Scattering in biological tissues is a major barrier for in vivo optical imaging of all but the most superficial structures. Progress toward overcoming the distortions caused by scattering in turbid ...media has been made by shaping the excitation wavefront to redirect power into a single point in the imaging plane. However, fast, non-invasive determination of the required wavefront compensation remains challenging. Here, we introduce a quickly converging algorithm for non-invasive scattering compensation, termed DASH, in which holographic phase stepping interferometry enables new phase information to be updated after each measurement. This leads to rapid improvement of the wavefront correction, forming a focus after just one measurement iteration and achieving an order of magnitude higher signal enhancement at this stage than the previous state-of-the-art. Using DASH, we demonstrate two-photon fluorescence imaging of microglia cells in highly turbid mouse hippocampal tissue down to a depth of 530 μm.
The spectral dispersion of ultrashort pulses allows the simultaneous focusing of light in both space and time, which creates so-called spatiotemporal foci. Such space-time coupling may be combined ...with the existing holographic techniques to give a further dimension of control when generating focal light fields. In the present study, 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. By exploiting the pulse front tilt generated at focus when applying simultaneous spatial and temporal focusing (SSTF), it is possible to overlap neighboring 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 the non-linear laser fabrication of glass.
Single molecule localization microscopy (SMLM) has the potential to resolve structural details of biological samples at the nanometer length scale. Compared to room temperature experiments, SMLM ...performed under cryogenic temperature achieves higher photon yields and, hence, higher localization precision. However, to fully exploit the resolution it is crucial to account for the anisotropic emission characteristics of fluorescence dipole emitters with fixed orientation. In case of slight residual defocus, localization estimates may well be biased by tens of nanometers. We show here that astigmatic imaging in combination with information about the dipole orientation allows to extract the position of the dipole emitters without localization bias and down to a precision of 1 nm, thereby reaching the corresponding Cramér Rao bound. The approach is showcased with simulated data for various dipole orientations, and parameter settings realistic for real life experiments.
We present a method to increase the number of simultaneously imaged focal planes in diffractive multi-plane imaging. We exploit the chromatic properties of diffraction by using multicolor LED ...illumination and demonstrate time-synchronous imaging of up to 21 focal planes.We discuss the possibilities and limits given by the use of a liquid crystal spatial light modulator to display the diffractive patterns. The method is suitable for wide-field transmission and reflection microscopy.
Femtosecond laser fabrication of controlled three dimensional structures deep in the bulk of diamond is facilitated by a dual adaptive optics system. A deformable mirror is used in parallel with a ...liquid crystal spatial light modulator to compensate the extreme aberrations caused by the refractive index mismatch between the diamond and the objective immersion medium. It is shown that aberration compensation is essential for the generation of controlled micron-scale features at depths greater than 200 μm, and the dual adaptive optics approach demonstrates increased fabrication efficiency relative to experiments using a single adaptive element.
We demonstrate that a parallel aligned liquid crystal on silicon (PA-LCOS) spatial light modulator (SLM) without any attached color mask can be used as a full color display with white light ...illumination. The method is based on the wavelength dependence of the (voltage controlled) birefringence of the liquid crystal pixels. Modern SLMs offer a wide range over which the birefringence can be modulated, leading (in combination with a linear polarizer) to several intensity modulation periods of a reflected light wave as a function of the applied voltage. Because of dispersion, the oscillation period strongly depends on the wavelength. Thus each voltage applied to an SLM pixel corresponds to another reflected color spectrum. For SLMs with a sufficiently broad tuning range, one obtains a color palette (i.e., a "color lookup-table"), which allows one to display color images. An advantage over standard liquid crystal displays (LCDs), which use color masks in front of the individual pixels, is that the light efficiency and the display resolution are increased by a factor of three.
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 a new method for femtosecond laser writing of optical-fiber-compatible glass waveguides, namely spherical phase-induced multicore waveguide (SPIM-WG), which addresses this challenging task with three-dimensional on-chip light control. Fabricating in the heating regime with high scanning speed, precise deformation of cross-sections is still 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 fiber. SPIM-WG devices were easily fabricated that were able to perform on-chip beam rotation through varying angles, or manipulate the 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 nonuniform modes; examples include circular, elliptical modes, and asymmetric modes from ppKTP (periodically poled potassium titanyl phosphate) 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 fiber also paves the way toward packaged photonic integrated circuitry, which usually needs input and output fiber connections.
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