon-excited fluorescence
images of microbeads as well as brain cells and benchmark our device against a
liquid-crystal spatial light modulator. Our method and technology could open
new routes for adaptive optics in scenarios where previously the restriction to
reflective and diffractive devices may have staggered innovation and progress
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Spectral image scanning microscopy
For decades, the confocal microscope has represented one of the dominant imaging systems in biomedical imaging at sub-cellular length scales. Recently, however, it has increasingly been replaced by a related, but more powerful successor technique termed image scanning microscopy (ISM). In this article, we present ISM capable of measuring spectroscopic information such as that contained in fluorescence or Raman images. Compared to established confocal spectroscopic imaging systems, our implementation offers similar spectral resolution, but higher spatial resolution and detection efficiency. Color sensitivity is achieved by a grating placed in the detection path in conjunction with a camera collecting both spatial and spectral information. The multidimensional data is processed using multi-view maximum likelihood image reconstruction. Our findings are supported by numerical simulations and experiments on micro beads and double-stained HeLa cells.</p
Position clamping in a holographic counterpropagating optical trap
Optical traps consisting of two counterpropagating, divergent beams of light allow relatively high forces to be exerted along the optical axis by turning off one beam, however the axial stiffness of the trap is generally low due to the lower numerical apertures typically used. Using a high speed spatial light modulator and CMOS camera, we demonstrate 3D servocontrol of a trapped particle, increasing the stiffness from 0.004 to 1.5μNm<sup>−1</sup>. This is achieved in the “macro-tweezers” geometry [Thalhammer, J. Opt. 13, 044024 (2011); Pitzek, Opt. Express 17, 19414 (2009)], which has a much larger field of view and working distance than single-beam tweezers due to its lower numerical aperture requirements. Using a 10×, 0.2NA objective, active feedback produces a trap with similar effective stiffness to a conventional single-beam gradient trap, of order 1μNm<sup>−1</sup> in 3D. Our control loop has a round-trip latency of 10ms, leading to a resonance at 20Hz. This is sufficient bandwidth to reduce the position fluctuations of a 10μm bead due to Brownian motion by two orders of magnitude. This approach can be trivially extended to multiple particles, and we show three simultaneously position-clamped beads
On-chip beam rotators, polarizers and adiabatic mode converters through low-loss waveguides with variable cross-sections
Photonics integrated circuitry would benefit considerably from the ability to arbitrarily control waveguide cross-sections with high precision and low loss, in order to provide more degrees of freedom in manipulating propagating light. Here, we report on a new optical-fibres-compatible glass waveguide by femtosecond laser writing, namely spherical phase induced multi-core waveguide (SPIM-WG), which addresses this challenging task with three dimensional on-chip light control. Precise deformation of cross-sections is achievable along the waveguide, with shapes and sizes finely controllable of high resolution in both horizontal and vertical transversal directions. We observed that these waveguides have high refractive index contrast of 0.017, low propagation loss of 0.14 dB/cm, and very low coupling loss of 0.19 dB coupled from a single mode fibre. SPIM-WG devices were easily fabricated that were able to perform on-chip beam rotation through varying angles, or manipulate polarization state of propagating light for target wavelengths. We also demonstrated SPIM-WG mode converters that provide arbitrary adiabatic mode conversion with high efficiency between symmetric and asymmetric non-uniform modes; examples include circular, elliptical modes and asymmetric modes from ppKTP waveguides which are generally applied in frequency conversion and quantum light sources. Created inside optical glass, these waveguides and devices have the capability to operate across ultra-broad bands from visible to infrared wavelengths. The compatibility with optical fibre also paves the way toward packaged photonic integrated circuitry, which usually needs input and output fibre connections