Handheld Fluorometer Research Articles

Portable Aptasensor Based on Parallel Rolling Circle Amplification for Tumor-Derived Exosomes Liquid Biopsy.

Here, a separation-free and label-free portable aptasensor is developed for rapid and sensitive analysis of tumor-derived exosomes (TEXs). It integrated a parallel rolling circle amplification (RCA) reaction, selective binding of metal ions or small molecules to nucleic acid-specific conformations, and a low-cost, highly sensitive handheld fluorometer. Lung cancer, for example, is targeted with two typical biomarkers (mucin 1 and programmed cell death ligand 1 (PD-L1)) on its exosomes. The affinity of aptamers to the targets modulated the amount of RCA products (T-Hg2+-T and cytosine (C)-rich single-stranded DNA), which in turn affected the fluorescence intensity of quantum dots (QDs) and methylene blue (MB). The results revealed that the limit of detection (LOD) of the handheld fluorometer for cell-derived exosomes can be as low as 30 particlesmL-1. Moreover, its specificity, sensitivity, and area under the curve (AUC) are 93% (14/15), 92% (23/25), and 0.956, as determined by the analysis of 40 clinical samples. Retesting 16 of these samples with the handheld fluorometer yielded strong concordance between the fluorometer results and those acquired from clinical computed tomography (CT) and pathology.

Single-Cell Liquid Biopsy of Lung Cancer: Ultra-Simplified Efficient Enrichment of Circulating Tumor Cells and Hand-Held Fluorometer Portable Testing.

Herein, we propose a paper-based laboratory via enzyme-free nucleic acid amplification and nanomaterial-assisted cation exchange reactions (CERs) assisted single-cell-level analysis (PLACS). This method allowed for the rapid detection of mucin 1 and trace circulating tumor cells (CTCs) in the peripheral blood of lung cancer patients. Initially, an independently developed method requiring one centrifuge, two reagents (lymphocyte separation solution and erythrocyte lysate), and a three-step, 45 min sample pretreatment was employed. The core of the detection approach consisted of two competitive selective identifications: copper sulfide nanoparticles (CuS NPs) to C-Ag+-C and Ag+, and dual quantum dots (QDs) to Cu2+ and CuS NPs. To facilitate multimodal point-of-care testing (POCT), we integrated solution visualization, test strip length reading, and a self-developed hand-held fluorometer readout. These methods were detectable down to ag/mL of mucin 1 concentration and the single-cell level. Forty-seven clinical samples were assayed by fluorometer, yielding 94% (30/32) sensitivity and 100% (15/15) specificity with an area under the curve (AUC) of 0.945. Nine and 15 samples were retested by a test strip and hand-held fluorometer, respectively, with an AUC of 0.95. All test results were consistent with the clinical imaging and the folate receptor (FR)-PCR kit findings, supporting its potential in early diagnosis and postoperative monitoring.

Smartphone and handheld fluorometer enable rapid point of care testing of Escherichia coli in urinary tract infections via specific proteolytic cleavage and cascade amplifications

Culture-free, rapid, and ultrasensitive quantification of Escherichia coli (E. coli) is urgently needed to diagnose clinical urinary tract infections (UTIs), especially point-of-care testing (POCT). Herein, a homogeneous sensitive multi-modal (fluorometer, smartphone, and self-developed handheld fluorometer) assay was developed to detect E. coli within 30 min, and nanoscale luminescent cadmium telluride quantum dots (CdTe QDs) were employed as signal reporters. E. coli outer membrane protease T (OmpT) is a biomarker for the specific proteolytic cleavage of peptides. p-Aminophenol (PAP), generated by leucine aminopeptidase (LAP) via secondary cleavage, was used to reduce Ag+. Cascade amplifications included Ag+ and copper sulfide nanoparticles (CuS NPs) to release Cu2+, CdTe QDs, and Cu2+/CuS NPs, which can ensure detection sensitivity. The method achieved single bacterium detection with great stability and selectivity. The fluorometer, handheld fluorometer and smartphone red-green-blue (RGB) value-based modes were determined to be 50, 15, and 15 clinical urine samples, respectively. The findings were consistent with clinical culture counting, and an area under the curve (AUC) of 0.95 with 90% sensitivity and 100% specificity was achieved. Overall, this OmpT-based catalytic biosensor can provide a solution for the POCT diagnosis of UTIs based on E. coli detection.

Development of a microfluidic device to enrich and detect zearalenone in food using quantum dot-embedded molecularly imprinted polymers.

Mycotoxins are secondary metabolites of certain moulds, prevalent in 60-80% of food crops and many processed products but challenging to eliminate. Consuming mycotoxin-contaminated food and feed can lead to various adverse effects on humans and livestock. Therefore, testing mycotoxin residue levels is critical to ensure food safety. Gold standard analytical methods rely on liquid chromatography coupled with optical detectors or mass spectrometers, which are high-cost with limited capacity. This study reported the successful development of a microfluidic "lab-on-a-chip" device to enrich and detect zearalenone in food samples based on the fluorescence quenching effect of quantum dots and selective affinity of molecularly imprinted polymers (MIPs). The dummy template and functional polymer were synthesized and characterized, and the detailed microfluidic chip design and optimization of the flow conditions in the enrichment module were discussed. The device achieved an enrichment factor of 9.6 (±0.5) in 10 min to quantify zearalenone spiked in food with high recoveries (91-105%) at 1-10 mg kg-1, covering the concerned residue levels in the regulations. Each sample-to-answer test took only 20 min, involving 3 min of manual operation and no advanced equipment. This microfluidic device was mostly reusable, with a replaceable detection module compatible with fluorescence measurement using a handheld fluorometer. To our best knowledge, the reported device was the first application of an MIP-based microfluidic sensor for detecting mycotoxin in real food samples, providing a novel, rapid, portable, and cost-effective tool for monitoring mycotoxin contamination for food safety and security.

Fructose@histone synergistically improve the performance of DNA-templated Cu NPs: rapid analysis of LAM in tuberculosis urine samples using a handheld fluorometer and a smartphone RGB camera.

This study presented a nanoparticle-enhanced aptamer-recognizing homogeneous detection system combined with a portable instrument (NASPI) to quantify lipoarabinomannan (LAM). This system leveraged the high binding affinity of aptamers, the high sensitivity of nanoparticle cascade amplification, and the stabilization effect of dual stabilizers (fructose and histone), and used probe-Cu2+ to achieve LAM detection at concentrations ranging from 10 ag mL-1 to 100 fg mL-1, with a limit of detection of 3 ag mL-1 using a fluorometer. It can also be detected using an independently developed handheld fluorometer or the red-green-blue (RGB) camera of a smartphone, with a minimum detection concentration of 10 ag mL-1. We validated the clinical utility of the biosensor by testing the LAM in the urine of patients. Forty urine samples were tested, with positive LAM results in the urine of 18/20 tuberculosis (TB) cases and negative results in the urine of 6/10 latent tuberculosis infection cases and 10/10 non-TB cases. The assay results revealed a 100% specificity and a 90% sensitivity, with an area under the curve of 0.9. We believe that the NASPI biosensor can be a promising clinical tool with great potential to convert LAM into clinical indicators for TB patients.

Diagnosis of Induced Resistance State in Tomato Using Artificial Neural Network Models Based on Supervised Self-Organizing Maps and Fluorescence Kinetics.

The aim of this study was to develop three supervised self-organizing map (SOM) models for the automatic recognition of a systemic resistance state in plants after application of a resistance inducer. The pathosystem Fusarium oxysporum f. sp. radicis-lycopersici (FORL) + tomato was used. The inorganic, defense inducer, Acibenzolar-S-methyl (benzo-[1,2,3]-thiadiazole-7-carbothioic acid-S-methyl ester, ASM), reported to induce expression of defense genes in tomato, was applied to activate the defense mechanisms in the plant. A handheld fluorometer, FluorPen FP 100-MAX-LM by SCI, was used to assess the fluorescence kinetics response of the induced resistance in tomato plants. To achieve recognition of resistance induction, three models of supervised SOMs, namely SKN, XY-F, and CPANN, were used to classify fluorescence kinetics data, in order to determine the induced resistance condition in tomato plants. To achieve this, a parameterization of fluorescence kinetics curves was developed corresponding to fluorometer variables of the Kautsky Curves. SKN was the best supervised SOM, achieving 97.22% to 100% accuracy. Gene expression data were used to confirm the accuracy of the supervised SOMs.

Performance of a Handheld Chlorophyll-a Fluorometer: Potential Use for Rapid Algae Monitoring

Chlorophyll-a measurements are an important factor in the water quality monitoring of surface waters, especially for determining the trophic status and ecosystem management. However, a collection of field samples for extractive analysis in a laboratory may not fully represent the field conditions. Handheld fluorometers that can measure chlorophyll-a in situ are available, but their performance in waters with a variety of potential light-interfering substances has not yet been tested. We tested a handheld fluorometer for sensitivity to ambient light and turbidity and compared these findings with EPA Method 445.0 using water samples obtained from two urban lakes in Tucson, Arizona, USA. Our results suggested that the probe was not sensitive to ambient light and performed well at low chlorophyll-a concentrations (<25 µg/L) across a range of turbidity levels (50–70 NTU). However, the performance was lower when the chlorophyll-a concentrations were >25 µg/L and turbidity levels were <50 NTU. To account for this discrepancy, we developed a calibration equation to use for this handheld fluorometer when field monitoring for potential harmful algal blooms in water bodies.

Chlorophyll fluorescence imaging captures photochemical efficiency of grain sorghum (Sorghum bicolor) in a field setting

BackgroundPhotosynthesis is one of the most important biological reactions and forms the basis of crop productivity and yield on which a growing global population relies. However, to develop improved plant cultivars that are capable of increased productivity, methods that can accurately and quickly quantify photosynthetic efficiency in large numbers of genotypes under field conditions are needed. Chlorophyll fluorescence imaging is a rapid, non-destructive measurement that can provide insight into the efficiency of the light-dependent reactions of photosynthesis.ResultsTo test and validate a field-deployed fluorescence imaging system on the TERRA-REF field scanalyzer, leaves of potted sorghum plants were treated with a photosystem II inhibitor, DCMU, to reduce photochemical efficiency (FV/FM). The ability of the fluorescence imaging system to detect changes in fluorescence was determined by comparing the image-derived values with a handheld fluorometer. This study demonstrated that the imaging system was able to accurately measure photochemical efficiency (FV/FM) and was highly correlated (r = 0.92) with the handheld fluorometer values. Additionally, the fluorescence imaging system was able to track the decrease in photochemical efficiency due to treatment of DCMU over a 7 day period.ConclusionsThe system’s ability to capture the temporal dynamics of the plants’ response to this induced stress, which has comparable dynamics to abiotic and biotic stressors found in field environments, indicates the system is operating correctly. With the validation of the fluorescence imaging system, physiological and genetic studies can be undertaken that leverage the fluorescence imaging capabilities and throughput of the field scanalyzer.

Highly Portable and Sensitive Filter-Based Antibody-or DNA Aptamer-Enzyme-Linked Fluorescence Detection of Potential Bacterial Pathogens Proximal to Agricultural Fields

A novel, sensitive, rapid and highly portable system consisting of a membrane filtration unit connected to a large syringe with tubing was used to detect Escherchia coli and Salmonella enterica bacteria in a mobile laboratory setting. These simple fluorescent enzyme-linked molecular assays consisted of biotinylated DNA aptamers or antibodies which were allowed to bind the target bacteria and be deposited on 0.45 μm pore size membrane filters by suction, followed by binding of streptavidin-peroxidase conjugate, wash steps and development in Amplex® UltraRed (AUR; resazurin derivative) with hydrogen peroxide activation. Fluorescence values of developed AUR from the filter surface were assessed with a commercially-available handheld fluorometer. Preliminary data indicate detection limits in the range of 50–110 E. coli and Salmonella bacterial cells per sample with good specificity and total processing times under one hour.

Predicting cyanobacterial biovolumes from phycocyanin fluorescence using a handheld fluorometer in the field

Toxic cyanobacterial blooms are becoming more prevalent in freshwater systems, increasing the need for monitoring to protect human health. Phycocyanin fluorescence sensors have been developed as tools for providing fast and cost-effective proxy measurements for cyanobacterial biomass. However, poor precision and low sensitivity in many of the probe sensors assessed to-date has restricted their potential for practical application in cyanobacterial monitoring programmes. In the present study, the sensitivity and accuracy of a handheld fluorometer, the CyanoFluor, was assessed using 12 cyanobacterial strains and samples from four different lakes collected weekly for 12 weeks. After the initial measurements, the samples were lysed by sonication, which we hypothesised would reduce inter and intra-specific differences. The CyanoFluor displayed high sensitivity (limit of quantification = 3.5 µg L−1 of phycocyanin) and was able to detect cyanobacterial biovolumes to levels much lower than the threshold levels in current recreational guidelines worldwide. There were strong and significant phycocyanin to biovolume relationships (r2 ≥ 0.88, P < 0.05) for all 12 cyanobacterial cultures. Collectively, strong relationships between phycocyanin fluorescence and cyanobacterial biovolumes were also identified in environmental samples (r2 ≥ 0.78, P < 0.001), although weaker relationships were identified when lakes were analysed separately (r2 = 0.06 – 0.90). There were differences in phycocyanin per biovolume between both cultured strains and lakes, highlighting innate interspecific differences that exist between cyanobacterial species. Lysis of samples consistently reduced variability between technical replicates, in cyanobacteria cultures (up to 87% reduction in sample variability) and environmental samples (71 – 93% reduction), indicating that it would be a useful methodological step to improve the repeatability of results. When guideline thresholds (aligned with currently enforced risk assessment categories) were modelled based on the most successful linear regression model, 74% of samples were assigned to the correct risk category. The sensitivity of the CyanoFluor and accuracy of the phycocyanin threshold models, indicates high potential for this method to be integrated into cyanobacterial monitoring programmes.

Handheld fluorometer for in-situ melamine detection via interference synthesis of dsDNA-templated copper nanoparticles.

Fluorescent copper nanoparticles templated by dsDNA have gained significant research interest as they are inexpensive and easy to synthesize, and have found applications in the detection of a wide range of analytes. The presence of the analyte in the reaction mixture interferes with the synthesis of the copper nanoparticles and the subsequent drop in fluorescence can be correlated to the concentration of the analyte present in the solution. Analyte detection using copper nanoparticle-based assays is amenable for in-situ applications as the test does not require expensive reagents and can be performed at room temperature. However, expensive and sophisticated detection systems are required for the detection of copper nanoparticles due to the low fluorescence emission signal from these nanoparticles. This restricts the use of the technology to centralized labs. Utilizing a recently developed chemical technique for fluorescence enhancement, this paper presents the first report of a handheld fluorometer capable of detecting DNA-templated copper nanoparticles. The fluorometer is portable and constructed with low-cost, off-the-shelf components like a UV-LED and a PIN photodiode. The performance of the developed system is demonstrated through the detection of melamine in milk samples via the interference synthesis of copper nanoparticles. Melamine is an adulterant used in dairy products that is harmful to human health if present in levels above 1ppm. The developed system is capable of detecting up to 0.1ppm of melamine in milk samples with a linear relationship observed between the detector output and concentration of melamine in the range from 0.1ppm to 100ppm (R2=0.9979).

Establishment and application of a novel fluorescence-based analytical method for the rapid detection of viable bacteria in different samples.

A rapid method for readily detecting the total numbers of viable bacterial cells in numerous samples (including surface water, solid inoculants, and soil samples) is reported using a newly developed hand-held fluorometer and a fluorescent dye Calcein UltraGreen™ AM. Compared to the traditional plate counting method that requires 48 hours of cultivation, the newly established method does not require any incubation time, making the detection method faster and more convenient. The portable rapid detection fluorometer has a wide dynamic range of relative fluorescence intensity from 45 to 30 133. It can detect bacterial concentration ranging from 105 to 1010 cells per mL. This newly established method has good applicability for accurately and quickly detecting the cell number of viable bacteria in various samples. The results of the fluorescence-based method were compared with those of the traditional plate counting method, and it was found that the relative standard deviation was less than 6%. This new rapid measurement system provides a robust method for the rapid on-site detection of viable bacteria.

Development of a fluorescent enzyme-linked DNA aptamer-magnetic bead sandwich assay and portable fluorometer for sensitive and rapid listeria detection.

A fluorescent DNA aptamer-magnetic bead sandwich assay was developed to detect listeriolysin O (LLO) protein from pathogenic Listeria bacteria using a peroxidase-linked system, Amplex Ultra Red (AUR; derivatized resazurin) substrate, and a custom-designed handheld fluorometer. The assay is highly sensitive with demonstrated limits of detection (LODs) in the range of 4 to 61 L. monocytogenes cells or the equivalent LLO produced by 4 to 61 cells on average in separate titration trials. Total assay processing and analysis time was approximately 30 mins. The assay has demonstrated the ability to detect 6 species of Listeria as desired by the USDA's Food Safety Inspection Service (FSIS). The portable system was designed to be used primarily with surface swab samples from fomites, but it can also be used to assess enrichment cultures. The minimal time to detect a positive enrichment culture in our hands from an initial 10 cell inoculum in 200 ml of broth has been 8 h post-incubation at 37 °C in shaker flask cultures. An optional automated magnetic bead assay processing and wash device capable of simultaneously processing 6 samples with low and consistent fluorescence background for higher volume central laboratories is also described.

Molecular diagnostics in a teacup: Non-Instrumented Nucleic Acid Amplification (NINA) for rapid, low cost detection of Salmonella enterica.

We report on the use of a novel non-instrumented platform to enable a Loop Mediated isothermal Amplification (LAMP) based assay for Salmonella enterica. Heat energy is provided by addition of a small amount (<150 g) of boiling water, and the reaction temperature is regulated by storing latent energy at the melting temperature of a lipid-based engineered phase change material. Endpoint classification of the reaction is achieved without opening the reaction tube by observing the fluorescence of sequence-specific FRET-based assimilating probes with a simple handheld fluorometer. At or above 22°C ambient temperature the non-instrumented devices could maintain reactions above a threshold temperature of 61°C for over 90 min-significantly longer than the 60 min reaction time. Using the simple format, detection limits were less than 20 genome copies for reactions run at ambient temperatures ranging from 8 to 36°C. When used with a pre-enrichment step and non-instrumented DNA extraction device, trace contaminations of Salmonella in milk close to 1 CFU/mL could be reliably detected. These findings illustrate that the non- instrumented amplification approach is a simple, viable, low-cost alternative for field-based food and agricultural diagnostics or clinical applications in developing countries.

In-Field Implementation of a Recombinant Factor C Assay for the Detection of Lipopolysaccharide as a Biomarker of Extant Life within Glacial Environments.

The discovery over the past two decades of viable microbial communities within glaciers has promoted interest in the role of glaciers and ice sheets (the cryosphere) as contributors to subglacial erosion, global biodiversity, and in regulating global biogeochemical cycles. In situ or in-field detection and characterisation of microbial communities is becoming recognised as an important approach to improve our understanding of such communities. Within this context we demonstrate, for the first time, the ability to detect Gram-negative bacteria in glacial field-environments (including subglacial environments) via the detection of lipopolysaccharide (LPS); an important component of Gram-negative bacterial cell walls. In-field measurements were performed using the recently commercialised PyroGene® recombinant Factor C (rFC) endotoxin detection system and used in conjunction with a handheld fluorometer to measure the fluorescent endpoint of the assay. Twenty-seven glacial samples were collected from the surface, bed and terminus of a low-biomass Arctic valley glacier (Engabreen, Northern Norway), and were analysed in a field laboratory using the rFC assay. Sixteen of these samples returned positive LPS detection. This work demonstrates that LPS detection via rFC assay is a viable in-field method and is expected to be a useful proxy for microbial cell concentrations in low biomass environments.

小型クロロフィル蛍光測定器を用いた水ストレスによる樹勢衰退の客観的診断の可能性

小型クロロフィル蛍光測定器を用いた水ストレスによる樹勢衰退の客観的診断の可能性

An aptamer beacon responsive to botulinum toxins

Sixty candidate DNA aptamers were developed against botulinum neurotoxin (BoNT) type A light chain (LC) from ten rounds of selection, resulting in several identical sequences. Secondary structures of the identical aptamers were compared to structures of previously reported BoNT A DNA aptamers. A series of ten candidate loop structures were selected from this comparison as potential binding pockets and aptamer beacons. These candidate beacons were synthesized with 5′-TYE 665 and 3′-Iowa Black quencher labels for comparison of fluorescence levels as a function of BoNT A LC concentration. Only three of the ten candidates exhibited any fluorescence response to increasing levels of BoNT A LC. However, of the two most responsive candidates, one represented a subset loop of the larger more intensely fluorescent double-looped structure, designated Beacon 10. This beacon yielded a lower limit of detection of 1ng/mL in buffer using a spectrofluorometer and a portable handheld fluorometer, but also responded substantially to BoNT A, B, E holotoxins and heavy or light chain components even in a dilute soil suspension, but not in 50% human serum. Beacon 10 did not respond strongly to a variety of other divergent peptides, suggesting that it is relatively specific to the level of botulinum toxins and is only useful for environmental testing. Beacon 10 also shared short sequence segments with other published BoNT aptamer DNA sequences, suggesting that these may be points of physical contact between the aptamers and BoNTs.

DNA Aptamer Beacon Assay for C-Telopeptide and Handheld Fluorometer to Monitor Bone Resorption

A novel DNA aptamer beacon is described for quantification of a 26-amino acid C-telopeptide (CTx) of human type I bone collagen. One aptamer sequence and its reverse complement dominated the aptamer pool (31.6% of sequenced clones). Secondary structures of these aptamers were examined for potential binding pockets. Three-dimensional computer models which analyzed docking topologies and binding energies were in agreement with empirical fluorescence experiments used to select one candidate loop for beacon assay development. All loop structures from the aptamer finalists were end-labeled with TYE 665 and Iowa Black quencher for comparison of beacon fluorescence levels as a function of CTx concentration. The optimal beacon, designated CTx 2R-2h yielded a low ng/ml limit of detection using a commercially available handheld fluorometer. The CTx aptamer beacon bound full-length 26-amino acid CTx peptide, but not a shorter 8-amino acid segment of CTx peptide which is a common target for commercial CTx ELISA kits. The prototype assay was shown to detect CTx peptide from human urine after creatinine and urea were removed by size-exclusion chromatography to prevent nonspecific denaturing of the aptamer beacon. This work demonstrates the potential of aptamer beacons to be utilized for rapid and sensitive bone health monitoring in a handheld or point-of-care format.

A hand-held electronic tongue based on fluorometry for taste assessment of tea

A hand-held electronic tongue was developed for determining taste levels of astringency and umami in tea infusions. The sensing principles are based on quenching the fluorescence of 3-aminophthalate by tannin, and the fluorogenic reaction of o-phthalaldehyde (OPA) with amino acids to determine astringency and umami levels, respectively. Both reactions were measured by a single fluorescence sensing system with same excitation and emission wavelengths (340/425 nm). This work describes in detail the design, fabrication, and performance evaluation of a hand-held fluorometer with an ultra-violet light emitted diode (UVLED) and a photo-detector with a filter built-in. The dimension and the weight of proposed electronic tongue prototype are only 120 × 60 × 65 mm 3 and 150 g, respectively. The detection limits of this prototype for theanine and tannic acid were 0.2 μg/ml and 1 μg/ml, respectively. Correlation coefficients of this prototype compared with a commercial fluorescence instrument are both higher than 0.995 in determinations of tannin acid and theanine. Linear detection ranges of the hand-held fluorometer for tannic acid and theanine are 1–20 μg/ml and 0.2–10 μg/ml (CV < 5%, n = 3), respectively. A specified taste indicator for tea, defined as ratio of umami to astringency, was adopted here to effectively distinguish flavour quality of partially fermented Oolong teas.

Application of fluorescent protein-tagged trans factors and immobilized cis elements to monitoring of toxic metals based on in vitro protein–DNA interactions

Environmental toxic metals cause serious global public health problems. On-site monitoring protects people from exposure to such harmful elements. In this study, the bacterial transcriptional switches were applied to monitoring of toxic metals. ArsR and CadC, trans factors of Escherichia coli and Staphylococcus aureus, were fused to GFP. The fusion proteins, ArsR–GFP and CadC–GFP, associated with cis elements, P ars– O ars and P cad– O cad, respectively and dissociated from those upon recognition of As(III) or Pb/Cd. Cell lysates containing ArsR–GFP were pre-incubated with As(III) standard solutions for 15 min and loaded into P ars– O ars-immobilized microplate wells. Cell lysates containing CadC–GFP were pre-incubated with Pb or Cd solutions and loaded into P cad– O cad-immobilized wells. The cell lysates were incubated for 15 min and removed from the wells. Fluorescence intensity in the wells dose-dependently decreased in response to As(III) up to 200 μg/l or Pb/Cd up to 100 μg/l. Detection limits were 10 μg/l for As(III) 10 μg/l for Cd, and 20 μg/l for Pb with a microplate fluororeader, whereas 5.0 μg/l for As(III), 1.0 μg/l for Cd, and 10 μg/l for Pb with a handheld fluorometer. This method was available to detect Pb/Cd or As(III) in water containing soil extracts. This is the first demonstration of a simple and rapid fluorometry to detect analytes based on in vitro interaction between a cis element and a trans factor.