Cell sorting actuated by a microfluidic inertial vortex.

The sorting of specific cell populations is an established tool in biological research, with new applications demanding greater cell throughput, sterility and elimination of cross-contamination. Here we report 'vortex-actuated cell sorting' (VACS), a new technique that deflects cells individually, via the generation of a transient microfluidic vortex by a thermal vapour bubble: a novel mechanism, which is able to sort cells based on fluorescently-labelled molecular markers. Using in silico simulation and experiments on beads, an immortal cell line and human peripheral blood mononuclear cells (PBMCs), we demonstrate high-purity and high-recovery sorting with input rates up to 104 cells per s and switching speeds comparable to existing techniques (>40 kHz). A tiny footprint (1 × 0.25 mm) affords miniaturization and the potential to achieve multiplexing: a crucial step in increasing processing rate. Simple construction using biocompatible materials potentially minimizes cost of fabrication and permits single-use sterile cartridges. We believe VACS potentially enables parallel sorting at throughputs relevant to cell therapy, liquid biopsy and phenotypic screening.

A self-mixing laser-diode interferometer for measuring basilar membrane vibrations without opening the cochlea.

A laser-diode forms the basis of a displacement sensitive homodyne interferometer suitable for measurements from poorly reflective surfaces. The compact and cost-effective interferometer utilizes the self-mixing effect when laser light reflected from a moving target re-enters the laser cavity and causes phase dependent changes of the lasing intensity. A piezo positioner was used to displace the interferometer with known frequency and amplitude as a basis for real-time calibration of the interferometer's sensitivity. The signal-processing algorithm is described that allows measurements in presence of high amplitude noise leading to variation of the interferometer's operating point. Measurements of sound-induced basilar membrane displacements were made in the intact cochleae of rodents by focusing the laser beam of the interferometer through the transparent round window membrane. The interferometer provides a viable means for making subnanometre mechanical measurements from structures in the inner ears of small mammals, where opening of the cochlea is not practicable.

The mechanical properties of chick (Gallus domesticus) sensory hair bundles: relative contributions of structures sensitive to calcium chelation and subtilisin treatment.

Up to four link types are found between the stereocilia of chick vestibular hair bundles: tip links, horizontal top connectors, shaft connectors and ankle links. A fifth type, the kinocilial link, couples the hair bundle to the kinocilium. Brownian-motion microinterferometry was used to study the mechanical properties of the hair bundle and investigate changes caused by removing different links with the calcium chelator BAPTA or the protease subtilisin. Immunofluorescence with an antibody to the hair-cell antigen (HCA) and electron microscopy were used to verify destruction of the links. The root mean square displacement and the corresponding absolute stiffness of untreated hair bundles were 4.3 nm and 0.9 mN m(-1), respectively. The ratio of Brownian-motion spectra before and after treatment was calculated and processed using a single oscillator model to obtain relative stiffness. Treatment with BAPTA, which cleaves tip, kinocilial and ankle links, reduces hair-bundle stiffness by 43%, whilst subtilisin treatment, which breaks ankle links and shaft connectors, reduces stiffness by 48%. No changes were detected in viscous damping following either treatment. The time course of the subtilisin-induced stiffness change was close to that of HCA loss, but not to the disappearance of the ankle links, suggesting that shaft connectors make a more significant contribution to hair-bundle stiffness. Sequential treatments of the hair bundles with BAPTA and subtilisin show that the effects are additive. The implication of complete additivity is that structures resistant to both agents (e.g. top connectors and stereocilia pivots) are responsible for approximately 9% of the overall bundle stiffness.