Biomolecules
·
2021
·Vol. 11
·期 8
·1133
·
IF 4.879
Kevin Adi Kurnia,Fiorency Santoso,Bonifasius Putera Sampurna,Gilbert Audira,Jong-Chin Huang,Kelvin H.-C. Chen,Chung-Der Hsiao
Tail coiling is a reflection response in fish embryos that can be used as a model for neurotoxic analysis. The previous method to analyze fish tail coiling is largely based on third-party software. In this study, we aim to develop a simple and cost-effective method called TCMacro by using ImageJ macro to reduce the operational complexity. The basic principle of the current method is based on the dynamic change of pixel intensity in the region of interest (ROI). When the fish tail is moving, the average intensity is increasing. In time when the fish freeze, the peak of mean intensity is maintaining at a relatively low level. By using the optimized macro settings and excel VBA scripts, all the tail coiling measurement processes can be archived with few operation steps with high precision. Three major endpoints of tail coiling counts, tail coiling duration and tail coiling intervals can be obtained in batch. To validate this established method, we tested the potential neurotoxic activity of Tricaine (methanesulfonate, MS-222) and psychoactive compound of caffeine. Zebrafish embryos after Tricaine exposure displayed significantly less tail coiling activity in a dose-dependent manner, and were comparable to manual counting through the Wilcoxon test and Pearson correlation double validation. Zebrafish embryos after caffeine exposure displayed significantly high tail coiling activity. In conclusion, the TCMacro method presented in this study provides a simple and robust method that is able to measure the relative tail coiling activities in zebrafish embryos in a high-throughput manner.Keywords:ImageJ;chorion;tail coiling;ROI;zebrafish;embryo
研究领域:
发育生物学
·
毒理学
✓ 已验证
DOI
PubMed
高影响因子
Inventions
·
2019
·Vol. 4
·期 4
·65
·
IF 0
Fiorency Santoso,Bonifasius Putera Sampurna,Yu-Heng Lai,Sung-Tzu Liang,Erwei Hao,Jung-Ren Chen,Chung-Der Hsiao
This study aimed to develop a simple and cost-effective method to measure blood flow in zebrafish by using an image-based approach. Three days post fertilization (dpf) zebrafish embryos were mounted with methylcellulose and subjected to video recording for tracking blood flow under an inverted microscope equipped with a high-speed CCD camera. In addition, Hoffman lens was used to enhance the blood cell contrast. The red blood cell movement was tracked by using the TrackMate plug-in in the ImageJ image processing program. Moreover, Stack Difference and Time Series Analyzer plug-in were used to detect dynamic pixel changes over time to calculate the blood flow rate. In addition to blood flow velocity and heart rate, the effect of drug treatments on other cardiovascular function parameters, such as stroke volume and cardiac output remains to be explored. Therefore, by using this method, the potential side effects on the cardiovascular performance of ethyl 3-aminobenzoate methanesulfonate (MS222) and 3-isobutyl-1-methylxanthine (IBMX) were evaluated. MS222 is a common anesthetic, while IBMX is a naturally occurring methylxanthine. Compared to normal embryos, MS222- and IBMX-treated embryos had a reduced blood flow velocity by approximately 72% and 58%, respectively. This study showed that MS222 significantly decreased the heart rate, whereas IBMX increased the heart rate. Moreover, it also demonstrated that MS222 treatment reduced 50% of the stroke volume and cardiac output. While IBMX decreased the stroke volume only. The results are in line with previous studies that used expensive instruments and complicated software analysis to assess cardiovascular function. In conclusion, a simple and low-cost method can be used to study blood flow in zebrafish embryos for compound screening. Furthermore, it could provide a precise measurement of clinically relevant cardiac functions, specifically heart rate, stroke volume, and cardiac output.Keywords:blood flow tracking;ImageJ;zebrafish;IBMX;MS222;cardiovascular function assessment
研究领域:
发育生物学
·
毒理学
✓ 已验证
产品关联
DOI
Journal of Future Foods
·
2025
·Vol. 6
·期 1
·48-54
·
IF 7.2
Xudong Shi,Xingjun Xi,Xiaoqian Tang,Yisheng Chen
Aflatoxin B 1 (AFB 1 ) and deoxynivalenol (DON) are among the most frequently detected mycotoxin in cereals, posing great challenge to human health. This work proposed a cost-efficient and robust planar chromatography method for the rapid screening AFB 1 in millet and buckwheat, as well as DON in wheat, corn, barley, and oats, achieving detection of the two common fungal toxins in cereals. First, the sample preparation was realized by a simple extraction with 10 mL of acetonitrile, both containing 4 g of anhydrous MgSO 4 , 1 g of NaCl, and 25 mg of primary secondary amine adsorbent. After high throughput chromatography, which resulted in the separation of the targeted compound from background noise and derivatization with 10% ethanol solution of AlCl 3 (only for the visualization of DON). The separation results were recorded using a smartphone in a dark chamber equipped with a 366 nm ultraviolet source. To simplify the analysis, the quantification of the graphical outcome was carried out using the open-source software ImageJ, based on quantifying digitalized pixel values. It was demonstrated that the detection sensitivity (AFB 1 : limit of detection (LOD) = 14.2 μg/kg, DON: LOD = 64.3 μg/kg) was adequate to meet the strict maximum residue limit stipulated for both targeted compounds, while calibration curves gave good linearity (AFB 1 : correlation coefficient ( R 2 ) = 0.967 within 1−80 ng/band, DON: R 2 = 0.998 within 50−800 ng/band). Then the method accuracy and precision were evaluated by spiking-recovery experiments, showing that the recovery rates of AFB 1 was within 76.9%−113.6% and those DON within 91.0%−116.5%, with relative standard deviations of less than 4.9% and 5.8% for AFB 1 and DON, respectively. The conceptual validation of this study demonstrates that the high performance thin layer chromatography (HPTLC)-image quantification method has effectively addressed the limitation of HPTLC imaging results being confined to preliminary screening without the capability of precise quantification, thereby bridging the gap between imaging and quantification. This method exhibits promising analytical tool characteristics and is suitable for screening mycotoxins in cereal samples.
✓ 已验证
产品关联
DOI
高影响因子
Biology-Basel
·
2025
·Vol. 14
·期 1
·51
·
IF 3.5
Ferry Saputra,Tzu-Ming Tseng,Franelyne P. Casuga,Yu-Heng Lai,Chih-Hsin Hung,Chung-Der Hsiao
Simple SummaryMany studies have been developed to calculate the blood flow velocity in adult zebrafish. However, the current method requires high-cost equipment a lot of training before it can be replicated. In this study, we provide a simple, cheap, and non-invasive method to enable the measurement of zebrafish blood flow velocity. By applying the same method as the one previously developed for larvae zebrafish, blood flow velocity can be measured and its sensitivity validated by performing an experiment under several ambient temperature conditions, as well as the presence of a toxicant. The applicability of this method was also checked by applying the same method to other fish. Overall, this method provides an easy, cheap, and robust method that can be used by low- to medium-scale laboratories to contribute to knowledge in the relevant fields.Blood flow is an important physiological endpoint to measure cardiovascular performance in animals. Because of their innate transparent bodies, zebrafish is an excellent animal model for assessing in vivo cardiovascular performance. Previously, various helpful methods for measuring blood flow in zebrafish larvae were discovered and developed. However, an optimized method to measure blood flow in adult zebrafish has not been reported. In this paper, the tail fin region was selected as target for blood flow measurements using the Trackmate method, provided by ImageJ platform. Based on power statistic calculations, the aortic vessel at the tail base was selected, and other parameters, such as ambient temperature, were investigated for method standardization, in order to minimize experimental variation. The method was also validated using fenpropathrin and ponatinib, which showed some cardiac alterations in a previous zebrafish study. We also checked the versatility of this method by following the same setup in black tetra and medaka and found that this method performed well. However, our results show that heavy pigmentation, like that found in tiger barb, and overlapping vessels, like those in parrot fish, make it hard for this method to perform well. Overall, an optimized protocol was used for the first time to measure blood flow velocity in adult wild-type zebrafish without the aid of transgenic lines or fluorescent dye.
研究领域:
生物学
·
医学
·
毒理学
·
药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Animals
·
2020
·Vol. 10
·期 9
·1587
·
IF 2.323
Fiorency Santoso,Viacheslav V. Krylov,Agnes L. Castillo,Ferry Saputra,Hong-Ming Chen,Hong-Thih Lai,Chung-Der Hsiao
Simple SummaryWith the advantages of easy culture, body transparency, and high sensitivity to chemical pollution, water fleas have been recognized as a good model for ecotoxicity studies. In this paper, we established ImageJ-based methods to measure cardiovascular performance by evaluating the heart rate and blood flow velocity in three water fleas for the first time. Among the three water fleas,Daphnia magnawas identified as having the most robust heartbeat and blood flow rate and is therefore suitable for ecotoxicity assessment. Many important parameters like heart rate, blood flow rate, stroke volume, ejection fraction, fractional shortening, cardiac output heartbeat regularity can be extracted from videotaping and mathematical calculation. In utilizing those physiological parameters, the potential impacts of ambient water temperature and pesticide pollution on water fleas can be precisely measured.AbstractWater fleas are a good model for ecotoxicity studies, and were proposed for this purpose by the United States Environmental Protection Agency, due to their easy culture, body transparency, and high sensitivity to chemical pollution. Cardiovascular function parameters are usually used as an indicator of toxicity evaluation. However, due to the nature of the heart and blood flow, and the speed of the heartbeat, it is difficult to perform precise heartbeat and blood flow measurements with a low level of bias. In addition, the other cardiovascular parameters, including stroke volume, cardiac output, fractional shortening, and ejection fraction, have seldom been carefully addressed in previous studies. In this paper, high-speed videography and ImageJ-based methods were adopted to analyze cardiovascular function in water fleas. The heartbeat and blood flow for three water flea species,Daphnia magna,Daphnia silimis, andMoinasp., were captured by high-speed videography and analyzed using open-source ImageJ software. We found the heartbeat is species-dependent but not size-dependent in water fleas. Among the three water fleas tested,D. magnawas identified as having the most robust heartbeat and blood flow rate, and is therefore suitable for the ecotoxicity test. Moreover, by calculating the diameter of the heart, we succeeded in measuring other cardiovascular parameters.D. magnawere challenged with temperature changes and a pesticide (imidacloprid) to analyze variations in its cardiovascular function. We found that the heartbeat ofD. magnawas temperature-dependent, since the heartbeat was increasing with temperature. A similar result was shown in the cardiac output parameter. We also observed that the heartbeat, cardiac output, and heartbeat regularity are significantly reduced when exposed to imidacloprid at a low dose of 1 ppb (parts per billion). The blood flow rate, stroke volume, ejection fraction, and fractional shortening, on the contrary, did not display significant changes. In conclusion, in this study, we report a simple, highly accurate, and cost-effective method to perform physiological and toxicological assessments in water fleas.Keywords:water flea;Daphnia;cardiovascular performance;ImageJ
研究领域:
生态毒理学
·
生物学
·
环境科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2024
·
IF 3.8
Shen Liyue,Peng Tao,Dong Jiahui,Liang Zhanwei,Si Jihao,Ye Hua,Xie Jie,Yu Xiaoping,Dai Xinhua
Biomarkers screening is a benefit approach for early diagnosis of major diseases. In this study, magnetic nanoparticles (MNPs) have been utilized as labels to establish a multi-line immunochromatography (MNP-MLIC) for simultaneous detection of carcinoembryonic antigen (CEA), carbohydrate antigen 199 (CA 19–9), and alpha-fetoprotein (AFP) in a single serum sample. Under the optimal parameters, the three biomarkers can be rapidly and simultaneously qualitative screening within 15 min by naked eye. As for quantitative detection, the MNP-MLIC test strips were precisely positioned and captured by a smartphone, and signals on the test and control lines were extracted by ImageJ software. The signal ratio of test and control lines has been calculated and used to plot quantitative standard curves with the logarithmic concentration, of which the correlation coefficients are more than 0.99, and the limit of detection for CEA, CA 19–9, and AFP were 0.60 ng/mL, 1.21 U/mL, and 0.93 ng/mL, respectively. The recoveries of blank serum were 75.0 ~ 112.5% with the relative standard deviation ranging from 2.5 to 15.3%, and the specificity investigation demonstrated that the MNP-MLIC is highly specific to the three biomarkers. In conclusion, the developed MNP-MLIC offers a rapid, simple, accurate, and highly specific method for simultaneously detecting multiple biomarkers in serum samples, which provides an efficient and accurate approach for the early diagnosis of diseases. Graphical Abstract
研究领域:
医学诊断
·
纳米技术
·
生物标志物筛查
·
即时检测
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 7
·1-11
·
IF 6.232
Mao Xiaoxia,Liu Shaowei,Su Benyue,Wang Dejin,Huang Zhan,Li Jian,Zhang Yuanguang
A luminescent metal-organic framework of type Eu(III)-MOF has been fabricated for visual and on-site fluorometric determination of hydrogen peroxide (H 2 O 2 ) via a tablet computer. The maximum excitation and emission peaks of type Eu(III)-MOF were found at λex = 290 nm and λem = 615 nm, respectively. The average length of Eu-MOF is 1.21 ± 0.07 μm. In the presence of the target H 2 O 2 , Fe 2+ is transmitted into Fe 3+ via Fenton reaction, leading to a fluorescence quenching of Eu-MOF. Therefore, visible color change occurred from bright red into colorless. Interestingly, by means of tablet computer’s digital camera and ImageJ software, fluorescent signals were captured and transduced into digital parameters, resulting in a linear relationship between fluorescence intensity and the concentration of H 2 O 2 . As a result, the determination of H 2 O 2 without the aid of complicated instruments is achieved in the range 2.0 μM to 0.2 mM with a detection limit of 1.02 μM. Our approach has been successfully applied to quantify H 2 O 2 in serum, urine, and waste water with good recovery and precision (< 2.5% RSD). Besides, our assay has been exploited for visual detection of H 2 O 2 released from HepG2 cells with the advantages of portability and accuracy. Moreover, the strategy displays acceptable selectivity and stability. Hence, our assay provides an alternative practical method for on-site determination of H 2 O 2 without the need for instruments.
Schematic representation of the synthesis procedure of a luminescent Eu-MOF, which has been successfully applied for on-site detection of H 2 O 2 via Fenton reaction and imaging analysis technique. The method exhibits handheld and accuracy for H 2 O 2 determination, holding the potential for biochemical and clinical applications in remote regions.
研究领域:
分析化学
·
生物化学
·
材料科学
·
临床诊断
✓ 已验证
DOI
PubMed
高影响因子
FOOD CONTROL
·
2022
·
IF 6.652
Gaozhen Li,Fukai Guan,Shancang Zhao,Huihui Xu,Jiashuai Sun,Jingcheng Huang,Qingqing Yang,Yanyin Guo,Xia Sun,Yemin Guo
As an organochlorine pesticide, procymidone has become an increasing serious problem in vegetables in recent years. In order to solve a series of problems such as low sensitivity of the immunological methods, a time-resolved fluorescent immunoassay strip (TRFIAS) was developed with low limit of detection (LOD). Based on the principle of antigen-antibody immune competition, TRFIAS could detect procymidone in vegetables. After the targets being added, the samples were incubated at 37°C for 10 min. TRFIAS was observed under a UV lamp of 365 nm. The results were processed by ImageJ software to complete the qualitative and quantitative analysis of TRFIAS. Under the optimal working conditions, a linear range of TRFIAS was 5–250 ng/mL with a LOD of 0.33 ng/mL and a visual limit of detection (VDL) of 50 ng/mL. And the TRFIAS had good specificity, precision and accuracy. Comparing the results of TRFIAS with those of ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), the two assays were in good agreement. Therefore, TRFIAS has the advantages of high sensitivity, high specificity, simple operation and relatively short assay time. Meanwhile, it shows absolutely great advantages in both quantitative and qualitative analysis.
研究领域:
农业化学
·
分析化学
·
食品科学
✓ 已验证
产品关联
DOI
高影响因子
ACS Applied Materials & Interfaces
·
2019
·
IF 8.456
Rui Jin,Deshuai Kong,Xu Yan,Xu Zhao,Hongxia Li,Fangmeng Liu,Peng Sun,Yuehe Lin,Geyu Lu
Precise on-site profiling of organophosphate pesticides (OPs) is of significant importance for monitoring pollution and estimating poisoning. Herein, we designed a simple and convenient portable kit based on Ag+-responsive hydrogels for accurate detection of OPs. The newly developed hydrogels employed o-phenylenediamine (OPD) and silicon quantum dots (SiQDs) as indicator, which possessed ratiometric response. In this sensor, OPs as inhibitor of acetylcholinesterase prevented the generation of thiocholine, which blocked the formation of metal–polymer with Ag+, further triggered the oxidation of OPD to yield yellow 2,3-diaminophenazine (DAP) with fluorescence emission at 557 nm. The fluorescence intensity of SiQDs (444 nm) was quenched by DAP through inner filter effect (IFE) process, emerging a typical ratiometric response. Interestingly, the ratiometric signal of kit, which was recorded by smartphone’s camera, can be transduced by ImageJ software into the hue parameter that was linearly proportional to the concentration of OPs. The simplicity of portable kit combined with smartphone operation, which possessed high sensitivity (detection limit <10 ng mL–1) and rapid sample-to-answer detection time (45 min) in agricultural sample, indicating that the methodology offered a new sight for portable monitoring of food safety and human health.
研究领域:
分析化学
·
环境科学
·
材料科学
·
食品科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2024
·
IF 8.5
Fan Jiang,Nana Jin,Lei Wang,Siyuan Wang,Yanbin Li,Jianhan Lin
Salmonella screening is essential to avoid food poisoning . A simple, fast and sensitive colorimetric biosensor was elaborately developed for Salmonella detection on a microfluidic chip through limiting air chambers for precise air control, switching rotary valves for accurate fluid selection, a convergence-and-divergence passive micromixer and an extrusion-and-suction active micromixer for efficient fluid mixing, and immune gold@platinum palladium nanocatalysts for effective signal amplification. The mixture of bacteria, immune magnetic nanobeads and nanocatalysts was first rapidly mixed to form nanobead-bacteria-nanocatalyst conjugates and magnetically separated for enrichment. After washing with water, the conjugates were used to catalyze colorless substrate and blue product was finally analyzed using ImageJ for quantifying bacterial concentration. The finger-actuated microfluidic chip enabled designated control of designated fluids in designated places towards designated directions by simple press-release operations on designated air chambers without any external power. Under optimal conditions, this sensor could detect Salmonella at 45 CFU/mL in 25 min.
研究领域:
生物传感器
·
微流控技术
·
纳米技术
·
食品安全
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2020
·
IF 3.594
Yisheng Chen,Caihong Huang,Zhengyu Jin,Xueming Xu,Yina Cai,Yuxiang Bai
Ginkgo biloba was a popular phytomedicine as the essential ingredient of many food supplement. In this work, a bio-inspired HPTLC method was proposed for the visualized screening of nifedipine adulteration in food supplement based on Gingko biloba . To customize the visualization of the analyte by inhibiting bacterium ( Photobacterium phosphoreum ) bioluminescence, the silica gel plate that gives the highest detectability was selected as the stationary phase of separation. Thereafter, the HPTLC procedure (toluene and ethyl acetate 8/2, v/v, as the mobile phase, to a distance of 60 mm) was optimized and standardized as a sample cleanup tool to exclude noises caused by bioactive co-extracts. Then, the separation result fixed on the plate layer was selectively visualized as dark bands against the white background, only after 6 min exposure. To achieve quantitation of the obtained HPTLC images, an enhanced quantitative analysis was optimized via software (ImageJ) facilitated pixel scanning, offered remarkable sensitivity (LOD=24.0 μg/g), linearity ( R 2 =0.9985 within 50–300 ng/band) and precision (RSD<7.2%). Further, the developed method was validated with real samples (recovery rates within 81.0–92.8%). In conjunction with image quantitation, in-situ surface enhance Raman spectroscopy was incorporated into the analysis, using silver nanoparticles as the active substrate and He:Ne 633 nm lamp the incident laser source. The obtained SERS spectra displayed highly characteristic fingerprint patterns, enabling unambiguous confirmation of effect-directed results.
研究领域:
药物分析
·
分析化学
·
食品科学
·
植物化学
✓ 已验证
产品关联
DOI
高影响因子
ANALYST
·
2018
·Vol. 143
·期 12
·2901-2907
·
IF 3.864
Wenyue Xie,Lingli Lei,Meiling Tian,Zeying Zhang,Yingshuai Liu
A high-resolution colorimetric immunoassay platform has been developed based on enzyme-catalyzed multicolor generation and smartphone-assisted signal readout. The multi-color generation is accomplished in this system through the urease-catalyzed urea hydrolysis-induced color change of the pH indicator phenol red, from yellow to orange to red over pH 6.6 to 8.0. The color change is easily tailored by controlling the urease activity via the inhibitor silver ion (Ag+), the amount of which is in turn adjusted by alkaline phosphatase (ALP)-catalyzed ascorbic acid (AA) production. An ALP-linked colorimetric immunoassay is readily realized based on the above urease catalyzed multicolor generation system. Under optimal conditions, a limit of detection (LOD) of 1.73 ng mL−1 and a dynamic range from 0 to 18 ng mL−1 are achieved with rabbit IgG as a model analyte. A colored picture for each test is directly taken using a smartphone and then quantitatively analyzed with the free software ImageJ, eliminating the use of expensive and desktop equipment. The dose-dependent multicolor display is easier to distinguish with the naked eye for qualitative or semiquantitative detection over the traditional one-color system. The developed immunoassay scheme provides a promising platform for on-site testing or applications in resource-poor areas.
研究领域:
分析化学
·
生物传感
·
即时检测
·
移动健康
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）