The clock in growing hyphae and their synchronization in Neurospora crassa.

Utilizing a microfluidic chip with serpentine channels, we inoculated the chip with an agar plug with Neurospora crassa mycelium and successfully captured individual hyphae in channels. For the first time, we report the presence of an autonomous clock in hyphae. Fluorescence of a mCherry reporter gene driven by a clock-controlled gene-2 promoter (ccg-2p) was measured simultaneously along hyphae every half an hour for at least 6 days.

CTC-Race: Single-Cell Motility Assay of Circulating Tumor Cells from Metastatic Lung Cancer Patients.

Distinctive subpopulations of circulating tumor cells (CTCs) with increased motility are considered to possess enhanced tumor-initiating potential and contribute to metastasis. Single-cell analysis of the migratory CTCs may increase our understanding of the metastatic process, yet most studies are limited by technical challenges associated with the isolation and characterization of these cells due to their extreme scarcity and heterogeneity. We report a microfluidic method based on CTCs' chemotactic motility, termed as CTC-Race assay, that can analyze migrating CTCs from metastatic non-small-cell lung cancer (NSCLC) patients with advanced tumor stages and enable concurrent biophysical and biochemical characterization of them with single-cell resolution.

Modulation of Dendritic Cell Function via Nanoparticle-Induced Cytosolic Calcium Changes.

Calcium nanoparticles have been investigated for applications, such as drug and gene delivery. Additionally, Ca serves as a crucial second messenger in the activation of immune cells. However, few studies have systematically studied the effects of calcium nanoparticles on the calcium levels and functions within immune cells.

Simultaneous and Multimodal Antigen-Binding Profiling and Isolation of Rare Cells via Quantitative Ferrohydrodynamic Cell Separation.

Simultaneous cell profiling and isolation based on cellular antigen-binding capacity plays an important role in understanding and treating diseases. However, fluorescence-activated cell sorting (FACS) and magnetic-activated cell sorting (MACS) are not able to meet this need, due to their requirement for a large quantity of target cells and the limitation stemming from bimodal separation. Here we report a microfluidic method, termed quantitative ferrohydrodynamic cell separation (qFCS), that achieved multimodal rare cell sorting and simultaneous antigen profiling at a ∼30,000 cell min throughput with a 96.

The macroscopic limit to synchronization of cellular clocks in single cells of Neurospora crassa.

We determined the macroscopic limit for phase synchronization of cellular clocks in an artificial tissue created by a "big chamber" microfluidic device to be about 150,000 cells or less. The dimensions of the microfluidic chamber allowed us to calculate an upper limit on the radius of a hypothesized quorum sensing signal molecule of 13.05 nm using a diffusion approximation for signal travel within the device.

Enhancing membrane-based soft materials with magnetic reconfiguration events.

Adaptive and bioinspired droplet-based materials are built using the droplet interface bilayer (DIB) technique, assembling networks of lipid membranes through adhered microdroplets. The properties of these lipid membranes are linked to the properties of the droplets forming the interface. Consequently, rearranging the relative positions of the droplets within the network will also alter the properties of the lipid membranes formed between them, modifying the transmembrane exchanges between neighboring compartments.

Simultaneous biochemical and functional phenotyping of single circulating tumor cells using ultrahigh throughput and recovery microfluidic devices.

Profiling circulating tumour cells (CTCs) in cancer patients' blood samples is critical to understand the complex and dynamic nature of metastasis. This task is challenged by the fact that CTCs are not only extremely rare in circulation but also highly heterogeneous in their molecular programs and cellular functions. Here we report a combinational approach for the simultaneous biochemical and functional phenotyping of patient-derived CTCs, using an integrated inertial ferrohydrodynamic cell separation (iFCS) method and a single-cell microfluidic migration assay.

Label-free inertial-ferrohydrodynamic cell separation with high throughput and resolution.

Rapid and label-free separation of target cells from biological samples provided unique opportunity for disease diagnostics and treatment. However, even with advanced technologies for cell separation, the limited throughput, high cost and low separation resolution still prevented their utility in separating cells with well-defined physical features from a large volume of biological samples. Here we described an ultrahigh-throughput microfluidic technology, termed as inertial-ferrohydrodynamic cell separation (inertial-FCS), that rapidly sorted through over 60 milliliters of samples at a throughput of 100 000 cells per second in a label-free manner, differentiating the cells based on their physical diameter difference with ∼1-2 μm separation resolution.

Fundamentals of integrated ferrohydrodynamic cell separation in circulating tumor cell isolation.

Methods to separate circulating tumor cells (CTCs) from blood samples were intensively researched in order to understand the metastatic process and develop corresponding clinical assays. However current methods faced challenges that stemmed from CTCs' heterogeneity in their biological markers and physical morphologies. To this end, we developed integrated ferrohydrodynamic cell separation (iFCS), a scheme that separated CTCs independent of their surface antigen expression and physical characteristics.

Brain-on-a-Chip Device for Modeling Multiregional Networks.

Animal models are frequently used in drug discovery because they represent a mammalian in vivo model system, they are the closest approximation to the human brain, and experimentation in humans is not ethical. Working with postmortem human brain samples is challenging and developing human in vitro systems, which mimic the in vivo human brain, has been challenging. However, the use of animal models in drug discovery for human neurological diseases is currently under scrutiny because data from animal models has come with variations due to genetic differences.

Wild Isolates of Reveal Three Conidiophore Architectural Phenotypes.

The vegetative life cycle in the model filamentous fungus, relies on the development of conidiophores to produce new spores. Environmental, temporal, and genetic components of conidiophore development have been well characterized; however, little is known about their morphological variation. We explored conidiophore architectural variation in a natural population using a wild population collection of 21 strains from Louisiana, United States of America (USA).

Label-free ferrohydrodynamic separation of exosome-like nanoparticles.

Isolation of exosomes from biological samples provides a minimally-invasive alternative for basic understanding, diagnosis, and prognosis of metastatic cancers. The biology and clinical values of exosomes are under intensive investigation, yet most studies are limited by technical challenges in recovering these exosomes with heterogeneous sizes and cargos from biological samples. We report a novel method based on "particle ferrohydrodynamics" and its associated microfluidic device, termed as the FerroChip, which can separate exosome-like nanoparticles from microliters of cell culture media and human serum in a label-free, continuous-flow and size-dependent manner, and achieves a high recovery rate (94.

What is Phase in Cellular Clocks?

Four inter-related measures of phase are described to study the phase synchronization of cellular oscillators, and computation of these measures is described and illustrated on single cell fluorescence data from the model filamentous fungus, . One of these four measures is the phase shift in a sinusoid of the form x(t) = cos), where t is time. The other measures arise by creating a replica of the periodic process x(t) called the Hilbert transform which is 90 degrees out of phase with the original process x(t).

Tumor antigen-independent and cell size variation-inclusive enrichment of viable circulating tumor cells.

Isolation of circulating tumor cells (CTCs) from blood provides a minimally-invasive alternative for basic understanding, diagnosis, and prognosis of metastatic cancer. The roles and clinical values of CTCs are under intensive investigation, yet most studies are limited by technical challenges in the comprehensive enrichment of intact and viable CTCs with minimal white blood cell (WBC) contamination. Here, we report a novel method based on contrast of cell magnetization in biocompatible ferrofluids (a colloidal magnetic nanoparticle suspension), termed as integrated ferrohydrodynamic cell separation (iFCS), that enriches CTCs in a tumor antigen-independent and cell size variation-inclusive manner, achieves a high throughput (12 mL h-1), high recovery rate (99.

Manipulation of Single Cells Using a Ferromagnetic Nanorod Cluster Actuated by Weak AC Magnetic Fields.

A unique noncontact single cell manipulation technique based on the actuation of magnetic nanorods (MNRs) or clusters (MCs) by nonuniform alternating magnetic fields (nuAMFs) is demonstrated. Compared to the actuation of MNRs/MCs by conventional magnetophoresis, the motion of MNRs/MCs actuated by nuAMFs can be tuned by additional parameters including the shape of MNRs/MCs and the frequency of the applied magnetic fields. The manipulation of a single cell by an actuated MNR/MC are divided into five stages, i.

Reconfiguring ferromagnetic microrod chains by alternating two orthogonal magnetic fields.

It is well-known that ferromagnetic microrods form linear chains under an external uniform magnetic field B and the chain length is strongly dependent on the applied field, the applied time duration, and the microrod density. When the chains become long enough and the B-field switches to its orthogonal direction, an irreversible morphological transition, i.e.

Microfluidics in Malignant Glioma Research and Precision Medicine.

Glioblastoma multiforme (GBM) is an aggressive form of brain cancer that has no effective treatments and a prognosis of only 12-15 months. Microfluidic technologies deliver microscale control of fluids and cells, and have aided cancer therapy as point-of-care devices for the diagnosis of breast and prostate cancers. However, a few microfluidic devices are developed to study malignant glioma.

Magnetic resonance conditional paramagnetic choke for suppression of imaging artifacts during magnetic resonance imaging.

Higher risk patient populations require continuous physiological monitoring and, in some cases, connected life-support systems, during magnetic resonance imaging examinations. While recently there has been a shift toward wireless technology, some of the magnetic resonance imaging devices are still connected to the outside using cabling that could interfere with the magnetic resonance imaging's radio frequency during scanning, resulting in excessive heating. We developed a passive method for radio frequency suppression on cabling that may assist in making some of these devices magnetic resonance imaging compatible.

Ferrofluid-Based Droplet Interface Bilayer Networks.

Droplet interface bilayer (DIB) networks allow for the construction of stimuli-responsive, membrane-based materials. Traditionally used for studying cellular transport phenomena, the DIB technique has proven its practicality when creating structured droplet networks. These structures consist of aqueous compartments capable of exchanging their contents across membranous barriers in a regulated fashion via embedded biomolecules, thus approximating the activity of natural cellular systems.

Label-free ferrohydrodynamic cell separation of circulating tumor cells.

Circulating tumor cells (CTCs) have significant implications in both basic cancer research and clinical applications. To address the limited availability of viable CTCs for fundamental and clinical investigations, effective separation of extremely rare CTCs from blood is critical. Ferrohydrodynamic cell separation (FCS), a label-free method that conducted cell sorting based on cell size difference in biocompatible ferrofluids, has thus far not been able to enrich low-concentration CTCs from cancer patients' blood because of technical challenges associated with processing clinical samples.

Label-Free Microfluidic Manipulation of Particles and Cells in Magnetic Liquids.

Manipulating particles and cells in magnetic liquids through so-called "negative magnetophoresis" is a new research field. It has resulted in label-free and low-cost manipulation techniques in microfluidic systems and many exciting applications. It is the goal of this review to introduce the fundamental principles of negative magnetophoresis and its recent applications in microfluidic manipulation of particles and cells.

Biocompatible and label-free separation of cancer cells from cell culture lines from white blood cells in ferrofluids.

This paper reports a biocompatible and label-free cell separation method using ferrofluids that can separate a variety of low-concentration cancer cells from cell culture lines (∼100 cancer cells per mL) from undiluted white blood cells, with a throughput of 1.2 mL h and an average separation efficiency of 82.2%.

Glioma Cell Invasion is Significantly Enhanced in Composite Hydrogel Matrices Composed of Chondroitin 4- and 4,6-Sulfated Glycosaminoglycans.

Glioblastoma multiforme (GBM) is the most aggressive form of astrocytoma accounting for a majority of primary malignant brain tumors in the United States. Chondroitin sulfate proteoglycans (CSPGs) and their glycosaminoglycan (GAG) side chains are key constituents of the brain extracellular matrix (ECM) implicated in promoting tumor invasion. However, the mechanisms by which sulfated CS-GAGs promote brain tumor invasion are currently unknown.

Synchronizing stochastic circadian oscillators in single cells of Neurospora crassa.

The synchronization of stochastic coupled oscillators is a central problem in physics and an emerging problem in biology, particularly in the context of circadian rhythms. Most measurements on the biological clock are made at the macroscopic level of millions of cells. Here measurements are made on the oscillators in single cells of the model fungal system, Neurospora crassa, with droplet microfluidics and the use of a fluorescent recorder hooked up to a promoter on a clock controlled gene-2 (ccg-2).