4D modelling of fluid mechanics in the zebrafish embryonic heart.

Abnormal blood flow mechanics can result in pathological heart malformation, underlining the importance of understanding embryonic cardiac fluid mechanics. In the current study, we performed image-based computational fluid dynamics simulation of the zebrafish embryonic heart ventricles and characterized flow mechanics, organ dynamics, and energy dynamics in detail. 4D scans of 5 days post-fertilization embryonic hearts with GFP-labelled myocardium were acquired using line-scan focal modulation microscopy. This revealed that the zebrafish hearts exhibited a wave-like contractile/relaxation motion from the inlet to the outlet during both systole and diastole, which we showed to be an energy efficient configuration. No impedance pumping effects of pressure and velocity waves were observed. Due to its tube-like configuration, inflow velocities were higher near the inlet and smaller at the outlet and vice versa for outflow velocities. This resulted in an interesting spatial wall shear stress (WSS) pattern where WSS waveforms near the inlet and those near the outlet were out of phase. There was large spatial variability in WSS magnitudes. Peak WSS was in the range of 47.5-130 dyne/cm2 at the inflow and outflow tracts, but were much smaller, in the range of 4-11 dyne/cm2, in the mid-ventricular segment. Due to very low Reynolds number and the highly viscous environment, intraventricular pressure gradients were high, suggesting substantial energy losses of flow through the heart.

Effects of pupil filter patterns in line-scan focal modulation microscopy.

Line-scan focal modulation microscopy (LSFMM) is an emerging imaging technique that affords high imaging speed and good optical sectioning at the same time. We present a systematic investigation into optimal design of the pupil filter for LSFMM in an attempt to achieve the best performance in terms of spatial resolutions, optical sectioning, and modulation depth. Scalar diffraction theory was used to compute light propagation and distribution in the system and theoretical predictions on system performance, which were then compared with experimental results.

Line-scan focal modulation microscopy.

We report the development of a line-scan focal modulation microscope (LSFMM) that is capable of high-speed image acquisition ( > 40 ?? fps ) with uncompromised optical sectioning capability. The improved background rejection and axial resolution of this imaging modality, enabled by focal modulation, are quantified with three-dimensional imaging data obtained from fluorescent beads. The signal-to-background ratio for the LSFMM system is one- to two-orders of magnitude higher than that for line-scanning confocal systems when imaging deep (up to 100 ?m) into a turbid medium of optical properties similar to biological tissues. The imaging performance of LSFMM, in terms of both spatial and temporal resolutions, is further demonstrated with in vivo imaging experiments with live zebrafish larvae.

Augmented line-scan focal modulation microscopy for multi-dimensional imaging of zebrafish heart in vivo.

Multi-dimensional fluorescence imaging of live animal models demands strong optical sectioning, high spatial resolution, fast image acquisition, and minimal photobleaching. While conventional laser scanning microscopes are capable of deep penetration and sub-cellular resolution, they are generally too slow and causing excessive photobleaching for volumetric or time-lapse imaging. We demonstrate the performance of an augmented line-scan focal modulation microscope (aLSFMM), a high-speed imaging platform that affords above video-rate imaging speed by the use of line scanning. Exceptional background rejection is accomplished by combining a confocal slit with focal modulation. The image quality is further improved by merging the information from simultaneously acquired focal modulation and confocal images. Such a hybrid imaging scheme makes it possible to use very low power excitation light in high-speed imaging, and therefore leads to reduced photobleaching that is desirable for three-dimensional (3D) and four-dimensional (4D) in vivo image acquisition.

The conductivity of neonatal piglet skulls.

We report the first measured values of conductivities for neonatal mammalian skull samples. We measured the average radial (normal to the skull surface) conductivity of fresh neonatal piglet skull samples at 1 kHz and found it to be around 30 mS m(-1) at ambient room temperatures of about 23 °C. Measurements were made on samples of either frontal or parietal cranial bone, using a saline-filled cell technique. The conductivity value we observed was approximately twice the values reported for adult skulls (Oostendorp et al 2000 IEEE Trans. Biomed. Eng. 47 1487-92) using a similar technique, but at a frequency of around 5 Hz. Further, we found that the conductivity of skull fragments increased linearly with thickness. We found evidence that this was related to differences in composition between the frontal and parietal bone samples tested, which we believe is because frontal bones contained a larger fraction of higher conductivity cancellous bone material.

Silica nanocapsules of fluorescent conjugated polymers and superparamagnetic nanocrystals for dual-mode cellular imaging.

We describe here a facile and benign synthetic strategy to integrate the fluorescent behavior of conjugated polymers and superparamagnetic properties of iron oxide nanocrystals into silica nanocapsules, forming a new type of bifunctional magnetic fluorescent silica nanocapsule (BMFSN). The resultant BMFSNs are uniform, colloidally stable in aqueous medium, and exhibit the desired dual functionality of fluorescence and superparamagnetism in a single entity. Four conjugated polymers with different emissions were used to demonstrate the versatility of employing this class of fluorescent materials for the preparation of BMFSNs. The applicability of BMFSNs in cellular imaging was studied by incubating them with human liver cancer cells, the result of which demonstrated that the cells could be visualized by dual-mode fluorescence and magnetic resonance imaging. Furthermore, the superparamagnetic behavior of the BMFSNs was exploited for in vitro magnetic-guided delivery of the nanocapsules into the cancer cells, thereby highlighting their potential for targeting biomedical applications.