Multiplexed Photometry And Fluorimetry Using Multiple Frequency Channels

ABSTRACT MULTIPLEXED PHOTOMETRY AND FLUORIMETRY USING MULTIPLE FREQUENCY CHANNELS by KHALED M. DADESH August 2013 Advisor: Dr. Amar Basu Major: Electrical Engineering Degree: Doctor of Philosophy Multispectral photometry and fluorimetry are useful for quantifying and distinguishing samples during flow injection analysis, flow cytometry, and ratiometric absorbance measurement. However, multispectral detectors, including spectrometers, typically require arrayed or multiple light detectors, optical components, and path alignments, all of which increases the size and cost of the detection system. Several previous efforts have attempted the use of time division multiplexing or frequency division multiplexing (FDM) techniques to minimize both size and cost of multispectral photometry equipment by using only a single light detector. Although many of these designs achieved low cost, they generally operated at \u3c50 KHz, which limited the detection speed of the overall system. An alternative frequency multiplexing design operated at 3MHz; however, it required electro optical modulators [50], which are too expensive and bulky for portable applications. In contrast to both approaches, the objective of this research is to use frequency division multiplexing to perform multispectral photometry and fluorimetry while achieving both low cost and high frequency operation (up to 100 MHz). The multiplexing is performed electronically using low cost optoelectronic sources, a single light detector, and a single high-throughput interrogation window. It enables us to perform multi-parameter biological analysis at lower costs and less complexity. Multiple monochromatic light sources, each with a unique wavelength, are electronically modulated at distinct frequencies, and their combined light emission is directed to the sample detection cell. The light transmitted by the sample (absorbance mode) or emitted by the sample (fluorescence mode) is directed to a single light detector. The received light is then converted to a voltage signal and demodulated into the frequency channels using phase-sensitive electronics. Each recovered channel therefore provides either absorbance or fluorescence at its respective optical wavelength. The system is designed to operate at high speed in order to be used in high throughput detectors such as flow cytometers. As a proof of concept, we apply the FDM technique in two detection systems: 1) a three-color absorbance photometry detector and 2) a two-color laser induced fluorescence (LIF) detector. In the first system, three LEDs are operated with 150 KHz, 200 KHz, and 250 KHz modulation frequencies, and the system achieves a 1 ms measurement time constant at an overall component cost \u3c$10. We perform absorbance photometry of four different organic dyes in flow injected solutions and in discrete droplet microreactors with throughputs in the 10\u27s of samples per second. In both cases, the system is able to simultaneously discriminate between them [13]. In the LIF system, first two laser diodes operated at 1 KHz and 1.5 KHz, respectively, are used to excite fluorophores at the respective frequencies. This system is able to distinguish low speed (1 drop/sec) water-in-oil droplets containing fluorescein or rhodamine-6G generated in a microfluidic junction. Second two laser diodes operated at 25MHz and 40 MHz, respectively, are controlled using a developed high frequency FDM system to excite Fluorescein and Alexa 680 dyes at the respective frequencies. Because of the high frequency operation, this system is able to distinguish alternating high speed (300 drops/sec) droplets containing the two fluorescent dyes. In both case, the developed In previous experiments we use an inverted fluorescence microscope with a specific optical cube to excite dyes and collect fluorescence signals. These two FDM-LIF systems identify the different fluorophores based on their excitation frequency rather than their emission band, giving it a unique ability to distinguish fluorophores with overlapping emission spectra. However, overlapping excitation spectra is a problem in the FDM-LIF system, and any assay has to be prepared using fluorophores with minimal excitation overlap. Therefore, fluorophores with sharp excitation lines such as lanthanide ions are the best candidate material in use with FDM-LIF system. The system uses high frequency (100 MHz) modulation which enables multiplexed time constants on the order of 1 µs. Achieving this high bandwidth allow us to apply the system towards high throughput analysis such as cell cytometry, where it could substantially reduce cost and size of the system. Therefore, the FDM-LIF system is installed in an old BD bioscience cytometer, which is available in the cell cytometry laboratory in Karmanos Cancer Research Center (KCRC) located at Detroit Medical Center (DMC). A biological assay containing Alexa Fluor® 680 Goat Anti-Mouse IgG (H+L) and Alexa Fluor® 430 Goat Anti-Mouse IgG (H+L) with BDTM CompBead Anti-mouse Ig, κ beads is tested using the FDM-LIF system. The system is capable to count the two different antigens simultaneously, which gives the possibility of incorporating this system in cytometers. This technology promises to reduce cost and complexity of future cytometers

Screening methodologies for the determination of water contaminant residues by compact disk technology

[EN] The contamination of water resources with many industrial, agricultural and other chemicals is one of the key environmental problems that humanity is facing nowadays. Despite the fact that they are usually present at very low concentration, they possess a significant risk to aquatic and human life. To address this issue many national and international institutions set different regulations to monitor and control the water quality. Currently, the monitoring of compounds included in official watch lists is conducted by chromatographic and mass spectroscopic methods. These techniques are approved as "gold standards" for analytical quantitation of organic residues in water. Although they are sensitive and reproducible, cannot be used on-site. The need of sampling and centralized laboratory measurements makes not only the overall cost high but lowering the efficiency of the analysis. Therefore, there is an urgent need to develop suitable field methods to facilitate the in situ measurements at a low cost. Biosensors are therefore an alternative technology that can provide sensitive results in a fast and affordable way. This thesis has focused on the development of a biosensor based on immunoassays and compact disk technology, for the multiplex detection of priority water contaminants. As the methods based on the immunorecognition events are challenging in terms of the selectivity and sensitivity, the major part of the thesis was the selection of the immunoreagents, assay form and procedure. For the detection part, gold nanostructures were selected as sensitive tags for signal enhancement. Therefore, different nanoparticles were studied in order to select the optimal size in terms of the signal enhancement, sensitivity and antibody amount used. Also, the assays performances with signal enhancement and without any amplification were evaluated. The best immunoassay was selected for developing the multiplexed assay. Furthermore, an approach to improve the readout sensitivity of microimmunoassays based on used of gold nanoparticles as both capture and detection species was demonstrated. The method is based on the performance of the immunorecognition event in a homogeneous mode and detection part in the heterogeneous format. Finally, representative water samples were analysed to confirm the applicability of the multi-residue assay. The analytical properties have been established for each methodology and the obtained results have been validated by comparison with reference techniques. The investigations carried out in this work, have resulted in new insights in immunoassay technique that could be useful for screening applications.[ES] La contaminación de aguas superficiales causada por plaguicidas y productos industriales es actualmente, uno de los grandes problemas medioambientales. Aunque estas sustancias están presentes a niveles muy bajos, tienen efectos perjudiciales en el medio en general y especialmente en humanos. Por este motivo, diferentes instituciones han regulado los niveles de contaminantes en áreas de controlar de la calidad de las aguas, creando listas prioritarias de sustancias peligrosas y tóxicas para el medio ambiente. Actualmente, la monitorización de los contaminantes incluidos en las listas oficiales se realiza mediante técnicas cromatográficas y espectrometría. Estos métodos analíticos están aprobados como técnicas de referencia para la determinación de residuos orgánicos presentes en aguas naturales. A pesar de ser técnicas fiables, reproducibles y sensibles, los métodos cromatográficos no están exentos de inconvenientes. Este tipo de metodologías requiere una instrumentación costosa y una laboriosa preparación de muestra, que hacen que el análisis sea en general complejo. Por ello, el desarrollo de métodos analíticos alternativos que faciliten hacer medidas in-situ a bajo coste y con gran capacidad de trabajo es de gran utilidad. En este sentido, las técnicas inmunoquímicas tienen un gran potencial analítico ya que son en general sensibles y selectivas, se pueden utilizar en el lugar de la toma de muestra y tienen capacidad multianalito. Esta tesis se ha centrado en el desarrollo de un sistema biosensor, basado en la tecnología de disco compacto, para la detección multianalito de diversos contaminantes prioritarios en aguas naturales. Las limitaciones más críticas para el desarrollo de un biosensor multianalito mediante métodos inmunoquímicos son los relacionados con su sensibilidad y selectividad. Por lo tanto, una parte importante de la tesis se ha centrado en la selección de inmunoreactivos, formato y optimización de diferentes parámetros claves del ensayo. Una estrategia utilizada para aumentar la sensibilidad de los ensayos ha consistido en marcar la inmunoreacción con nanopartículas de oro. Para ello, se ha estudiado diferentes tipos (esféricas y cilíndricas) de distinto tamaño y se han comparado sus prestaciones analíticas (relación señal ruido, sensibilidad etc.) También, se han desarrollado inmunoensayos cuantitativos sin necesidad de amplificación de la señal. Por otro lado, se ha desarrollado una aproximación que hemos denominado "inmunocaptura" basada en el uso de nanopartículas de oro como especie de captura de analitos en disolución y que actúa como marcador de la inmunointeracción que tiene lugar en la fase sólida. Finalmente, se han analizado muestras de agua naturales dopadas con distintos niveles de los analitos objeto de estudio para evaluar la utilidad de las metodologías desarrolladas como herramienta de screening masivo en el área medioambiental. Los resultados obtenidos han sido comparados con los obtenidos mediante las técnicas de referencia. Las investigaciones realizadas han permitido desarrollar nuevos formatos de ensayo y conocimientos inmunoquímicos aplicados a la tecnología de disco compacto, aportando nuevas herramientas de screening que permiten la determinación simultánea de contaminantes en aguas naturales por debajo de las concentraciones establecidas en la normativa europea de calidad de agua.[CA] La contaminació d'aigües superficials causada principalment per plaguicides i altres productes industrials és un dels grans problemes mediambientals actuals. Malgrat que aquestes substàncies estan presents en nivells molt baixos, tenen efectes perjudicials en humans i animals. Per aquest motiu, diferents institucions estatals han regulat els nivells de contaminants en àrees de control de la qualitat de l'aigua, creant llistes prioritàries de substàncies perilloses i tòxiques per al medi ambient. Actualment, la monitorització dels contaminants inclosos en les llistes oficials es realitza mitjançant tècniques cromatogràfiques i d'espectroscòpia de masses. Aquests mètodes analítics estan aprovats com a tècniques de referència per a la determinació de residus orgànics presents en aigües naturals. Malgrat ser tècniques fiables, reproduïbles i sensibles, els mètodes cromatogràfics no estan exempts d'inconvenients. Aquest tipus de metodologies requereix una instrumentació costosa i una laboriosa preparació de mostres que fan que l'anàlisi sigui, en general, complex. Per això, el desenvolupament de mètodes analítics alternatius que facilitin la possibilitat de fer mesures in-situ a baix cost i amb gran capacitat analítica és de gran utilitat. En aquest sentit, les tècniques inmunoquímiques tenen un gran potencial analític ja que són, en general, sensibles i selectives, es poden utilitzar en el lloc de presa de la mostra i tenen capacitat multianalit. Aquesta tesi s'ha centrat en el desenvolupament d'un sistema biosensor, basat en la tecnologia de disc compacte, per a la detecció multianalit de diversos contaminants prioritaris en aigües naturals. Les limitacions més crítiques per al desenvolupament d'un biosensor multianalit mitjançant mètodes inmunoquímics són sensibilitat i selectivitat. Per tant, una part important de la tesi es va centrar en la selecció de inmunoreactius, format i optimització de diferents paràmetres clau de l'assaig. La detecció es va dur a terme mitjançant l'ús de nanopartícules d'or com a marcadors de la inmunointeracció i amplificació de la senyal analítica. S'han estudiat diferents estructures d'or (esferes i cilindres) de diferents tamanys, i s'han comparat les seves prestacions analítiques (relació senyal-soroll, sensibilitat, etc.). També s'han desenvolupat immunoassaigs quantitatius sense necessitat d'amplificació del senyal. Per altra banda, s'ha desenvolupat una aproximació que hem denominat "inmunocaptura", basada en l'ús de nanopartícules d'or com a espècie de captura d'analits en dissolució i que actua com a marcador de la inmunointeracció que té lloc en la fase sòlida. Finalment, s'han analitzat mostres d'aigües naturals dopades amb diferents nivells dels analits objecte d'estudi per avaluar la utilitat de les metodologies desenvolupades com a eina de "screening" massiu en l'àrea mediambiental. Els resultats obtinguts han sigut avaluats per comparació amb els obtinguts mitjançant tècniques de referència. Les investigacions realitzades han permès desenvolupar nous formats d'assaig i coneixements inmunoquímics aplicats a la tecnologia de disc compacte, aportant noves eines de "screening" que permetin la determinació de contaminants en aigües naturals per sota dels límits de concentració establerts per les normes internacionals de la qualitat de l'aigua.Dobosz, PD. (2017). Screening methodologies for the determination of water contaminant residues by compact disk technology [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/79548TESI

Advances in Engineering and Application of Optogenetic Indicators for Neuroscience

Our ability to investigate the brain is limited by available technologies that can record biological processes in vivo with suitable spatiotemporal resolution. Advances in optogenetics now enable optical recording and perturbation of central physiological processes within the intact brains of model organisms. By monitoring key signaling molecules noninvasively, we can better appreciate how information is processed and integrated within intact circuits. In this review, we describe recent efforts engineering genetically-encoded fluorescence indicators to monitor neuronal activity. We summarize recent advances of sensors for calcium, potassium, voltage, and select neurotransmitters, focusing on their molecular design, properties, and current limitations. We also highlight impressive applications of these sensors in neuroscience research. We adopt the view that advances in sensor engineering will yield enduring insights on systems neuroscience. Neuroscientists are eager to adopt suitable tools for imaging neural activity in vivo, making this a golden age for engineering optogenetic indicators. Keywords: optogenetic tools; neuroscience; calcium sensor; voltage sensor; neurotransmitter

Defect and thickness inspection system for cast thin films using machine vision and full-field transmission densitometry

Quick mass production of homogeneous thin film material is required in paper, plastic, fabric, and thin film industries. Due to the high feed rates and small thicknesses, machine vision and other nondestructive evaluation techniques are used to ensure consistent, defect-free material by continuously assessing post-production quality. One of the fastest growing inspection areas is for 0.5-500 micrometer thick thin films, which are used for semiconductor wafers, amorphous photovoltaics, optical films, plastics, and organic and inorganic membranes. As a demonstration application, a prototype roll-feed imaging system has been designed to inspect high-temperature polymer electrolyte membrane (PEM), used for fuel cells, after being die cast onto a moving transparent substrate. The inspection system continuously detects thin film defects and classifies them with a neural network into categories of holes, bubbles, thinning, and gels, with a 1.2% false alarm rate, 7.1% escape rate, and classification accuracy of 96.1%. In slot die casting processes, defect types are indicative of a misbalance in the mass flow rate and web speed; so, based on the classified defects, the inspection system informs the operator of corrective adjustments to these manufacturing parameters. Thickness uniformity is also critical to membrane functionality, so a real-time, full-field transmission densitometer has been created to measure the bi-directional thickness profile of the semi-transparent PEM between 25-400 micrometers. The local thickness of the 75 mm x 100 mm imaged area is determined by converting the optical density of the sample to thickness with the Beer-Lambert law. The PEM extinction coefficient is determined to be 1.4 D/mm and the average thickness error is found to be 4.7%. Finally, the defect inspection and thickness profilometry systems are compiled into a specially-designed graphical user interface for intuitive real-time operation and visualization.M.S.Committee Chair: Tequila Harris; Committee Member: Levent Degertekin; Committee Member: Wayne Dale

A new electronic device for measuring pulse and oxygen concentration

Sensor devices such as pulse oximetry are practical tools used by most healthcare professionals, and even patients. An electronic sensor device that has the capability to measure physiological signs of saturation of arterial oxygen (SpO2) and heart beat rate of the human body has been developed in this study.The hardware design of the sensor device consists of a microcontroller PIC18F452, an external flash memory, and a transceiver unit. The most suitable sensors of red and infra red LEDs are installed on the arms of the plastic clip and functioned with the right resistor values. The red and infra red lights are detected by the photo diode and converted to digital numbers by the Assembly Language software program embedded into the microcontroller PIC18LF452. Those digital numbers are converted to SpO2 value in percentage level. A low power dual operational amplifier LM358 is used to amplify the current signal of the two lights, which depends on the intensity and visibility of the two lights. The output signals are displayed in 16 characters and 2 lines on Hitachi HD44870 compatible liquid crystal display (LCD). In order to display this data on personal computer (PC) monitor, the data is also transmitted via Universal Synchronous Asynchronous Receiver Transmitter (USART) ports of microcontroller to a PC. A Visual Basic 6 programming language is installed in the PC to display the wave forms, the percentage of the SpO2 level, and the pulse rate.Pulse oximetry has a promising future in the healthcare industry. This research enables a more efficient and economical means for managing the healthcare of the growing population

In situ sensors for measurements in the global trosposphere

Current techniques available for the in situ measurement of ambient trace gas species, particulate composition, and particulate size distribution are reviewed. The operational specifications of the various techniques are described. Most of the techniques described are those that have been used in airborne applications or show promise of being adaptable to airborne applications. Some of the instruments described are specialty items that are not commercially-available. In situ measurement techniques for several meteorological parameters important in the study of the distribution and transport of ambient air pollutants are discussed. Some remote measurement techniques for meteorological parameters are also discussed. State-of-the-art measurement capabilities are compared with a list of capabilities and specifications desired by NASA for ambient measurements in the global troposphere

Time-resolved quantitative assays and imaging of enzymes and enzyme substrates using a new europium fluorescent probe for hydrogen peroxide

The thesis describes the development of a novel reversible H2O2 fluorescent probe comprising the ternary complex of europium(III), tetracycline and hydrogen peroxide; and its application in the assays of H2O2, of H2O2 producing oxidases and their substrates, and of H2O2 consuming catalase and its inhibitors. The probe is applied in steady-state intensity-based, time-resolved �gated�, or lifetime-based detection modes both for microplate fluorescence measurement and imaging. The fluorescent probe�s advantages include the reversibility of the EuTc-HP system, the possibility of a kinetic real-time detection of the production and the consumption of H2O2, and the system works best at pH 6.9 - 7.0. It also exhibits the typical spectral characteristics of a ligand-to-europium energy transfer system which include a Stokes shift of ~210 nm, line-like emission, excitation at 380-420 nm (e.g. by the 405-nm blue diode laser), and a µs decay time (~60 µs) facilitating time-resolved fluorometry and imaging. For the probe study, Chapter 1 gives a overview of the state of art of the H2O2 measurements. In the following first part of Chapter 2 is presented the characterization of the fluorescent EuTc-HP probe for its absorbance, circular dichroism and fluorescence spectra, fluorescence lifetime and decay profile, optimal pH and stability, and the influence of temperature, buffers, quenchers and interferents. In the last part of Chapter 6, the peculiar molar ratio of the EuTc-HP probe is further investigated for its possible structure and a com-binatorial approach for discovery of new lanthanide probes is preliminarily proceeded as well. Different assays have been developed for H2O2, glucose, glucose oxidase and catalase, as examples for the detection of enzyme substrates and enzymes. Furthermore, different fluorometric schemes have been applied, such as the steady-state intensity-based detection (Chapter 5, catalase), the time-resolved gated detections (Chapter 3, glucose), the rapid lifetime determination method (Chapter 2, H2O2, novel on microplate) and the time-correlated single photon counting method of the lifetime-based detection (Chapter 2, H2O2), for both microplates and cuvettes, compatible with high-throughput screening. The µs range lifetime of the EuTc-HP probe has greatly facilitated fluorescence imaging, a means for visualization and mapping of the analyte with multiple chemical information. Four schemes of imaging, those are the conventional fluorescence intensity imaging (FII), the time-resolved ("gated") imaging (TRI), the phase delay ratioing imaging (PDI), and the rapid lifetime determination imaging (RLI), have been tested for the quantitative analysis. Hydrogen peroxide, glucose, and glucose oxidase have been determined by the fluorescent imaging system, with Chapter 4 highlighting the fluorescence imaging of glucose oxidase. There are possible further applications in perspective for the EuTc-HP probe. Chapter 6 summarizes the initial attempts, such as glucose oxidase based ELISA, the coupled catalase/glucose oxidase system as a platform for screening, and the construction of microplate arrays and sensors

SIMULATIONS-GUIDED DESIGN OF PROCESS ANALYTICAL SENSOR USING MOLECULAR FACTOR COMPUTING

Many areas of science now generate huge volumes of data that present visualization, modeling, and interpretation challenges. Methods for effectively representing the original data in a reduced coordinate space are therefore receiving much attention. The purpose of this research is to test the hypothesis that molecular computing of vectors for transformation matrices enables spectra to be represented in any arbitrary coordinate system. New coordinate systems are selected to reduce the dimensionality of the spectral hyperspace and simplify the mechanical/electrical/computational construction of a spectrometer. A novel integrated sensing and processing system, termed Molecular Factor Computing (MFC) based near infrared (NIR) spectrometer, is proposed in this dissertation. In an MFC -based NIR spectrometer, spectral features are encoded by the transmission spectrum of MFC filters which effectively compute the calibration function or the discriminant functions by weighing the signals received from a broad wavelength band. Compared with the conventional spectrometers, the novel NIR analyzer proposed in this work is orders of magnitude faster and more rugged than traditional spectroscopy instruments without sacrificing the accuracy that makes it an ideal analytical tool for process analysis. Two different MFC filter-generating algorithms are developed and tested for searching a near-infrared spectral library to select molecular filters for MFC-based spectroscopy. One using genetic algorithms coupled with predictive modeling methods to select MFC filters from a spectral library for quantitative prediction is firstly described. The second filter-generating algorithm designed to select MFC filters for qualitative classification purpose is then presented. The concept of molecular factor computing (MFC)-based predictive spectroscopy is demonstrated with quantitative analysis of ethanol-in-water mixtures in a MFC-based prototype instrument

Microparticle-Based Biosensors for Anthropogenic Analytes

Anthropogenic pollution of water resources and the environment by various hazardous compounds and classes of substances raises concerns about public health impacts and environmental damage. Commercially available, portable and easy-to-use devices to detect and quantify these compounds are rather sparse, but would contribute to comprehensive monitoring and reliable risk assessment. The Soft Colloidal Probe (SCP) assay is a promising platform for the development of portable analytical devices and thus has a great potential for a transfer to industry. This assay is based on the differential deformation of an elastic particle, i.e., the SCP, as a function of analyte concentration, which affects the extent of interfacial interactions between the SCP and a biochip surface. The objective of this work was to adapt this assay for the detection of anthropogenic pollutants. Biomimetic molecular recognition approaches were used based on naturally occurring target proteins that specifically bind the anthropogenic pollutants of interest. This adaptation included the elaboration of strategies for site-specific immobilization of the respective proteins and functionalization of SCPs. In this work, it is demonstrated that the SCP method can be employed for the highly specific and sensitive detection of the critically discussed pesticide glyphosate by using the target enzyme 5-enolpyruvylshikimate-3-phosphate synthase. Furthermore, a specific detection scheme for estrogens and compounds with estrogenic and antiestrogenic activity was developed by harnessing estrogen sulfotransferase as the biomimetic recognition element. In the second part of the thesis, improvements of the SCP sensing methodology are described. These improvements were achieved by accelerating data analysis and developing a novel synthesis method for SCPs that ensures monodisperse particles with superior reproducibility. Rapid extraction of interaction energies is achieved by using a pattern matching algorithm that reduces the time required for data analysis to a fraction. The microfluidics-assisted synthesis of SCPs enables the production of highly monodisperse SCPs with adjustable size and mechanical properties. Various functionalization approaches have been developed that allow easy and modular introduction of functional groups and biomolecules for SCP-based sensing approaches

LAB-ON-A-DISCS FOR QUANTIFICATION OF MICROALGAL LIPIDS AND NATURAL ANTIOXIDANTS OF BEVERAGE SAMPLES

Department of Chemical EngineeringSince the first introduction in 1960s, lab-on-a-disc platform has gained much attention due to its great advantages such as simple operation, rapid reaction, low cost, full integration and automation. Lab-on-a-disc has been applied for various research fields such as biomedical, environmental, energy and food agricultural field. In this thesis, fully integrated lab-on-a-disc platform was demonstrated for quantification of microalgal lipid and natural antioxidants from beverage samples, and disc analyzer was introduced to operate the on-disc optical detection. For microalgae lipid quantification, fully automated lab-on-a-disc platform was developed for rapid on-site quantification of lipids from microalgal samples. The whole serial process for microalgal lipid quantification was integrated on lab-on-a-disc platform. Lab-on-a-disc operation time was 13 min. To integrate liquid-solvent extraction of microalgal lipid on a disc, the organic solvent compatible (for ethanol and n-hexane) lab-on-a-disc was newly fabricated. Fabrication technique was developed by combining thermal fusion bonding and laser printed carbon dot based valving. For natural antioxidant determination from beverage samples, lab-on-a-disc platform was developed to integrate the complicated determination steps of antioxidant activity (AA) and the total phenolic content (TPC) from beverage sample. Different beverage samples including wine, beer, various fruit juices and tea were analyzed using our lab-on-a-disc platform. For disc analyzer, on-disc optical signal detecting instrument was developed. It can detect on-disc signal of absorbance and chemiluminescence from result solution. The trials and errors to develop the disc analyzer for following researchers was demonstrated. From both lab-on-a-disc quantification methods, it is proved that lab-on-a-disc can be effectively used for quantification of microalgal biofuel and food antioxidants. It is expected that our lab-on-a-disc based microalgal lipid quantification works give considerable contribution to the commercial production of microalgal-based biofuels by providing rapid, cost-effective, user-friendly on-site quantification of microalgal lipids. Also, our on-disc natural antioxidant detection works would also contribute to functional beverage industry as providing fast, simple determination of natural antioxidants from beverage sample. Both lab-on-a-disc was fabricated by the organic solvent compatible (for ethanol and n-hexane) manner, and it can be used for chemical engineering fields for quality control.clos