Jong-ryul Choi

Fluorescence optical detection in situ for real-time monitoring of cytochrome P450 enzymatic activity of liver cells in multiple microfluidic devices

We describe an in situ fluorescence optical detection system to demonstrate real‐time and non‐invasive detection of reaction products in a microfluidic device while under perfusion within a standard incubator. The detection system is designed to be compact and robust for operation inside a mammalian cell culture incubator for quantitative detection of fluorescent signal from microfluidic devices. When compared to a standard plate reader, both systems showed similar biphasic response curves with two linear regions. Such a detection system allows real‐time measurements in microfluidic devices with cells without perturbing the culture environment. In a proof‐of‐concept experiment, the cytochrome P450 1A1/1A2 activity of a hepatoma cell line (HepG2/C3A) was monitored by measuring the enzymatic conversion of ethoxyresorufin to resorufin. The hepatoma cell line was embedded in MatrigelTM construct and cultured in a microfluidic device with medium perfusion. The response of the cells, in terms of P450 1A1/1A2 activity, was significantly different in a plate well system and the microfluidic device. Uninduced cells showed almost no activity in the plate assay, while uninduced cells in MatrigelTM with perfusion in a microfluidic device showed high activity. Cells in the plate assay showed a significant response to induction with 3‐Methylcholanthrene while cells in the microfluidic device did not respond to the inducer. These results demonstrate that the system is a potentially useful method to measure cell response in a microfluidic system. Biotechnol. Bioeng. 2009; 104: 516–525

Microfluidic assay-based optical measurement techniques for cell analysis: A review of recent progress.

Since the early 2000s, microfluidic cell culture systems have attracted significant attention as a promising alternative to conventional cell culture methods and the importance of designing an efficient detection system to analyze cell behavior on a chip in real time is raised. For this reason, various measurement techniques for microfluidic devices have been developed with the development of microfluidic assays for high-throughput screening and mimicking of in vivo conditions. In this review, we discuss optical measurement techniques for microfluidic assays. First of all, the recent development of fluorescence- and absorbance-based optical measurement systems is described. Next, advanced optical detection systems are introduced with respect to three emphases: 1) optimization for long-term, real-time, and in situ measurements; 2) performance improvements; and 3) multimodal analysis conjugations. Moreover, we explore presents future prospects for the establishment of optical detection systems following the development of complex, multi-dimensional microfluidic cell culture assays to mimic in vivo tissue, organ, and human systems.

Slewing Mirror Telescope optics for the early observation of UV/optical photons from Gamma-Ray Bursts

We report on design, manufacture, and testing of a Slewing Mirror Telescope (SMT), the first of its kind and a part of Ultra-Fast Flash Observatory-pathfinder (UFFO-p) for space-based prompt measurement of early UV/optical light curves from Gamma-Ray Bursts (GRBs). Using a fast slewing mirror of 150 mm diameter mounted on a 2 axis gimbal stage, SMT can deliver the images of GRB optical counterparts to the intensified CCD detector within 1.5~1.8 s over ± 35 degrees in the slewing field of view. Its Ritchey-Chrétien telescope of 100 mm diameter provides a 17 × 17 arcmin² instantaneous field of view. Technical details of design, construction, the laboratory performance tests in space environments for this unique SMT are described in conjunction with the plan for in-orbit operation onboard the Lomonosov satellite in 2013.

Surface plasmon-enhanced nanoscopy of intracellular cytoskeletal actin filaments using random nanodot arrays

The feasibility of super-resolution microscopy has been investigated based on random localization of surface plasmon using blocked random nanodot arrays. The resolution is mainly determined by the size of localized fields in the range of 100-150 nm. The concept was validated by imaging FITC-conjugated phalloidin that binds to cellular actin filaments. The experimental results confirm improved resolution in reconstructed images. Effect of far-field registration on image reconstruction was also analyzed. Correlation between reconstructed images was maintained to be above 81% after registration. Nanodot arrays are synthesized by temperature-annealing without sophisticated lithography and thus can be mass-produced in an extremely large substrate. The results suggest a super-resolution imaging technique that can be accessible and available in large amounts.

A Localized Surface Plasmon Resonance Sensor Using Double-Metal-Complex Nanostructures and a Review of Recent Approaches

From active developments and applications of various devices to acquire outside and inside information and to operate based on feedback from that information, the sensor market is growing rapidly. In accordance to this trend, the surface plasmon resonance (SPR) sensor, an optical sensor, has been actively developed for high-sensitivity real-time detection. In this study, the fundamentals of SPR sensors and recent approaches for enhancing sensing performance are reported. In the section on the fundamentals of SPR sensors, a brief description of surface plasmon phenomena, SPR, SPR-based sensing applications, and several configuration types of SPR sensors are introduced. In addition, advanced nanotechnology- and nanofabrication-based techniques for improving the sensing performance of SPR sensors are proposed: (1) localized SPR (LSPR) using nanostructures or nanoparticles; (2) long-range SPR (LRSPR); and (3) double-metal-layer SPR sensors for additional performance improvements. Consequently, a high-sensitivity, high-biocompatibility SPR sensor method is suggested. Moreover, we briefly describe issues (miniaturization and communication technology integration) for future SPR sensors.

Investigation of portable in situ fluorescence optical detection for microfluidic 3D cell culture assays

A portable fluorescence optical detection system was developed to demonstrate real-time in situ analysis of cells that are three-dimensionally cultured in an extracellular matrix under microfluidic environment. The system was designed to provide a large field of view in the lateral plane to average out cellular processes in an axial layer and simultaneously diffraction-limited axial resolution. In this proof-of-concept study, the detection system was applied to quantitative analyses of short-term measurements of cell staining and cell cytotoxicity and long-term monitoring of a cell-invasion assay. For assays, colon cancer cells were cultured in a Matrigel or alginate matrix. The measured data were largely consistent with predicted results and revealed quantitatively cell dynamics specific to 3D cell cultures. The detection system has a potential as a single package to investigate 3D cultures in a microfluidic system.

Recent advances of nanostructure implemented spectroscopic sensors—A brief overview

ABSTRACT By the conjugation of requirements of high-performance sensing platforms and developments of nanotechnology, various nanostructure implemented photonic, spectroscopic sensors have been investigated. In this review article, we address 3 types of nanostructure implemented optical sensor techniques—localized surface plasmon resonance (LSPR) sensors, extraordinary optical transmission (EOT) based sensors, and Raman-based spectroscopic sensors—and recent advances of the nanostructure assisted sensors arranged by 2 important issues: the employment of novel nanostructures and the application of newly investigated fabrication techniques for larger sensing area.

Current achievements of nanoparticle applications in developing optical sensing and imaging techniques

Metallic nanostructures have recently been demonstrated to improve the performance of optical sensing and imaging techniques due to their remarkable localization capability of electromagnetic fields. Particularly, the zero-dimensional nanostructure, commonly called a nanoparticle, is a promising component for optical measurement systems due to its attractive features, e.g., ease of fabrication, capability of surface modification and relatively high biocompatibility. This review summarizes the work to date on metallic nanoparticles for optical sensing and imaging applications, starting with the theoretical backgrounds of plasmonic effects in nanoparticles and moving through the applications in Raman spectroscopy and fluorescence biosensors. Various efforts for enhancing the sensitivity, selectivity and biocompatibility are summarized, and the future outlooks for this field are discussed. Convergent studies in optical sensing and imaging have been emerging field for the development of medical applications, including clinical diagnosis and therapeutic applications.

Plasmonic signal enhancements using randomly distributed nanoparticles on a stochastic nanostructure substrate

ABSTRACT The surface-enhanced Raman spectrum was investigated through a numerical model and experiments constructed based on the stochastic Ag nanoislands (AgNIs) substrate. By a rigorous coupled-wave analysis (RCWA) method, the basic properties of electric field were calculated for numerical analysis. The plasmonic coupling between Au nanoparticles (AuNPs) and AgNI substrate was optimized by changing the position of AuNPs on the Ag nanostructured substrate. Furthermore, we experimentally confirmed that AgNIs substrate enable that the intensity of Raman spectra were dramatically improved up to ∼20-fold compared to that of a silver thin film as we expected in numerical calculations. The results gained in this work suggest that we could significantly enhance the Raman signal using easily fabricable AgNI substrates, and can provide the potential applications, such as food, pharmaceutical, and security inspections.

Enhanced image reconstruction of three-dimensional fluorescent assays by subtractive structured-light illumination microscopy.

We investigate improved image reconstruction of structured light illumination for high-resolution imaging of three-dimensional (3D) cell-based assays. For proof of concept, an in situ fluorescence optical detection system was built with a digital micromirror device as a spatial light modulator, for which phase and tilting angle in a grid pattern were varied to implement specific image reconstruction schemes. Subtractive reconstruction algorithms based on structured light illumination were used to acquire images of fluorescent microbeads deposited as a two-dimensional monolayer or in 3D alginate matrix. We have confirmed that an optical subtraction algorithm improves axial and lateral resolution by effectively removing out-of-focus fluorescence. The results suggest that subtractive image reconstruction can be useful for structured illumination microscopy of broad types of cell-based assays with high image resolution.