We demonstrate a new computational illumination technique that achieves large space-bandwidth-time product, for quantitative phase imaging of unstained live samples in vitro. Microscope lenses can have either large field of view (FOV) or high resolution, not both. Fourier ptychographic microscopy (FPM) is a new computational imaging technique that circumvents this limit by fusing information from multiple images taken with different illumination angles. The result is a gigapixelscale image having both wide FOV and high resolution, i.e. large space-bandwidth product (SBP). FPM has enormous potential for revolutionizing microscopy and has already found application in digital pathology. However, it suffers from long acquisition times (on the order of minutes), limiting throughput. Faster capture times would not only improve imaging speed, but also allow studies of live samples, where motion artifacts degrade results. In contrast to fixed (e.g. pathology) slides, live samples are continuously evolving at various spatial and temporal scales. Here, we present a new source coding scheme, along with real-time hardware control, to achieve 0.8 NA resolution across a 4× FOV with sub-second capture times. We propose an improved algorithm and new initialization scheme, which allow robust phase reconstruction over long time-lapse experiments. We present the first FPM results for both growing and confluent in vitro cell cultures, capturing videos of subcellular dynamical phenomena in popular cell lines undergoing division and migration. Our method opens up FPM to applications with live samples, for observing rare events in both space and time.

Abstract. We demonstrate a single-camera imaging system that can simultaneously acquire brightfield, darkfield, and phase contrast images in real time. Our method uses computational illumination via a programmable light-emitting diode (LED) array at the source plane, providing flexible patterning of illumination angles. Brightfield, darkfield, and differential phase contrast images are obtained by changing the LED patterns, without any moving parts. Previous work with LED array illumination was only valid for static samples because the hardware speed was not fast enough to meet real-time acquisition and processing requirements. Here, we time multiplex patterns for each of the three contrast modes in order to image dynamic biological processes in all three contrast modes simultaneously. We demonstrate multicontrast operation at the maximum frame rate of our camera (50 Hz with 2160 × 2560 pixels).

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most important wheat diseases worldwide in areas with cool or maritime climates. Wild emmer (Triticum turgidum var. dicoccoides) is an important potential donor of disease resistances and other traits for common wheat improvement. A powdery mildew resistance gene was transferred from wild emmer accession G-303-1M to susceptible common wheat by crossing and backcrossing, resulting in inbred line P63 (Yanda1817/G-303-1 M//3*Jing411, BC(2)F(6)). Genetic analysis of an F(2) population and the F(2:3) families developed from a cross of P63 and a susceptible common wheat line Xuezao showed that the powdery mildew resistance in P63 was controlled by a single recessive gene. Molecular markers and bulked segregant analysis were used to characterize and map the powdery mildew resistance gene. Nine genomic SSR markers (Xbarc7, Xbarc55, Xgwm148, Xgwm257, Xwmc35, Xwmc154, Xwmc257, Xwmc382, Xwmc477), five AFLP-derived SCAR markers (XcauG3, XcauG6, XcauG10, XcauG20, XcauG22), three EST-STS markers (BQ160080, BQ160588, BF146221) and one RFLP-derived STS marker (Xcau516) were linked to the resistance gene, designated pm42, in P63. pm42 was physically mapped on chromosome 2BS bin 0.75-0.84 using Chinese Spring nullisomic-tetrasomic, ditelosomic and deletion lines, and was estimated to be more than 30 cM proximal to Xcau516, a RFLP-derived STS marker that co-segregated with the wild emmer-derived Pm26 which should be physically located in 2BS distal bin 0.84-1.00. pm42 was highly effective against 18 of 21 differential Chinese isolates of B. graminis f. sp. tritici. The closely linked molecular markers will enable the rapid transfer of pm42 to wheat breeding populations thus adding to their genetic diversity.

Bitter gourd (Momordica charantia) is a popular cultivated vegetable in Asian and African countries. To reveal the characteristics of the genomic structure, evolutionary trajectory, and genetic basis underlying the domestication of bitter gourd, we performed whole-genome sequencing of the cultivar Dali-11 and the wild small-fruited line TR and resequencing of 187 bitter gourd germplasms from 16 countries. The major gene clusters (Bi clusters) for the biosynthesis of cucurbitane triterpenoids, which confer a bitter taste, are highly conserved in cucumber, melon, and watermelon. Comparative analysis among cucurbit genomes revealed that the Bi cluster involved in cucurbitane triterpenoid biosynthesis is absent in bitter gourd. Phylogenetic analysis revealed that the TR group, including 21 bitter gourd germplasms, may belong to a new species or subspecies independent from M. charantia. Furthermore, we found that the remaining 166 M. charantia germplasms are geographically differentiated, and we identified 710, 412, and 290 candidate domestication genes in the South Asia, Southeast Asia, and China populations, respectively. This study provides new insights into bitter gourd genetic diversity and domestication and will facilitate the future genomics-enabled improvement of bitter gourd.