We demonstrate a method for the fast, high-throughput characterization of the dynamics of active particles. Specifically, we measure the swimming speed distribution and motile cell fraction in Escherichia coli suspensions. By averaging over ∼10(4) cells, our method is highly accurate compared to conventional tracking, yielding a routine tool for motility characterization. We find that the diffusivity of nonmotile cells is enhanced in proportion to the concentration of motile cells.

Sperm require a sense of direction to locate the egg for fertilization. They follow gradients of chemical and physical cues provided by the egg or the oviduct. However, the principles underlying three-dimensional (3D) navigation in chemical landscapes are unknown. Here using holographic microscopy and optochemical techniques, we track sea urchin sperm navigating in 3D chemoattractant gradients. Sperm sense gradients on two timescales, which produces two different steering responses. A periodic component, resulting from the helical swimming, gradually aligns the helix towards the gradient. When incremental path corrections fail and sperm get off course, a sharp turning manoeuvre puts sperm back on track. Turning results from an ‘off' Ca2+ response signifying a chemoattractant stimulation decrease and, thereby, a drop in cyclic GMP concentration and membrane voltage. These findings highlight the computational sophistication by which sperm sample gradients for deterministic klinotaxis. We provide a conceptual and technical framework for studying microswimmers in 3D chemical landscapes.

It is widely believed that the swimming speed, v, of many flagellated bacteria is a nonmonotonic function of the concentration, c, of high-molecular-weight linear polymers in aqueous solution, showing peaked v(c) curves. Pores in the polymer solution were suggested as the explanation. Quantifying this picture led to a theory that predicted peaked v(c) curves. Using high-throughput methods for characterizing motility, we measured v and the angular frequency of cell body rotation, Ω, of motile Escherichia coli as a function of polymer concentration in polyvinylpyrrolidone (PVP) and Ficoll solutions of different molecular weights. We find that nonmonotonic v(c) curves are typically due to low-molecular-weight impurities. After purification by dialysis, the measured v(c) and Ω(c) relations for all but the highest-molecular-weight PVP can be described in detail by Newtonian hydrodynamics. There is clear evidence for non-Newtonian effects in the highest-molecular-weight PVP solution. Calculations suggest that this is due to the fast-rotating flagella seeing a lower viscosity than the cell body, so that flagella can be seen as nano-rheometers for probing the non-Newtonian behavior of high polymer solutions on a molecular scale. T he motility of microorganisms in polymer solutions is a topic of vital biomedical interest. For example, mucus covers the respiratory (1), gastrointestinal (2), and reproductive (3) tracks of all metazoans. Penetration of this solution of biomacromolecules by motile bacterial pathogens is implicated in a range of diseases, e.g., stomach ulcers caused by Helicobacter pylori (4). Oviduct mucus in hens provides a barrier against Salmonella infection of eggs (5). Penetration of the exopolysaccharide matrix of biofilms by swimming bacteria (6) can stabilize or destabilize them in vivo (e.g., the bladder) and in vitro (e.g., catheters). In reproductive medicine (human and veterinary), the motion of sperms in seminal plasma and vaginal mucus, both non-Newtonian polymer solutions, is a strong determinant of fertility (3), and polymeric media are often used to deliver spermicidal and other vaginal drugs (7).Microorganismic propulsion in non-Newtonian media such as high-polymer solutions is also a hot topic in biophysics, soft matter physics, and fluid dynamics (8). Building on knowledge of propulsion modes at low Reynolds number in Newtonian fluids (8), current work seeks to understand how these are modified to enable efficient non-Newtonian swimming. In particular, there is significant interest in a flapping sheet (9, 10) or an undulating filament (11) (modeling the sperm tail) and in a rotating rigid helix (modeling the flagella of, e.g., Escherichia coli) (12, 13) in non-Newtonian fluids.An influential set of experiments in this field was performed 40 years ago by Schneider and Doetsch (SD) (14), who measured the average speed, v, of seven flagellated bacterial species (including E. coli) in solutions of polyvinylpyrrolidone (PVP, molecular weight given as M = 360 kDa) and in methyl cellulose (MC, M...

We present a fast, high-throughput method for characterizing the motility of microorganisms in three dimensions based on standard imaging microscopy. Instead of tracking individual cells, we analyze the spatiotemporal fluctuations of the intensity in the sample from time-lapse images and obtain the intermediate scattering function of the system. We demonstrate our method on two different types of microorganisms: the bacterium Escherichia coli (both smooth swimming and wild type) and the biflagellate alga Chlamydomonas reinhardtii. We validate the methodology using computer simulations and particle tracking. From the intermediate scattering function, we are able to extract the swimming speed distribution, fraction of motile cells, and diffusivity for E. coli, and the swimming speed distribution, and amplitude and frequency of the oscillatory dynamics for C. reinhardtii. In both cases, the motility parameters were averaged over ∼10(4) cells and obtained in a few minutes.

We have studied the dynamics of photo-chemically active colloids made out of silica cores half covered by successive thin layers of Ti and titania, respectively, and moving within aqueous peroxide solutions when exposed to ultraviolet (UV) light. The particles, initially sedimented at the bottom glass wall, exhibit wall-bound states of motion, dependent on the size of the particle, when illuminated from underneath the wall. Upon increasing the intensity of the UV light above a threshold value, which is also dependent on the size of the particle, the particles lift off the wall and move way from it, i.e., they exhibit a photo-gravitactic behavior bearing similarities with that of microorganisms such as phytoplankton and zooplankton. These dependencies on the particle size are rationalized by using a theoretical model of self-phoresis that explicitly accounts for the "shadowing" effect of the Ti/titania layers. This allows us to unequivocally identify the photochemical activity and phototactic response as the key mechanisms beyond the observed phenomenology. Consequently, one has the means to design photo-gravitatic particles that can reversibly switch between operating near a boundary or in the volume away from the boundary by judiciously adjusting the light intensity, i.e., simply by "turning a knob".

Differentiation between distinct stages is fundamental for the life cycle of intracellular protozoan parasites and for transmission between hosts, requiring stringent spatial and temporal regulation. Here, we apply kinome-wide gene deletion and gene tagging in Leishmania mexicana promastigotes to define protein kinases with life cycle transition roles. Whilst 162 are dispensable, 44 protein kinase genes are refractory to deletion in promastigotes and are likely core genes required for parasite replication. Phenotyping of pooled gene deletion mutants using bar-seq and projection pursuit clustering reveal functional phenotypic groups of protein kinases involved in differentiation from metacyclic promastigote to amastigote, growth and survival in macrophages and mice, colonisation of the sand fly and motility. This unbiased interrogation of protein kinase function in Leishmania allows targeted investigation of organelle-associated signalling pathways required for successful intracellular parasitism.

We introduce active, probe-based microrheological techniques for measuring the flow and deformation of complex fluids. These techniques are ideal for mechanical characterization either when little sample is available, or when samples show significant spatial heterogeneity. We review recent results, paying particular attention to comparing and contrasting rheological parameters obtained from micro- and macro-rheological techniques.

We performed passive and active microrheology using probe particles in a bath of well-characterized, model hard-sphere colloids in the fluid state over the whole range of volume fractions below the glass transition. The probe and bath particles have nearly the same size. Passive tracking of probe particles yields short-time self-diffusion coefficients. Comparison with literature data demonstrates that the interaction between probe and bath particles is hard-sphere-like. The short-time diffusivities yield one set of microviscosities as a function of volume fraction, which agrees with previous macrorheological measurements of the high-frequency viscosity of hard-sphere colloids. Using optical tweezers, we measure the force on a trapped probe particle as the rest of the sample is translated at constant velocity. This yields a second set of microviscosities at high Péclet numbers. These agree with previous macrorheological measurements of the high-shear viscosity of similar colloids, at shear-rates below the onset of shear-thickening.