In Vivo Visualization of Calcium Transients during Fertilization and Early Development in C. elegans.

Calcium is an important signaling molecule during the oocyte-to-embryo transition (OET) and early embryogenesis. The hermaphroditic nematode Caenorhabditis elegans provides several unique advantages for the study of the OET as it is transparent and has an ordered gonad that produces one mature oocyte every ~23 min at 20 °C. We have modified the genetically encoded calcium indicator jGCaMP7s to fluorescently indicate the moment of fertilization within a living organism. We have termed this reporter "CaFE" for Calcium during Fertilization in C. elegans. The CaFE reporter was engineered into a safe harbor locus in single copy, has no significant impact on physiology or fecundity, and produces a robust signal upon fertilization. Here, a series of protocols is presented for utilizing the CaFE reporter as an in vivo tool for dissecting the OET and embryogenesis. We include methods to synchronize worms, examine the effects of RNAi knockdown, mount worms for imaging, and to visualize calcium in oocytes and embryos. Additionally, we present the generation of additional worm strains to aid in this type of analysis. Demonstrating the utility of the CaFE reporter to visualize the timing of fertilization, we report that double ovulation occurs when ipp-5 is targeted by RNAi and that only the first oocyte undergoes immediate fertilization. Furthermore, the discovery of single-cell calcium transients during early embryogenesis is reported here, demonstrating that the CaFE reporter persists into early development. Importantly, the CaFE reporter in worms is simple enough to use for incorporation into course-based undergraduate research (CURE) laboratory classes. The CaFE reporter, coupled with the ordered gonad and ease of RNAi in worms, facilitates inquiry into the cell-cell dynamics required to regulate internal fertilization and early embryogenesis.

Integrated optical frequency division for microwave and mmWave generation.

The generation of ultra-low-noise microwave and mmWave in miniaturized, chip-based platforms can transform communication, radar and sensing systems1-3. Optical frequency division that leverages optical references and optical frequency combs has emerged as a powerful technique to generate microwaves with superior spectral purity than any other approaches4-7. Here we demonstrate a miniaturized optical frequency division system that can potentially transfer the approach to a complementary metal-oxide-semiconductor-compatible integrated photonic platform. Phase stability is provided by a large mode volume, planar-waveguide-based optical reference coil cavity8,9 and is divided down from optical to mmWave frequency by using soliton microcombs generated in a waveguide-coupled microresonator10-12. Besides achieving record-low phase noise for integrated photonic mmWave oscillators, these devices can be heterogeneously integrated with semiconductor lasers, amplifiers and photodiodes, holding the potential of large-volume, low-cost manufacturing for fundamental and mass-market applications13.

Towards high-power, high-coherence, integrated photonic mmWave platform with microcavity solitons.

Millimetre-wave (mmWave) technology continues to draw great interest due to its broad applications in wireless communications, radar, and spectroscopy. Compared to pure electronic solutions, photonic-based mmWave generation provides wide bandwidth, low power dissipation, and remoting through low-loss fibres. However, at high frequencies, two major challenges exist for the photonic system: the power roll-off of the photodiode, and the large signal linewidth derived directly from the lasers. Here, we demonstrate a new photonic mmWave platform combining integrated microresonator solitons and high-speed photodiodes to address the challenges in both power and coherence. The solitons, being inherently mode-locked, are measured to provide 5.8 dB additional gain through constructive interference among mmWave beatnotes, and the absolute mmWave power approaches the theoretical limit of conventional heterodyne detection at 100 GHz. In our free-running system, the soliton is capable of reducing the mmWave linewidth by two orders of magnitude from that of the pump laser. Our work leverages microresonator solitons and high-speed modified uni-traveling carrier photodiodes to provide a viable path to chip-scale, high-power, low-noise, high-frequency sources for mmWave applications.

Linear transfer function estimation using the photodiode impulse response.

In this Letter, we implement a model-based auto-regressive estimator of the frequency response for a high-speed photodiode (PD). A transfer function is developed, and the associated coefficients representing the PD model and frequency response are solved for using a two-stage least-squares approach. The model is implemented for a modified uni-traveling carrier PD and experimentally compared to measured impulse response data.

Low dark current III-V on silicon photodiodes by heteroepitaxy.

Top-illuminated PIN and modified uni-traveling carrier (MUTC) photodiodes based on InGaAs/InAlAs/InP were epitaxially grown on Si templates. Photodiodes with 30-µm diameter have dark currents as low as 10 nA at 3 V corresponding to a dark current density of only 0.8 mA/cm2. The responsivity, 3-dB bandwidth, output power and third-order output intercept point (OIP3) were 0.79 A/W, 9 GHz, 2.6 dBm and 15 dBm, respectively.