Jae Chan Park

Surface acoustic wave immunosensor for real-time detection of hepatitis B surface antibodies in whole blood samples

We demonstrate an application of Love wave mode surface acoustic wave (SAW) immunosensor to detect hepatitis B surface antibody (HBsAb) in aqueous conditions. SiO(2) guiding layer was deposited on 36 degrees YX-LiTaO(3) piezoelectric single crystal substrate to protect the electrodes and to trap the acoustic energy near the surface, and hepatitis B surface antigen (HBsAg) was immobilized on the sensing area. The resonance frequency shift was monitored to detect specific binding of HBsAb to immobilized HBsAg. To eliminate the effects of other physical factors except for the mass change, the resonance frequency was compared to that of a reference SAW device coated with bovine serum albumin (BSA) to block binding of HBsAb. The guiding layer thickness with maximum mass sensitivity was found to be 5 microm, which was in agreement with the theoretical calculation, and the center resonance frequency was around 199 MHz. The sensor showed binding specificity to HBsAb and a linear relationship between the frequency shift and the antibody concentration with sensitivity of 0.74 Hz/(pg/microl) and detection limit below 10 pg/microl. In addition, our SAW immunosensor successfully detected HBsAb in whole blood samples without any pretreatment, opening up its applicability in fast label-free protein detection methods.

Sensitive and simultaneous detection of cardiac markers in human serum using surface acoustic wave immunosensor.

We present a rapid and sensitive surface acoustic wave (SAW) immunosensor that utilizes gold staining as a signal enhancement method. A sandwich immunoassay was performed on sensing area of the SAW sensor, which could specifically capture and detect cardiac markers (cardiac troponin I (cTnI), creatine kinase (CK)-MB, and myoglobin). The analytes in human serum were captured on gold nanoparticles (AuNPs) that were conjugated in advance with detection antibodies. Introduction of these complexes to the capture antibody-immobilized sensor surface resulted in a classic AuNP-based sandwich immunoassay format that has been used for signal amplification. In order to achieve further signal enhancement, a gold staining method was performed, which demonstrated that it is possible to obtain gold staining-mediated signal augmentation on a mass-sensitive device. The sensor response due to gold staining varied as a function of cardiac marker concentration. We also investigated effects of increasing operating frequency on sensor responses. Results showed that detection limit of the SAW sensor could be further improved by increasing the operating frequency.

Continual collection and re-separation of circulating tumor cells from blood using multi-stage multi-orifice flow fractionation

Circulating tumor cells (CTCs) are highly correlated with the invasive behavior of cancer; as such, the ability to isolate and quantify CTCs is of great biomedical importance. This research presents a multi-stage multi-orifice flow fractionation (MS-MOFF) device formed by combining three single-stage multi-orifice segments designed for separating breast cancer cells from blood. The structure and dimensions of the MS-MOFF were determined by hydrodynamic principles to have consistent Reynolds numbers (Re) at each multi-orifice segment. From this device, we achieved improved separation efficiency by collecting and re-separating non-selected target cells in comparison with the single-stage multi-orifice flow fractionation (SS-MOFF). The recovery of breast cancer cells increased from 88.8% to greater than 98.9% through the multi-stage multi-orifice segments. This device can be utilized to isolate rare cells from human blood, such as CTCs, in a label-free manner solely through the use of hydrodynamic forces.

Continuous labeling of circulating tumor cells with microbeads using a vortex micromixer for highly selective isolation.

Circulating tumor cells (CTCs) are identified in transit within the blood stream of cancer patients and have been proven to be a main cause of metastatic disease. Current approaches for the size-based isolation of CTCs have encountered technical challenges as some of the CTCs have a size similar to that of leukocytes and therefore CTCs are often lost in the process. Here, we propose a novel strategy where most of the CTCs are coated by a large number of microbeads to amplify their size to enable complete discrimination from leukocytes. In addition, all of the microbead labeling processes are carried out in a continuous manner to prevent any loss of CTCs during the isolation process. Thus, a microfluidic mixer was employed to facilitate the efficient and selective labeling of CTCs from peripheral blood samples. By generating secondary vortex flows called Taylor-Gortler vortices perpendicular to the main flow direction in our microfluidic device, CTCs were continuously and successfully coated with anti-epithelial cell adhesion molecule-conjugated beads. After the continuous labeling, the enlarged CTCs were perfectly trapped in a micro-filter whereas all of the leukocytes escaped.

Continuous microfluidic airborne bacteria separation using dielectrophoresis

Airborne microbes such as bacteria are a threat to public health. To prevent and control such dangerous biological particles, robust and real-time detection systems are necessary. For direct and real-time detection of airborne microbes, samples must be collected and typically re-suspended in liquid prior to detection; however, environmental particles such as dust are also trapped in such samples. Therefore, the isolation of target bacteria or selective collection of microbes from unwanted non-biological particles prior to detection is of great importance. Dielectrophoresis (DEP), the translational motion of particles in non-uniform electric fields, is an emerging technique that can rapidly separate biological particles in microfluidics. In this paper, we propose a new method for the separation of airborne microbes using DEP with a simple and novel curved electrode design for separating bacteria in a solution containing beads or dust which is taken from an airborne environmental sample. As there has been little research on analyzing environmental samples using microfluidics and DEP, this work describes a novel strategy for a rapid and direct bioaerosol monitoring system.

SAW immunosensors for HBsAb detection

This paper presents the implementation of a surface acoustic wave (SAW) immunosensor system for real sample detection of antibody to hepatitis B surface antigen (HBsAg). The SAW sensor device was based on the mass detection using LOVE wave with the central frequency of 200 MHz. The thickness of the SiO2 guiding layer was optimized both theoretically and experimentally to obtain the maximum sensitivity. A detection circuitry using a differential scheme measured the frequency difference by subtracting the resonant frequency of the sample SAW from that of the reference SAW. The sensitivity of the sensor was 0.74 Hz/(pg/mulscr) and the detection limit was 10 pg/mulscr. Our SAW sensor successfully detected natural anti-HBsAg antidbody (HBsAb) of the real whole blood samples, which opens up a possibility of label-free protein detection without any pretreatment procedures.