Clusters of circulating tumor cells traverse capillary-sized vessels

Multicellular aggregates of circulating tumor cells (CTC clusters) are potent initiators of distant organ metastasis. However, it is currently assumed that CTC clusters are too large to pass through narrow vessels to reach these organs. Here, we present evidence that challenges this assumption through the use of microfluidic devices designed to mimic human capillary constrictions and CTC clusters obtained from patient and cancer cell origins. Over 90% of clusters containing up to 20 cells successfully traversed 5- to 10-μm constrictions even in whole blood. Clusters rapidly and reversibly reorganized into single-file chain-like geometries that substantially reduced their hydrodynamic resistances. Xenotransplantation of human CTC clusters into zebrafish showed similar reorganization and transit through capillary-sized vessels in vivo. Preliminary experiments demonstrated that clusters could be disrupted during transit using drugs that affected cellular interaction energies. These findings suggest that CTC clusters may contribute a greater role to tumor dissemination than previously believed and may point to strategies for combating CTC cluster-initiated metastasis

Circulating tumor cells detected by lab-on-adisc: Role in early diagnosis of gastric cancer

[Background] The use of circulating tumor cells (CTCs) as an early diagnostic biomarker and prognostic indicator after surgery or chemotherapy has been suggested for various cancers. This study aimed to evaluate CTCs in patients who underwent gastrectomy for gastric cancer and to explore their clinical usefulness in the early diagnosis of gastric cancer. [Methods] A total of 116 patients with gastric cancer who underwent gastrectomy and 31 healthy volunteers were prospectively included between 2014 and 2015. Peripheral blood samples were collected before gastrectomy, and CTCs were examined using a centrifugal microfluidic system with a new fluid-assisted separation technique. [Results] After creating a receiver operating characteristic curve to identify the discriminative CTC value needed differentiate patients with gastric cancer from healthy volunteers, sensitivity and specificity were nearly optimized at a CTC threshold of 2 per 7.5 mL of blood. Of the 102 persons with a CTC level >= 2 per 7.5 mL of blood, 99 (97.1%) had gastric cancer, and of the 45 persons with a CTC level <2 per 7.5 mL of blood, 28 (62.2%) were healthy controls. Accordingly, the sensitivity and specificity for the differentiation of patients with gastric cancer from healthy controls were 85.3% and 90.3%, respectively. However, the presence of CTCs was not associated with any clinicopathologic features such as staging, histologic type, or mucin phenotype. [Conclusion] Although we could not prove the clinical feasibility of CTCs for gastric cancer staging, our results suggest a potential role of CTCs as an early diagnostic biomarker of gastric cancer

Characterization of cross-coupling in capacitive micromachined ultrasonic transducers

This paper analyzes element-to-element and cell-to-cell cross-coupling in capacitive micromachined ultrasonic transducers (cMUTs) using an interferometer. In a 1-D linear cMUT array immersed in oil, a single element was excited, and membrane displacements were measured at different positions along the array with an interferometer. Electrical measurements of the received voltage on each array element were also performed simultaneously to verify the optical measurements. The array was then covered with a polydimethylsiloxane (PDMS) layer, and the cross-coupling measurements were repeated. The cross-coupling levels for conventional and collapsed operation of the cMUT were compared. Since the cMUTs were immersed in oil, the optical measurements were corrected for acousto-optic interaction, and the results were reviewed in time-spatial and frequency-spatial domains. The main cross-coupling mechanism was due to the dispersive guided modes supported by the membrane periodicity. In both modes of operation, cross-coupling dispersion curves predicted a gradual reduction in phase velocity at higher frequencies. At lower frequencies, this phase velocity tended to approach 1480 m/s asymptotically. Better cross-coupling suppression was observed in the collapsed (-34 dB) than the conventional operation (-23 dB). The element-to-element cross-coupling experiments showed that a 5-mu m PDMS layer reduced the measured cross-coupling levels down to -39 dB in the collapsed operatio

The emergence of multifrequency force microscopy

Atomic force microscopy uses the deflection of a cantilever with a sharp tip to examine surfaces, and conventional dynamic force microscopy involves the excitation and detection of a single frequency component of the tip’s motion. Information about the properties of a sample is, however, encoded in the motion of the probe and the dynamics of the cantilever are highly nonlinear. Therefore, information included in the other frequency components is irreversibly lost. Multifrequency force microscopy involves the excitation and/or detection of several frequencies of the probe’s oscillation, and has the potential to overcome limitations in spatial resolution and acquisition times of conventional force microscopes. It could also provide new applications in fields such as energy storage and nanomedicine. Here we review the development of multifrequency force microscopy methods, highlighting the five most prominent approaches. We also examine the range of applications offered by the technique, which include mapping the flexibility of proteins, imaging the mechanical vibrations of carbonbased resonators, mapping ion diffusion, and imaging the subsurface of cells.We are grateful for financial support from the Ministerio de Ciencia e Innovación (CSD2010-00024, MAT2009-08650).Peer reviewe

Label-free enumeration, collection and downstream cytological and cytogenetic analysis of circulating tumor cells

Circulating tumor cells (CTCs) have a great potential as indicators of metastatic disease that may help physicians improve cancer prognostication, treatment and patient outcomes. Heterogeneous marker expression as well as the complexity of current antibody-based isolation and analysis systems highlights the need for alternative methods. In this work, we use a microfluidic Vortex device that can selectively isolate potential tumor cells from blood independent of cell surface expression. This system was adapted to interface with three protein-marker-free analysis techniques: (i) an in-flow automated image processing system to enumerate cells released, (ii) cytological analysis using Papanicolaou (Pap) staining and (iii) fluorescence in situ hybridization (FISH) targeting the ALK rearrangement. In-flow counting enables a rapid assessment of the cancer-associated large circulating cells in a sample within minutes to determine whether standard downstream assays such as cytological and cytogenetic analyses that are more time consuming and costly are warranted. Using our platform integrated with these workflows, we analyzed 32 non-small cell lung cancer (NSCLC) and 22 breast cancer patient samples, yielding 60 to 100% of the cancer patients with a cell count over the healthy threshold, depending on the detection method used: respectively 77.8% for automated, 60–100% for cytology, and 80% for immunostaining based enumeration