Cell shape dynamics: from waves to migration.

We observe and quantify wave-like characteristics of amoeboid migration. Using the amoeba Dictyostelium discoideum, a model system for the study of chemotaxis, we demonstrate that cell shape changes in a wave-like manner. Cells have regions of high boundary curvature that propagate from the leading edge toward the back, usually along alternating sides of the cell. Curvature waves are easily seen in cells that do not adhere to a surface, such as cells that are electrostatically repelled from surfaces or cells that extend over the edge of micro-fabricated cliffs. Without surface contact, curvature waves travel from the leading edge to the back of a cell at -35 µm/min. Non-adherent myosin II null cells do not exhibit these curvature waves. At the leading edge of adherent cells, curvature waves are associated with protrusive activity. Like regions of high curvature, protrusive activity travels along the boundary in a wave-like manner. Upon contact with a surface, the protrusions stop moving relative to the surface, and the boundary shape thus reflects the history of protrusive motion. The wave-like character of protrusions provides a plausible mechanism for the zig-zagging of pseudopods and for the ability of cells both to swim in viscous fluids and to navigate complex three dimensional topography.

A grating-based single-shot x-ray phase contrast and diffraction method for in vivo imaging.

PURPOSE: The purpose of this study is to develop a single-shot version of the grating-based phase contrast x-ray imaging method and demonstrate its capability of in vivo animal imaging. Here, the authors describe the principle and experimental results. They show the source of artifacts in the phase contrast signal and optimal designs that minimize them. They also discuss its current limitations and ways to overcome them. METHODS: A single lead grid was inserted midway between an x-ray tube and an x-ray camera in the planar radiography setting. The grid acted as a transmission grating and cast periodic dark fringes on the camera. The camera had sufficient spatial resolution to resolve the fringes. Refraction and diffraction in the imaged object manifested as position shifts and amplitude attenuation of the fringes, respectively. In order to quantify these changes precisely without imposing a fixed geometric relationship between the camera pixel array and the fringes, a spatial harmonic method in the Fourier domain was developed. The level of the differential phase (refraction) contrast as a function of hardware specifications and device geometry was derived and used to guide the optimal placement of the grid and object. Both ex vivo and in vivo images of rodent extremities were collected to demonstrate the capability of the method. The exposure time using a 50 W tube was 28 s. RESULTS: Differential phase contrast images of glass beads acquired at various grid and object positions confirmed theoretical predictions of how phase contrast and extraneous artifacts vary with the device geometry. In anesthetized rats, a single exposure yielded artifact-free images of absorption, differential phase contrast, and diffraction. Differential phase contrast was strongest at bone-soft tissue interfaces, while diffraction was strongest in bone. CONCLUSIONS: The spatial harmonic method allowed us to obtain absorption, differential phase contrast, and diffraction images, all from a single raw image and is feasible in live animals. Because the sensitivity of the method scales with the density of the gratings, custom microfabricated gratings should be superior to off-the-shelf lead grids.

Selective imaging of nano-particle contrast agents by a single-shot x-ray diffraction technique.

Iron oxide nano-particles have very different x-ray diffraction properties from tissue. They can be clearly visualized against suppressed tissue background in a single-shot x-ray diffraction imaging technique. This technique is able to acquire both diffraction and absorption images from a single grating-modulated projection image through analysis in the spatial frequency domain. We describe the use of two orthogonal transmission gratings to selectively retain diffraction signal from iron oxide particles that are larger than a threshold size, while eliminating the background signal from soft tissue and bone. This approach should help the tracking of functionalized particles in cell labeling and targeted therapy.

Simultaneous myocardial strain and dark-blood perfusion imaging using a displacement-encoded MRI pulse sequence.

The purpose of this study is to develop and evaluate a displacement-encoded pulse sequence for simultaneous perfusion and strain imaging. Displacement-encoded images in two to three myocardial slices were repeatedly acquired using a single-shot pulse sequence for 3 to 4 min, which covers a bolus infusion of Gadolinium contrast. The magnitudes of the images were T(1) weighted and provided quantitative measures of perfusion, while the phase maps yielded strain measurements. In an acute coronary occlusion swine protocol (n = 9), segmental perfusion measurements were validated against microsphere reference standard with a linear regression (slope 0.986, R(2) = 0.765, Bland-Altman standard deviation = 0.15 mL/min/g). In a group of ST-elevation myocardial infarction patients (n = 11), the scan success rate was 76%. Short-term contrast washout rate and perfusion are highly correlated (R(2) = 0.72), and the pixelwise relationship between circumferential strain and perfusion was better described with a sigmoidal Hill curve than linear functions. This study demonstrates the feasibility of measuring strain and perfusion from a single set of images.

Single-shot x-ray differential phase-contrast and diffraction imaging using two-dimensional transmission gratings.

We describe an x-ray differential phase-contrast imaging method based on two-dimensional transmission gratings that are directly resolved by an x-ray camera. X-ray refraction and diffraction in the sample lead to variations of the positions and amplitudes of the grating fringes on the camera. These effects can be quantified through spatial harmonic analysis. The use of 2D gratings allows differential phase contrast in several directions to be obtained from a single image. When compared to previous grating-based interferometry methods, this approach obviates the need for multiple exposures and separate measurements for different directions and thereby accelerates imaging speed.