Monika Ritsch-Marte

Spiral phase contrast imaging in microscopy

We demonstrate an optical method for edge contrast enhancement in light microscopy. The method is based on holographic Fourier plane filtering of the microscopic image with a spiral phase element (also called vortex phase or helical phase filter) displayed as an off-axis hologram at a computer controlled high resolution spatial light modulator (SLM) in the optical imaging pathway. The phase hologram imprints a helical phase term of the form exp(i phi) on the diffracted light field in its Fourier plane. In the image plane, this results in a strong and isotropic edge contrast enhancement for both amplitude and phase objects.

Quantitative imaging of complex samples by spiral phase contrast microscopy

Recently a spatial spiral phase filter in a Fourier plane of a microscopic imaging setup has been demonstrated to produce edge enhancement and relief-like shadow formation of amplitude and phase samples. Here we demonstrate that a sequence of at least 3 spatially filtered images, which are recorded with different rotational orientations of the spiral phase plate, can be used to obtain a quantitative reconstruction of both, amplitude and phase information of a complex microscopic sample, i.e. an object consisting of mixed absorptive and refractive components. The method is demonstrated using a calibrated phase sample, and an epithelial cheek cell.

Cortical contractility triggers a stochastic switch to fast amoeboid cell motility.

Summary 3D amoeboid cell migration is central to many developmental and disease-related processes such as cancer metastasis. Here, we identify a unique prototypic amoeboid cell migration mode in early zebrafish embryos, termed stable-bleb migration. Stable-bleb cells display an invariant polarized balloon-like shape with exceptional migration speed and persistence. Progenitor cells can be reversibly transformed into stable-bleb cells irrespective of their primary fate and motile characteristics by increasing myosin II activity through biochemical or mechanical stimuli. Using a combination of theory and experiments, we show that, in stable-bleb cells, cortical contractility fluctuations trigger a stochastic switch into amoeboid motility, and a positive feedback between cortical flows and gradients in contractility maintains stable-bleb cell polarization. We further show that rearward cortical flows drive stable-bleb cell migration in various adhesive and non-adhesive environments, unraveling a highly versatile amoeboid migration phenotype.

Self-organized array of regularly spaced microbeads in a fiber-optical trap

The behavior of several simultaneously trapped, micrometer-sized particles in a fiber-optical trap consisting of two opposing single-mode fibers delivering counterpropagating, near-IR laser beams strongly depends on the size of the particles. Whereas beads that are considerably larger than the laser wavelength are pressed against each other in an axial line, smaller beads spontaneously arrange themselves into regular chains of equidistantly separated particles suspended in space with increasing separation for increasing bead diameter. A simple model based on self-organization by means of diffraction from the particles is capable of explaining the basic features of our experimental observations in the investigated range of bead diameters and refractive indices.

Wavefront correction of spatial light modulators using an optical vortex image

We present a fast and flexible non-interferometric method for the correction of small surface deviations on spatial light modulators, based on the Gerchberg-Saxton algorithm. The surface distortion information is extracted from the shape of a single optical vortex, which is created by the light modulator. The method can be implemented in optical tweezers systems for an optimization of trapping fields, or in an imaging system for an optimization of the point-spread-function of the entire image path.

Violation of a Bell inequality in two-dimensional orbital angular momentum state-spaces

We observe entanglement between photons in controlled super-position states of orbital angular momentum (OAM). By drawing a direct analogy between OAM and polarization states of light, we demonstrate the entangled nature of high order OAM states generated by spontaneous downconversion through violation of a suitable Clauser Horne Shimony Holt (CHSH)-Bell inequality. We demonstrate this violation in a number of two-dimensional subspaces of the higher dimensional OAM Hilbert space.

Near-perfect hologram reconstruction with a spatial light modulator

We present an implementation method for noiseless holographic projection of precalculated light fields with a spatial light modulator. In the reconstructed image, both the spatial amplitude and phase distributions can be programmed independently. This is achieved by diffracting the light from two successive phase holograms that are located in conjugate Fourier planes. The light path is folded such that the two corresponding phase masks can be displayed side by side at a single phase-only spatial light modulator. Such a device has relevant applications in holographic display-or projection systems, and for optical micromanipulation in laser tweezers.

mTHPC-mediated Photodynamic Diagnosis of Malignant Brain Tumors¶

Abstract Radical tumor resection is the basis for the prolonged survival of patients suffering from malignant brain tumors such as glioblastoma multiforme. We have carried out a phase-II study involving 22 patients with malignant brain tumors to assess the feasibility and the effectiveness of the combination of intraoperative photodynamic diagnosis and fluorescence-guided resection (FGR) mediated by the second-generation photosensitizer meta-tetrahydroxyphenylchlorin (mTHPC). In addition, intraoperative photodynamic therapy (PDT) was performed. Several commercially available fluorescence diagnostic systems were investigated for their applicability in clinical practice. We have adapted and optimized a diagnostic system that includes a surgical microscope, an excitation light source (filtered to 370–440 nm), a video camera detection system and a spectrometer for clear identification of the mTHPC fluorescence emission at 652 nm. Especially in regions of faint fluorescence, it turned out to be essential to maximize the spectral information by optimizing and matching the spectral properties of all components, such as excitation source, camera and color filters. To sum up, on the basis of 138 tissue samples derived from 22 tumor specimens, we have been able to achieve a sensitivity of 87.9% and a specificity of 95.7%. This study demonstrates that mTHPC-mediated intraoperative FGR followed by PDT is a highly promising concept in improving the radicality of tumor resection combined with a therapeutic approach.

Acoustic force spectroscopy

Force spectroscopy has become an indispensable tool to unravel the structural and mechanochemical properties of biomolecules. Here we extend the force spectroscopy toolbox with an acoustic manipulation device that can exert forces from subpiconewtons to hundreds of piconewtons on thousands of biomolecules in parallel, with submillisecond response time and inherent stability. This method can be readily integrated in lab-on-a-chip devices, allowing for cost-effective and massively parallel applications.

Wide-field coherent anti-Stokes Raman scattering microscopy

We suggest a variant of coherent anti-Stokes Raman scattering (CARS) microscopy, which is a nonlinear microscopy method that has recently proven very successful in functional imaging of living cells with vibrational contrast. Whereas existing CARS microscopy techniques involve confocal scanning, in our setup excitation and detection are performed in a wide-field (nonconfocal) geometry, similar to a combination of dark-field and epifluorescence microscopy. The use of a nanosecond laser system rather than the typically used pico- or femtosecond systems furthermore implies a high spectroscopic resolution for various Raman-active substances. We demonstrate the feasibility of the wide-field approach on test samples and discuss its prospects to become a complemental method to confocal scanning CARS microscopy.