A quantitative understanding of tissue morphogenesis requires description of the movements of individual cells in space and over time. In transparent embryos, such as C. elegans, fluorescently labeled nuclei can be imaged in three-dimensional time-lapse (4D) movies and automatically tracked through early cleavage divisions up to ~350 nuclei. A similar analysis of later stages of C. elegans development has been challenging owing to the increased error rates of automated tracking of large numbers of densely packed nuclei. We present Nucleitracker4D, a freely available software solution for tracking nuclei in complex embryos that integrates automated tracking of nuclei in local searches with manual curation. Using these methods, we have been able to track >99% of all nuclei generated in the C. elegans embryo. Our analysis reveals that ventral enclosure of the epidermis is accompanied by complex coordinated migration of the neuronal substrate. We can efficiently track large numbers of migrating nuclei in 4D movies of zebrafish cardiac morphogenesis, suggesting that this approach is generally useful in situations in which the number, packing or dynamics of nuclei present challenges for automated tracking.

To our knowledge, our solution is the first murmur classifier that focuses singularly on Still's murmur. Following further refinement and testing, the presented algorithm could reduce the number of children with Still's murmur referred unnecessarily to pediatric cardiologists.

Wnt signals orient mitotic spindles in development, but it remains unclear how Wnt signaling is spatially controlled to achieve precise spindle orientation. Here, we show that C. elegans syndecan (SDN-1) is required for precise orientation of a mitotic spindle in response to a Wnt cue. We find that SDN-1 is the predominant heparan sulfate (HS) proteoglycan in the early C. elegans embryo, and that loss of HS biosynthesis or of the SDN-1 core protein results in misorientation of the spindle of the ABar blastomere. The ABar and EMS spindles both reorient in response to Wnt signals, but only ABar spindle reorientation is dependent on a new cell contact and on HS and SDN-1. SDN-1 transiently accumulates on the ABar surface as it contacts C, and is required for local concentration of Dishevelled (MIG-5) in the ABar cortex adjacent to C. These findings establish a new role for syndecan in Wnt-dependent spindle orientation.

Abstract. The purpose of this work was to develop a clinically viable laparoscopic augmented reality (AR) system employing stereoscopic (3-D) vision, laparoscopic ultrasound (LUS), and electromagnetic (EM) tracking to achieve image registration. We investigated clinically feasible solutions to mount the EM sensors on the 3-D laparoscope and the LUS probe. This led to a solution of integrating an externally attached EM sensor near the imaging tip of the LUS probe, only slightly increasing the overall diameter of the probe. Likewise, a solution for mounting an EM sensor on the handle of the 3-D laparoscope was proposed. The spatial image-to-video registration accuracy of the AR system was measured to be 2.59 AE 0.58 mm and 2.43 AE 0.48 mm for the leftand right-eye channels, respectively. The AR system contributed 58-ms latency to stereoscopic visualization. We further performed an animal experiment to demonstrate the use of the system as a visualization approach for laparoscopic procedures. In conclusion, we have developed an integrated, compact, and EM tracking-based stereoscopic AR visualization system, which has the potential for clinical use. The system has been demonstrated to achieve clinically acceptable accuracy and latency. This work is a critical step toward clinical translation of AR visualization for laparoscopic procedures.

Purpose Common camera calibration methods employed in current laparoscopic augmented reality systems require the acquisition of multiple images of an entire checkerboard pattern from various poses. This lengthy procedure prevents performing laparoscope calibration in the operating room (OR). The purpose of this work was to develop a fast calibration method for electromagnetically (EM) tracked laparoscopes, such that calibration can be performed in the OR on demand. Methods We designed a mechanical tracking mount to uniquely and snugly position an EM sensor to an appropriate location on a conventional laparoscope. A tool named fCalib was developed to calibrate intrinsic camera parameters, distortion coefficients, and extrinsic parameters (transformation between the scope lens coordinate system and the EM sensor coordinate system) using a single image that shows an arbitrary portion of a special target pattern. For quick evaluation of calibration result in the OR, we integrated a tube phantom with fCalib and overlaid a virtual representation of the tube on the live video scene. Results We compared spatial target registration error between the common OpenCV method and the fCalib method in a laboratory setting. In addition, we compared the calibration re-projection error between the EM tracking-based fCalib and the optical tracking-based fCalib in a clinical setting. Our results suggested that the proposed method is comparable to the OpenCV method. However, changing the environment, e.g., inserting or removing surgical tools, would affect re-projection accuracy for the EM tracking-based approach. Computational time of the fCalib method averaged 14.0 s (range 3.5 s – 22.7 s). Conclusions We developed and validated a prototype for fast calibration and evaluation of EM tracked conventional (forward viewing) laparoscopes. The calibration method achieved acceptable accuracy and was relatively fast and easy to be performed in the OR on demand.