Cell surface receptors: rapid quantification of live cell receptors using bioluminescence in a flow-based microfluidic device (small 8/2015).

C. T. Lim and co-workers describe a rapid and sensitive bioluminescence-based microfluidic method for quantifying receptor numbers on live cells.

Rapid quantification of live cell receptors using bioluminescence in a flow-based microfluidic device.

The number of receptors expressed by cells plays an important role in controlling cell signaling events, thus determining its behaviour, state and fate. Current methods of quantifying receptors on cells are either laborious or do not maintain the cells in their native form. Here, a method integrating highly sensitive bioluminescence, high precision microfluidics and small footprint of lensfree optics is developed to quantify cell surface receptors.

Microfluidic platform for negative enrichment of circulating tumor cells.

Negative enrichment is the preferred approach for tumor cell isolation as it does not rely on biomarker expression. However, size-based negative enrichment methods suffer from well-known recovery/purity trade-off. Non-size based methods have a number of processing steps that lead to compounded cell loss due to extensive sample processing and handling which result in a low recovery efficiency.

Electrokinetic Analysis of Cell Translocation in Low-Cost Microfluidic Cytometry for Tumor Cell Detection and Enumeration.

Conventional Coulter counters have been introduced as an important tool in biological cell assays since several decades ago. Recently, the emerging portable Coulter counter has demonstrated its merits in point of care diagnostics, such as on chip detection and enumeration of circulating tumor cells (CTC). The working principle is based on the cell translocation time and amplitude of electrical current change that the cell induces.

Size based sorting and patterning of microbeads by evaporation driven flow in a 3D micro-traps array.

We present a three-dimensional (3D) micro-traps array for size selective sorting and patterning of microbeads via evaporation-driven capillary flow. The interconnected micro-traps array was manufactured by silicon micromachining. Microliters of aqueous solution containing particle mixtures of different sized (0.

Effects of the electrode size and modification protocol on a label-free electrochemical biosensor.

In the present work, the effect of a surface modification protocol along with the electrode size has been investigated for developing an efficient, label-free electrochemical biosensing method for diagnosis of traumatic brain injury (TBI) biomarkers. A microdisk electrode array (MDEA) and a macroelectrode with a comb structure (MECS) were modified with an anti-GFAP (GFAP = glial fibrillary acidic protein) antibody using two protocols for optimum and label-free detection of GFAP, a promising acute-phase TBI biomarker. For the MDEA, an array of six microdisks with a 100 μm diameter and, for the MECS, a 3.

Enrichment, detection and clinical significance of circulating tumor cells.

Circulating Tumor Cells (CTCs) are shed from primary or secondary tumors into blood circulation. Accessing and analyzing these cells provides a non-invasive alternative to tissue biopsy. CTCs are estimated to be as few as 1 cell among a few million WBCs and few billion RBCs in 1 ml of patient blood and are rarely found in healthy individuals.

Towards an optimal and unbiased approach for tumor cell isolation.

Our current understanding of clinical significance or the lack thereof of circulating tumor cells (CTCs) is biased by the technology used to isolate these rare cells. Despite the presence of a vast number of academic and commercial technologies, the lack of a standardized and optimized platform has been widely noted. We present a negative enrichment approach, integrating WBC depletion and chemical-free RBC depletion in the same setup without the need for centrifugation, washing or multiple sample handling steps.

3D numerical simulation of a Coulter counter array with analysis of electrokinetic forces.

Coulter counters have played an important role in biological cell assays since their introduction decades ago. Several types of high throughput micro-Coulter counters based on lab-on-chip devices have been commercialized recently. In this paper, we propose a highly integrated micro-Coulter counter array working under low DC voltage.

Anti-EpCAM modified LC-SPDP monolayer on gold microelectrode based electrochemical biosensor for MCF-7 cells detection.

A succinimidyl 6-(3-[2-pyridyldithio]-propionamido) hexanoate (LC-SPDP) self-assembled monolayer (SAM) prepared onto a 500 μm (diameter) gold microelectrode (Au) surface has been utilized for covalent immobilization of anti-EpCAM antibody. Amino group on anti-EpCAM antibody was covalently bound with succinimidyl group on SAM via amide bond and unreacted active groups of LC-SPDP were blocked using 1% ethanol amine (EA). These anti-EpCAM/LC-SPDP/Au electrodes were characterized using cyclic voltammetric (CV) and fluorescence techniques, respectively.

Breast tumor cell detection at single cell resolution using an electrochemical impedance technique.

Gold micro-electrodes with various diameters (25, 50, 75, 100 and 250 μm) were manufactured using standard micro-fabrication techniques and optimized for counting of MCF-7 cells (breast tumor cells) with single cell resolution. For specific cell capture, anti-EpCAM was immobilized on 11-mercaptoundecanoic acid (11-MUA)-3-mercaptopropionic acid (3-MPA) mixed self-assembled monolayer (SAM) modified gold surface of micro-electrodes. Electrodes were characterized using optical, cyclic voltammetry and electrochemical impedance spectroscopic (EIS) techniques.

A detailed model for high-frequency impedance characterization of ovarian cancer epithelial cell layer using ECIS electrodes.

We report on the electrical impedance spectroscopy characterization of OVCA429 ovarian cancer cells. A commercially available eight-well cell culture impedance array (ECIS-8W1E), commonly used in electrical cell-substrate impedance sensing (ECIS), was used for OVCA429 characterization. Impedances of ECIS-8W1E array were recorded with cell culture medium (without cells) and with OVCA429 cell layer in the culture medium between 100 Hz and 10 MHz frequency.

Design considerations in the development and application of microdisc electrode arrays (MDEAs) for implantable biosensors.

The use of microlithographically fabricated Microdisc Electrode Arrays (MDEAs) in the development of implantable voltammetric biosensors necessitates design criteria that balances the overall footprint of the device with the advantages to be derived from large separation distances between non-interacting microdisc elements. Using the dynamic electroanalytical techniques of Multiple Scan Rate Cyclic Voltammetry (MSRCV) experiments with finite element simulations and Electrochemical Impedance Spectroscopy with equivalent circuit modeling, three unique MDEA designs; MDEA 050 (r = 25 microm, 5,184 discs), MDEA 100 (r = 50 microm, 1,296 discs) and MDEA 250 (r = 125 microm, 207 discs) of constant critical dimensions (center-to-center d/r = 4) and area (A = 0.1 cm(2)) were studied in 1.

Design rule for optimization of microelectrodes used in electric cell-substrate impedance sensing (ECIS).

This paper presents an experimentally derived design rule for optimization of microelectrodes used in electric cell-substrate impedance sensing (ECIS) up to 10MHz. The effect of change in electrode design (through electrode sensor area, lead trace widths, and passivation coating thickness) on electrode characteristics was experimentally evaluated using electrochemical impedance spectroscopy (EIS) measurements and analyzed using equivalent circuit models. A parasitic passivation coating impedance was successfully minimized by designing electrodes with either a thicker passivation layer or a smaller lead trace area.

Biomimetic hydrogels for biosensor implant biocompatibility: electrochemical characterization using micro-disc electrode arrays (MDEAs).

Our interest is in the development of engineered microdevices for continuous remote monitoring of intramuscular lactate, glucose, pH and temperature during post-traumatic hemorrhaging. Two important design considerations in the development of such devices for in vivo diagnostics are discussed; the utility of micro-disc electrode arrays (MDEAs) for electrochemical biosensing and the application of biomimetic, bioactive poly(HEMA)-based hydrogel composites for implant biocompatibility. A poly(HEMA)-based hydrogel membrane containing polyethylene glycol (PEG) was UV cross-linked with tetraethyleneglycol diacrylate following application to MDEAs (50 mum discs) and to 250 mum diameter gold electrodes within 8-well culture ware.

Towards an implantable biochip for glucose and lactate monitoring using microdisc electrode arrays (MDEAs).

A complete electrochemical cell-on-a-chip that uses the MicroDisc Electrode Array (MDEA) working electrode (WE) design was evaluated for eventual intramuscular implantation for the continuous amperometric monitoring of glucose and lactate in an animal trauma model. The microfabricated ECC MDEA5037 comprises two discrete electrochemical cells-on-a-chip (ECCs), each with a reference, counter, and MDEA working electrode. Each MDEA comprises 37 microdiscs (diameter = 50 microm) arranged in a Hexagonal Closed Packed (HCP) arrangement with a center to center distance (d) of 100 microm.