Diffraction phase microscopy with white light

We present white light diffraction phase microscopy (wDPM) as a quantitative phase imaging method that combines the single shot measurement benefit associated with off-axis methods, high temporal phase stability associated with common path geometries, and high spatial phase sensitivity due to the white light illumination. We propose a spatiotemporal filtering method that pushes the limit of the pathlength sensitivity to the subangstrom level at practical spatial and temporal bandwidths. We illustrate the utility of wDPM with measurements on red blood cell morphology and HeLa cell growth over 18 hours.     

Fourier phase microscopy for investigation of biological structures and dynamics

By use of the Fourier decomposition of a low-coherence optical image field into two spatial components that can be controllably shifted in phase with respect to each other, a new high-transverse-resolution quantitative-phase microscope has been developed. The technique transforms a typical optical microscope into a quantitative-phase microscope, with high accuracy and a path-length sensitivity of lambda/5500, which is stable over several hours. The results obtained on epithelial and red blood cells demonstrate the potential of this instrument for quantitative investigation of the structure and dynamics associated with biological systems without sample preparation.     

Hilbert phase microscopy for investigating fast dynamics in transparent systems

We introduce Hilbert phase microscopy (HPM) as a novel optical technique for measuring high transverse resolution quantitative phase images associated with optically transparent objects. Because of its single-shot nature, HPM is suitable for investigating rapid phenomena that take place in transparent structures such as biological cells. The potential of this technique for studying biological systems is demonstrated with measurements of red blood cells, and its ability to quantify dynamic processes on a millisecond scale is exemplified with measurements of evaporating micrometer-sized water droplets.     

Modeling phase behavior for quantifying micro-pervaporation experiments

We present a theoretical model for the evolution of mixture concentrations in a micro-pervaporation device, similar to those recently presented experimentally. The described device makes use of the pervaporation of water through a thin PDMS membrane to build up a solute concentration profile inside a long microfluidic channel. We simplify the evolution of this profile in binary mixtures to a one-dimensional model which comprises two concentration-dependent coefficients. The model then provides a link between directly accessible experimental observations, such as the widths of dense phases or their growth velocity, and the underlying chemical potentials and phenomenological coefficients. It shall thus be useful for quantifying the thermodynamic and dynamic properties of dilute and dense binary mixtures.     

Imaging voltage-dependent cell motions with heterodyne Mach-Zehnder phase microscopy

We describe a heterodyne Mach-Zehnder interferometric microscope capable of quantitative phase imaging of biological samples with subnanometer sensitivity and frame rates up to 10 kHz. We use the microscope to image cultured neurons and measure nanometer-scale voltage-dependent motions in cells expressing the membrane motor protein prestin.     

Diffraction tomography without phase information

A modified form of diffraction tomography is presented in which measurements of the phase of the scattered field are replaced with measurements of the intensity on two planes beyond the scatterer. The new method is illustrated by an example.     

Atomic-scale structure of dislocations revealed by scanning tunneling microscopy and molecular dynamics

The intersection between dislocations and a Ag(111) surface has been studied using an interplay of scanning tunneling microscopy (STM) and molecular dynamics. Whereas the STM provides atomically resolved information about the surface structure and Burgers vectors of the dislocations, the simulations can be used to determine dislocation structure and orientation in the near-surface region. In a similar way, the subsurface structure of other extended defects can be studied. The simulations show dislocations to reorient the partials in the surface region leading to an increased splitting width at the surface, in agreement with the STM observations. Implications for surface-induced cross slip are discussed.     

Diffraction from phase modulated apertures

We apply the Rayleigh-Sommerfeld diffraction integral to solve the diffracted fields of periodically phase-modulated apertures. We show that the diffraction of small phase modulated apertures (with few refractive index modulation periods) behaves as for phase gratings, but the sharp diffraction peaks may already appear in the Fresnel region behind the aperture. Thus, these components may serve as lensless miniature interconnection gratings in micro-optics.     

Diffraction theory of third-harmonic large-aperture nonlinear microscopy

In the scalar approximation of the diffraction imaging theory, the main characteristics of a large-aperture nonlinear microscope operating at the third harmonic in transmitted or reflected light are determined. Interference of the obtained images with the third-harmonic background generated by the cover glass of a sample is studied. Problems of selection of the optimal version of microscopy are discussed. Original Russian Text ? G.M. Svishchev, 2008, published in Optika i Spektroskopiya, 2008, Vol. 104, No. 5, pp. 873–878.     

Diffraction regimes of phase gratings

The occurrence of Raman-Nath regime and Bragg regime diffraction by planar phase gratings is rigorously analyzed. It is shown that the published indicators for a Raman-Nath regime are unsatisfactory. An attempt to redefine the Raman-Nath region is given in this paper.     

Phase-shifting Zernike phase contrast microscopy for quantitative phase measurement

Zernike phase contrast microscopy is extended and combined with a phase-shifting mechanism to perform quantitative phase measurements of microscopic objects. Dozens of discrete point light sources on a ring are constructed for illumination. For each point light source, three different levels of point-like phase steps are designed, which are alternatively located along a ring on a silica plate to perform phase retardation on the undiffracted (dc) component of the object waves. These three levels of the phase steps are respectively selected by rotating the silica plate. Thus, quantitative evaluation of phase specimens can be performed via phase-shifting mechanism. The proposed method has low "halo" and "shade-off" effects, low coherent noise level, and high lateral resolution due to the improved illumination scheme.     

Compact,common path quantitative phase microscopic techniques for imaging cell dynamics

Microscopy using visible electromagnetic radiation can be used to investigate living cells in various environments. But bright field microscopy only provides two-dimensional (2D) intensity distribution at a single object plane. One of the ways to retrieve object height/thickness information is to employ quantitative phase microscopic (QPM) techniques. Interferometric QPM techniques are widely used for this. Digital holographic microscopy (DHM) is one of the state-of-the-art methods for quantitative three-dimensional (3D) imaging. Usually it is implemented in two-beam geometry, which is prone to mechanical vibrations. But to study dynamics of objects like red blood cells, one needs temporal stability much better than the fluctuations of the object, which the two-beam geometry fails to deliver. One way to overcome this hurdle is to use self-referencing techniques, in which a portion of the object beam will act as the reference beam. Here the development of self-referencing QPM techniques is described along with the results.     

Spectral-domain phase microscopy

Broadband interferometry is an attractive technique for the detection of cellular motions because it provides depth-resolved phase information via coherence gating. We present a phase-sensitive technique called spectral-domain phase microscopy (SDPM). SDPM is a functional extension of spectral-domain optical coherence tomography that allows for the detection of nanometer-scale motions in living cells. The sensitivity of the technique is demonstrated, and its calibration is verified. A shot-noise limit to the displacement sensitivity of this technique is derived. Measurement of cellular dynamics was performed on spontaneously beating cardiomyocytes isolated from chick embryos.     

Diffraction efficiency and electron microscopy of large amplitude holographic gratings

The diffraction efficiency of Ag coated holographic gratings was measured as a function of the grating amplitude, H, for a He-Ne laser incident at 30° and 70° in the plane perpendicular to the grating grooves. The grating amplitude was determined using a scanning electron microscope. It was found that the diffraction efficiency of s polarized light increased monotonically with increasing H for H < 250 nm. In contrast, for the same amplitude range, the diffraction efficiency of p polarized light oscillated through two maxima at H = 90 nmand 220 nm. These results were compared to the theoretical calculations of Heitmann and found to agree within 10% for H < 60 nm in the s polarization case and for H < 15 nm in the p polarization case. For larger H, the data deviated significantly, presumably due to a breakdown of the small amplitude assumption on which the calculations were based.     

Controllable tomography phase microscopy

Tomography phase microscopy (TPM) is a new microscopic method that can quantitatively yield the volumetric 3D distribution of a sample׳s refractive index (RI), which is significant for cell biology research. In this paper, a controllable TPM system is introduced. In this system a circulatory phase-shifting method and piezoelectric ceramic are used which enable the TPM system to record the 3D RI distribution at a more controllable speed, from 1 to 40 fps, than in the other TPM systems reported. The resolution of the RI distribution obtained by this controllable TPM is much better than that in images recorded by phase contrast microscopy and interference tomography microscopy. The realization of controllable TPM not only allows for the application of TPM to the measurement of kinds of RI sample, but also contributes to academic and technological support for the practical use of TPM.     

Quantitative optical phase microscopy

We present a new method for the extraction of quantitative phase data from microscopic phase samples by use of partially coherent illumination and an ordinary transmission microscope. The technique produces quantitative images of the phase profile of the sample without phase unwrapping. The technique is able to recover phase even in the presence of amplitude modulation, making it significantly more powerful than existing methods of phase microscopy. We demonstrate the technique by providing quantitatively correct phase images of well-characterized test samples and show that the results obtained for more-complex samples correlate with structures observed with Nomarski differential interference contrast techniques.     

Full-field swept-source phase microscopy

We present a full-field phase microscopy technique for quantitative nanoscale surface profiling of samples in reflection. This technique utilizes swept-source optical coherence tomography in a full-field common path interferometer for phase-stable cross-sectional acquisition without scanning. Subwavelength variations in surface sample features are measured without interference from spurious reflections by processing the interferometric phase at a selected depth plane, providing a 1.3 nm stability for high signal-to-noise ratio surface features. Nanoscale imaging was demonstrated by measuring the location of receptor sites on a DNA assay biochip and the surface topography of erythrocytes in a blood smear.     

Diffraction anomalies in moving periodic structure

Special method is presented for treatment of moving periodic structure. The analyzing of the simplest model reveals the speed-dependent diffraction anomalies. This physical effect only occurs under the slow-wave driving and when transverse resonance condition satisfied. The theory and the results may have significance in interdisciplinary fields associated with moving periodic structure.