Two-dimensional planar swimming selects for high DNA integrity sperm.

Selection of high-quality sperm is critical to the success of assisted reproductive technologies. Clinical screening for top sperm has long focused on sperm swimming ability when following boundaries or when fully free of constraints. In this work, we demonstrate a sperm selection approach with parallel 2 µm tall confined selection channels that prohibit rotation of the sperm head and require planar swimming. We demonstrate that a planar swimming subpopulation of sperm capable of entering and navigating these channels has DNA integrity superior to the freely-swimming motile or raw sperm populations over a wide range of semen sample qualities. The DNA integrity of the selected sperm was significantly higher than that of the corresponding raw samples for donor samples and clinical patient samples, respectively. In side-by-side testing, this method outperforms current clinical selection methods, density gradient centrifugation and swim-up, as well as sperm selected via general motility. Planar swimming represents a viable sperm selection mechanism with the potential to improve outcomes for couples and offspring.

A Platform for High-Throughput Assessments of Environmental Multistressors.

A platform compatible with microtiter plates to parallelize environmental treatments to test the complex impacts of multiple stressors, including parameters relevant to climate change and point source pollutants is developed. This platform leverages (1) the high rate of purely diffusive gas transport in aerogels to produce well-defined centimeter-scale gas concentration gradients, (2) spatial light control, and (3) established automated liquid handling. The parallel gaseous, aqueous, and light control provided by the platform is compatible with multiparameter experiments across the life sciences. The platform is applied to measure biological effects in over 700 treatments in a five-parameter full factorial study with the microalgae Chlamydomonas reinhardtii. Further, the CO2 response of multicellular organisms, Lemna gibba and Artemia salina under surfactant and nanomaterial stress are tested with the platform.

Microfluidics for sperm analysis and selection.

Infertility is a growing global health issue with far-reaching socioeconomic implications. A downward trend in male fertility highlights the acute need for affordable and accessible diagnosis and treatment. Assisted reproductive technologies are effective in treating male infertility, but their success rate has plateaued at ∼33% per cycle. Many emerging opportunities exist for microfluidics - a mature technology in other biomedical areas - in male infertility diagnosis and treatment, and promising microfluidic approaches are under investigation for addressing male infertility. Microfluidic approaches can improve our fundamental understanding of sperm motion, and developments in microfluidic devices that use microfabrication and sperm behaviour can aid semen analysis and sperm selection. Many burgeoning possibilities exist for engineers, biologists, and clinicians to improve current practices for infertility diagnosis and treatment. The most promising avenues have the potential to improve medical practice, moving innovations from research laboratories to clinics and patients in the near future.

A penalty on photosynthetic growth in fluctuating light.

Fluctuating light is the norm for photosynthetic organisms, with a wide range of frequencies (0.00001 to 10 Hz) owing to diurnal cycles, cloud cover, canopy shifting and mixing; with broad implications for climate change, agriculture and bioproduct production. Photosynthetic growth in fluctuating light is generally considered to improve with increasing fluctuation frequency. Here we demonstrate that the regulation of photosynthesis imposes a penalty on growth in fluctuating light for frequencies in the range of 0.01 to 0.1 Hz (organisms studied: Synechococcus elongatus and Chlamydomonas reinhardtii). We provide a comprehensive sweep of frequencies and duty cycles. In addition, we develop a 2nd order model that identifies the source of the penalty to be the regulation of the Calvin cycle - present at all frequencies but compensated at high frequencies by slow kinetics of RuBisCO.

Light dilution via wavelength management for efficient high-density photobioreactors.

The spectral distribution of light influences microalgae productivity; however, development of photobioreactors has proceeded largely without regard to spectral optimization. Here, we use monochromatic light to quantify the joint influence of path length, culture density, light intensity, and wavelength on productivity and efficiency in Synechococcus elongatus. The productivity of green light was ∼4× that of red at the highest levels of culture density, depth, and light intensity. This performance is attributed to the combination of increased dilution and penetration of this weakly absorbed wavelength over a larger volume fraction of the reactor. In contrast, red light outperformed other wavelengths in low-density cultures with low light intensities. Low-density cultures also adapted more rapidly to reduce absorption of longer wavelengths, allowing for prolonged cultivation. Taken together, these results indicate that, particularly for artificially lit photobioreactors, wavelength needs to be included as a critical operational parameter to maintain optimal performance. Biotechnol. Bioeng. 2017;114: 1160-1169. © 2017 Wiley Periodicals, Inc.

Dual gradients of light intensity and nutrient concentration for full-factorial mapping of photosynthetic productivity.

Optimizing bioproduct generation from microalgae is complicated by the myriad of coupled parameters affecting photosynthetic productivity. Quantifying the effect of multiple coupled parameters in full-factorial fashion requires a prohibitively high number of experiments. We present a simple hydrogel-based platform for the rapid, full-factorial mapping of light and nutrient availability on the growth and lipid accumulation of microalgae. We accomplish this without microfabrication using thin sheets of cell-laden hydrogels. By immobilizing the algae in a hydrogel matrix we are able to take full advantage of the continuous spatial chemical gradient produced by a diffusion-based gradient generator while eliminating the need for chambers. We map the effect of light intensities between 0 µmol m(-2) s(-1) and 130 µmol m(-2) s(-1) (∼28 W m(-2)) coupled with ammonium concentrations between 0 mM and 7 mM on Chlamydomonas reinhardtii. Our data set, verified with bulk experiments, clarifies the role of ammonium availability on the photosynthetic productivity Chlamydomonas reinhardtii, demonstrating the dependence of ammonium inhibition on light intensity. Specifically, a sharp optimal growth peak emerges at approximately 2 mM only for light intensities between 80 and 100 µmol m(-2) s(-1)- suggesting that ammonium inhibition is insignificant at lower light intensities. We speculate that this phenomenon is due to the regulation of the high affinity ammonium transport system in Chlamydomonas reinhardtii as well as free ammonia toxicity. The complexity of this photosynthetic biological response highlights the importance of full-factorial data sets as enabled here.

Predominance of sperm motion in corners.

Sperm migration through the female tract is crucial to fertilization, but the role of the complex and confined structure of the fallopian tube in sperm guidance remains unknown. Here, by confocal imaging microchannels head-on, we distinguish corner- vs. wall- vs. bulk-swimming bull sperm in confined geometries. Corner-swimming dominates with local areal concentrations as high as 200-fold that of the bulk. The relative degree of corner-swimming is strongest in small channels, decreases with increasing channel size, and plateaus for channels above 200 µm. Corner-swimming remains predominant across the physiologically-relevant range of viscosity and pH. Together, boundary-following sperm account for over 95% of the sperm distribution in small rectangular channels, which is similar to the percentage of wall swimmers in circular channels of similar size. We also demonstrate that wall-swimming sperm travel closer to walls in smaller channels (~100 µm), where the opposite wall is within the hydrodynamic interaction length-scale. The corner accumulation effect is more than the superposition of the influence of two walls, and over 5-fold stronger than that of a single wall. These findings suggest that folds and corners are dominant in sperm migration in the narrow (sub-mm) lumen of the fallopian tube and microchannel-based sperm selection devices.

Microalgae on display: a microfluidic pixel-based irradiance assay for photosynthetic growth.

Microalgal biofuel is an emerging sustainable energy resource. Photosynthetic growth is heavily dependent on irradiance, therefore photobioreactor design optimization requires comprehensive screening of irradiance variables, such as intensity, time variance and spectral composition. Here we present a microfluidic irradiance assay which leverages liquid crystal display technology to provide multiplexed screening of irradiance conditions on growth. An array of 238 microreactors are operated in parallel with identical chemical environments. The approach is demonstrated by performing three irradiance assays. The first assay evaluates the effect of intensity on growth, quantifying saturating intensity. The second assay quantifies the influence of time-varied intensity and the threshold frequency for growth. Lastly, the coupled influence of red-blue spectral composition and intensity is assessed. Each multiplexed assay is completed within three days. In contrast, completing the same number of experiments using conventional incubation flasks would require several years. Not only does our approach enable more rapid screening, but the short optical path avoids self-shading issues inherent to flask based systems.

Induced detachment of coalescing droplets on superhydrophobic surfaces.

Coalescence of a falling droplet with a stationary sessile droplet on a superhydrophobic surface is investigated by a combined experimental and numerical study. In the experiments, the droplet diameter, the impact velocity, and the distance between the impacting droplets were controlled. The evolution of surface shape during the coalescence of two droplets on the superhydrophobic surface is captured using high speed imaging and compared with numerical results. A two-phase volume of fluid (VOF) method is used to determine the dynamics of droplet coalescence, shape evaluation, and contact line movement. The spread length of two coalesced droplets along their original center is also predicted by the model and compared well with the experimental results. The effect of different parameters such as impact velocity, center to center distance, and droplet size on contact time and restitution coefficient are studied and compared to the experimental results. Finally, the wetting and the self-cleaning properties of superhydrophobic surfaces have been investigated. It has been found that impinging water drops with very small amount of kinetic impact energy were able to thoroughly clean these surfaces.