RGB Analysis Research Articles

Analysis of Atmospheric Conditions during the Occurrence of Mesoscale Convective Complex (MCC) in East Kalimantan and Surrounding Area (Case Study: December 26, 2021)

Mesoscale Convective Complex (MCC) was first introduced in 1980 through research on infrared (IR) satellite images by Maddox. The MCC phenomenon can caused rain with a long period of time. This study aims to determine the atmospheric conditions when MCC occurred in East Kalimantan and surroundings area on December 26, 2021. The research data used in this study is Himawari-8 satellite data to display cloud top temperatures and convective cloud distribution using the CCO method and image interpretation using RGB method. Analysis of atmospheric conditions was also carried out on air humidity, vertical velocity, and reflectivity in the atmosphere obtained from weather radar data from the Stasiun Meteorologi Sultan Aji Muhammad Sulaiman - Sepinggan. Based on the results of the analysis, MCC was identified as a mature phase on December 26, 2021 at 12.00 UTC. Airmass RGB analysis shows the presence of clouds which are known as Cb clouds through CCO analysis as well as warm air masses with high humidity in East Kalimantan. Negative value of vertical velocity also indicates an updraft of moist air masses and the maximum reflectivity value of the CMAX product, which is 53 dBz, indicates strong convection activity.

Spectral and RGB analysis of the light climate andits ecological impacts using an all-sky camerasystem in the Arctic.

The ArcLight observatory provides an hourly continuous time series of all-sky images providing light climate data (intensity, spectral composition, and photoperiod) from the Arctic (Svalbard at 79°N). Until recently, no complete annual time series of light climate relevant for biological processes has been provided from the high Arctic because of insufficient sensitivity of commercial light sensors during the Polar Night. The ArcLight set up is unique, as it provides both all-sky images and the corresponding integrated spectral irradiance in the visible part of the solar electromagnetic spectrum (E P A R ). Here we present a further development providing hourly diel-annual dynamics from 2020 of the irradiance partitioned into the red, green, and blue parts of the solar spectrum and illustrate their relation to weather conditions, and sun and moon trajectories. We show that there is variation between the RGB proportions of irradiance throughout the year, with the blue part of the spectrum showing the greatest variation, which is dependent on weather conditions (i.e.,cloud cover). We further provide an example of the biological impact of these spectral variations in the light climate using in vivo Chl a-specific absorption coefficients of diatoms (mean of six low light acclimated northern-Arctic bloom-forming species) to model total algal light absorption (AQ t o t a l ) and the corresponding fraction of quanta used by Photosystem II (AQPSII) (O 2 production) in RGB bands and the potential impacts on the photoreceptor response, suggesting periods where repair and maintenance functions dominate activity in the absence of appreciable levels of red or green light. The method used here can be applied to light climate data and spectral response data worldwide to give localized ecological models of AQ.

Multicolor emission-based nitrogen, sulfur and boron co-doped photoluminescent carbon dots for sequential sensing of Fe3+ and cysteine: RGB color sensor and live cell imaging

Herein, a simple hydrothermal synthesis is used to prepare multiple heteroatom-doped photoluminescent carbon dots (CDs) from thiourea (N and S source) and boric acid (B source) as precursors. The optical and physicochemical properties of the as-synthesized NSB-CDs were studied using UV–Vis, photoluminescence, TEM, FT-IR, XRD, Raman, and XPS analyses. The NSB-CDs exhibited excellent stability, high photostability, pH, and ionic strength tolerance; they retained their excellent stability independent of excitation. The NSB-CDs featured small sizes of approximately 3.2 ± 0.4 nm (range: 2.0–5.0 nm) as evidenced using TEM measurements. The NSB-CDs were used as a photoluminescent sensing platform to detect Fe3+ as well as cysteine (Cys) molecules. The competitive binding of Cys to Fe3+ resulted in NSB-CDs that retained their photoluminescence. For the rapid identification and quantification of Fe3+ and Cys, NSB-CDs were developed as a “switch-on” dual-function sensing platform. The linear detection range of Fe3+ was 0–20 μM (limit of detection [LOD]: 54.4 nM) and that of Cys was 0–50 μM (LOD: 4.9 nM). We also introduced a smartphone RGB analysis method for detecting low-concentration solutions based on digital images. The NSB-CDs showed no toxicity at 100 μg/mL. Photoluminescent probes for multicolor live-cell imaging can be used with NSB-CDs at this concentration, suggesting that NSB-CDs may be promising photoluminescent probes.

Design of CsPbBr3 NCs/MnO2 based sensing system for multi-mode detection of D-penicillamine

Herein, a multi-mode assay platform was constructed for the detection of D-penicillamine based on perovskite nanocrystals (CsPbBr3 NCs) and MnO2 nanosheets. The water-dispersible CsPbBr3 NCs synthesized via aqueous emulsion process has a strong fluorescence emission at 520 nm. The electrostatic interaction between positively charged CsPbBr3 NCs and negatively charged MnO2 nanosheets would lead to the fluorescence quenching of CsPbBr3 NCs on account of the inner filter effect (IFE). The addition of D-penicillamine could convert MnO2 nanosheets to Mn2+ and weaken inner filter effect, resulting in distinct recovery of fluorescence at 520 nm and the decrease of the absorption of CsPbBr3 NCs/MnO2 at 375 nm. Therefore, the fluorescence and colorimetric assay system for D-penicillamine was developed. The fluorescence and colorimetric sensing system could detect D-penicillamine ranging from 0.1 to 40 µM and 0.5 to 40 µM with the limits of detection (LOD) were 0.061 µM and 0.24 µM, respectively. The developed assay system could be applied to analyze D-penicillamine in real samples and acquired satisfactory results. Furthermore, a portable intelligent detection device was developed based on the mobile phone and RGB analysis software, demonstrating the practicability and potential of the sensing system.

Assessing the Surface Properties of Plant Extract-Based Silver Nanoparticle Coatings on 17–4 PH Stainless Steel Foams Using Artificial Intelligence-Supported RGB Analysis: A Comparative Study

Assessing the Surface Properties of Plant Extract-Based Silver Nanoparticle Coatings on 17–4 PH Stainless Steel Foams Using Artificial Intelligence-Supported RGB Analysis: A Comparative Study

Visual Colorimetric Detection of Edible Oil Freshness for Peroxides Based on Nanocellulose.

Traditional methods for evaluating the edibility of lipids involve the use of organic reagents and complex operations, which limit their routine use. In this study, nanocellulose was prepared from bamboo, and a colorimetric reading strategy based on nanocellulose composite hydrogels was explored to monitor the freshness of edible oils. The hydrogels acted as carriers for peroxide dyes that changed color according to the freshness of the oil, and color information was digitized using UV-vis and RGB analysis. The sensitivity and accuracy of the hydrogel were verified using H2O2, which showed a linear relationship between absorbance and H2O2 content in the range of 0-0.5 and 0.5-11 mmol/kg with R2 of 0.9769 and 0.9899, respectively, while the chromatic parameter showed an exponential relationship with R2 of 0.9626. Surprisingly, the freshness of all seven edible oil samples was correctly identified by the hydrogel, with linear correlation coefficients greater than 0.95 in the UV-vis method and exponential correlation coefficients greater than 0.92 in the RGB method. Additionally, a peroxide value color card was established, with an accuracy rate of 91.67%. This functional hydrogel is expected to be used as a household-type oil freshness indicator to meet the needs of general consumers.

Colorimetric and fluorescent dual-signals probes for naked-eye detection of hydrogen peroxide and applications in milk samples and in vivo

Excessive residual hydrogen peroxide (H2O2) disinfectant in food is harmful to human health. Therefore, it is necessary to develop efficient detection methods for H2O2 detection. In this work, we designed and synthesized five D-A molecules 3a-3e by introducing electron-donor substituents (–OCH3 and –CH3) to the electron-acceptor dicyanoisophorone skeleton in order to find out the suitable probes for H2O2 detection. Among them, two promising probes, 3a and 3c, are screened out according to structure-property relationships. Based on the principle of intramolecular charge transfer (ICT), 3a and 3c express colorimetric and fluorescent dual-signals towards H2O2 with low detection limits (0.20 μM and 0.14 μM) and rapid response (within 20 mins). The reaction mechanism between probes and H2O2 is determined by 1H NMR and HRMS. Density functional theory (DFT) calculations are measured to study the regulation mechanism of structure adjustment on probs performance. Furthermore, a smartphone RGB analysis is utilized as a portable platform for the quantitative detection of H2O2 without complicated instruments, indicating a high efficiency and on-site detection method for H2O2. In addition, probes are applied to detect H2O2 in milk samples, HepG-2 cells and zebrafish, suggesting the promising applications in food samples and physiological systems.

Azodye-based colorimetric sensor array for identification of biogenic amines: Food forensics by portable RGB-based signal readout

Biogenic amines (BA’s) are known to be released as a result of decarboxylation of amino acids during the long storage of protein rich food products, and thus subsequently established as spoiled food markers. Consequently, the selective and sensitive detection of these biogenic amines is of utmost importance for food safety and monitoring. In this regard, herein, we demonstrate the azodye-based colorimetric sensor array for a rapid quantitative profiling of these food spoilage markers. The distinct responses towards various biogenic amines were identified and processed through automated multivariate analysis. Qualitative and quantitative determination of target analytes were performed using LDA and PSLR and results obtained showing linear correlation with different concentrations of target analytes with limit of detection values from 1.23 to 6.60 μM. For practical application, we have developed solid state cellulose paper-based sensor array which exhibits colorimetric changes for different biogenic amines; thus, makes it a potential system to conveniently monitor the concentration of analyte under investigation for food safety and monitoring through RGB analysis.

Portable smartphone-assisted RGB-dependent ratiometric sensing platform for the detection of tetrachloro-p-benzoquinone in river samples

Tetrachloro-p-benzoquinone (TC-PBQ) can be harmful to the environment as a pesticide residue, but its analytical detection has been largely neglected. In this work, a smartphone-assisted RGB-dependent TC-PBQ ratiometric sensing platform for rapid recognition is introduced. Here, the red-emitting mercaptopropionic acid-capped CdTe quantum dots (MPA-capped CdTe) as the reaction unit can be quenched by TC-PBQ, while the blue-emitting carbon dots (CDs) as the reference signal remain unchanged. The limit of detection (LOD) of the sensing platform can reach 59 nM in aqueous solution. By constructing an analysis model with the system RBG value, the smartphone-assisted sensor-coated fluorescent test paper can complete the qualitative and quantitative visual detection of TC-PBQ (LOD is 93 nM). The applicability of smartphone-dependent RGB analysis significantly simplifies and speeds up the operation process, which provides a new analytical method for real-time analysis of TC-PBQ in real samples.

HgAu alloy@4H-Chromene predicated colorimetric lateral flow device: On-site rapid detection of hyper and hypocholesterolaemia

An unbalanced diet and lifestyle nowadays are very concerning as it lacks desired nutrition’s which in the long term ultimately led to high cholesterol and other health problems. The present work reports the development of a HgAu alloy stabilized 4H-chromene organic nanoparticles (G4) following a simple condensation technique and characterized via HRTEM, SEM, AFM, DLS, EDAX, and STEM. Furthermore, G4 exhibits increased stability in comparison to bare Hg-Au alloy and ultrahigh selectivity and sensitivity towards cholesterol by changing its color from yellow to pink and blue on report of high and low levels of cholesterol respectively. The color change was quantified using RGB analysis and compared with the UV-Visible absorption data. Further for the rapid on-site detection of cholesterol, pristine paper based lateral flow device has been developed and tested for the human serum, where it gives significantly good results with a recovery percentage of 101.7% with 0.011 nM limit of detection (LOD), linearity of 99.3%, and reproducibility of ± 4% (relative standard deviation). To the best of our knowledge, the present work represents the first report on: (a) preparation of HgAu alloy@4H-chromene, and (b) detection of hypocholesteraemia.

Teaching Microextraction Techniques During COVID-19 Pandemic through Remote Lab Strategy

Since the appearance of the COVID-19 pandemic and its subsequent stage, the changes produced in the chemistry teaching process have allowed educators to use a wide range of strategies, methodologies, and tools to promote student learning. To carry out laboratory practices, some strategies reported include the use of simulators and educational software, as well as the development of remote lab activities. In recent years, the teaching of microextraction techniques and the greening of the sample treatment process represent an important topic in instrumental analytical chemistry courses, incorporating green chemistry topics in the curricula. In this context, a hands-on activity through a kit of materials, equipment, and reagents delivered to students for teaching microextraction techniques during the pandemic’s learning conditions is presented. The activity was designed to extract a food coloring dye applying four microextraction techniques, including solid-phase extraction (SPE), solid-phase microextraction (SPME), in-tube solid-phase microextraction (IT-SPME), and micro polymeric magnetized bar adsorptive extraction (μ-PMBAE). Finally, for quantitative analysis, a colorimetry and absorbance-based experiment through image acquisitions and RGB analysis was used. The experiments were applied as extracurricular activities, and students’ feedback was fruitful due to the enthusiasm that it produced in them to carry out laboratory tasks after so much time of confinement. Among the successful points of the activity, it is possible to mention the ease of carrying out the experiments from home and the application of green sample treatment techniques that minimize the use of materials and reagents.

Portable dual-mode biosensor based on smartphone and glucometer for on-site sensitive detection of Listeria monocytogenes

Contamination of Listeria monocytogenes (L. monocytogenes) in the environment and food can pose a serious threat to human health, and there is an urgent need to establish sensitive on-situ detection methods to mitigate its hazards. In this study, we have developed a field assay that combines magnetic separation technology with antibody-labeled ZIF-8 encapsulating glucose oxidase (GOD@ZIF-8@Ab) to capture and specifically identify L. monocytogenes while GOD catalyzes glucose catabolism to produce signal changes in glucometers. On the other side, horseradish peroxidase (HRP) and 3,3′,5,5′-tetramethylbenzidine (TMB) were added to recombined with the H2O2 generated by the catalyst to form a colorimetric reaction system that changes from colorless to blue. The smartphone software was used for RGB analysis to complete the on-site colorimetric detection of L. monocytogenes. This dual-mode biosensor showed good detection performance for the on-site application of L. monocytogenes in lake water and juice samples, both with a limit of detection up to 101 CFU/mL and a good linear range of 101–106 CFU/mL. Therefore, this dual-mode on-site detection biosensor has a promising application for the early screening of L. monocytogenes in environmental and food samples.

A Qualitative Assessment of River Plumes Coupling SWAT Model Simulations and a Beach Optical Monitoring System

The study of plumes occurring at the mouth of small rivers of temporal flow is a challenging task due to the lack of sedimentological and flow data of appropriate spatiotemporal scales. The present contribution examined the case of a typical un-gauged intermittent Mediterranean stream located in Northern Crete (Xiropotamos river). The SWAT (soil and water assessment tool) model was used to simulate and reproduce the hydrological behavior of the adjacent intermittent (Giofyros) river discharging at the same beach, the basin of which has the same geomorphological and hydrological characteristics. The output of the calibrated SWAT model was used to simulate daily flow data for the year 2014. The results were then considered together with the results of the RGB analysis of optical datasets of high spatio-temporal resolution for the same period, derived from a beach optical monitoring system (BOMS). The RGB analysis of the optical (TIMEX) imagery was shown to be a useful technique to identify and classify coastal plumes by using the spatio-temporal variability of pixel properties. The technique was also shown to be useful for the (qualitative) validation of the SWAT output and could be further improved by the collection of ‘ground truth’ data.

A digital microfluidic platform coupled with colorimetric loop-mediated isothermal amplification for on-site visual diagnosis of multiple diseases.

Traditional diagnosis of infectious diseases relies on advanced analyzers in central hospitals, which is not sufficient for rapid control of epidemics, especially in resource-limited areas, raising the importance of the development of diagnostic systems for point-of-care testing (POCT). Here, we developed a simple and cost-effective digital microfluidic (DMF) platform in combination with colorimetric loop-mediated isothermal amplification (LAMP) for simple on-site diagnosis of diseases by the naked eye. The DMF chip contained four parallel units for the simultaneous detection of multiple genes and samples at a time. After amplification, the results were visualized by endpoint detection with concentrated dry neutral red on the chip. The whole process could be completed within 45 min and the on-chip LAMP reaction was shortened to 20 min. The analytical performance of this platform was evaluated by the detection of Enterocytozoon hepatopenaei, infectious hypodermal and hematopoietic necrosis virus, and white spot syndrome virus genes of shrimp. The DMF-LAMP assay showed a detection limit of 101 copies per μl for each target, exhibiting comparable sensitivity to the conventional LAMP assay but was more efficient. The sensitivity was also competitive with that of microfluidic-based LAMP assays using other POCT devices such as centrifugal discs regarding the detection of the same targets. Moreover, the proposed device possessed a simple chip structure and high flexibility to integrate multiplex analysis, which was beneficial for further widespread use in POCT. The practicability of the DMF-LAMP assay was verified by the testing of field shrimp. The result of the DMF-LAMP assay indicated a good agreement with the qPCR method, with Cohen's kappa values ranging from 0.91 to 1.00 depending on different targets. For the first time, an image processing method was established based on RGB analysis under different lighting conditions, and a universally adaptable positive threshold was determined regardless of lighting conditions. Paired with a smartphone, the objective analytical method was facile to implement in the field. Moreover, the DMF-LAMP system is easy to extend for a broad range of bioassays, with the advantages of low-cost, rapid detection, ease of use, good sensitivity, and easy readout.

A novel metal-organic framework of Co-hemin for portable and visual colorimetric detection of 2,4-dichlorophenoxyacetic acid.

On-site quantitative analysis of 2,4-dichlorophenoxyacetic acid (2,4-D) is of significant importance for addressing increasing concerns about public health and environmental quality. Here, a novel metal-organic framework (MOF) of Co-hemin is synthesized and first used for on-site colorimetric monitoring of 2,4-D. 2,4-D as an inhibitor of alkaline phosphatase could specifically suppress the production of ascorbic acid, which restrained in situ etching of Co-hemin and further triggered the colorimetric response. In the colorimetric assay, Co-hemin displayed good oxidase-like activity without addition of H2O2, which could avoid the shortcomings of H2O2 such as toxicity and instability. The Co-hemin biosensor exhibited a relatively low detection limit of 33 ng mL-1 for 2,4-D by the UV method. Moreover, a smartphone based RGB analysis system for the sensitive detection of 2,4-D was developed, and exhibited a good linear relationship between the RGB model parameter and the concentration of 2,4-D. The operability and accuracy of the Co-hemin biosensor were confirmed by the quantitative determination of 2,4-D in real samples, such as serum and tap water. Also, the Co-hemin based colorimetric biosensor showed good selectivity and specificity. Moreover, the developed assays displayed good application in constructing complex logic gates. This work not only provided a portable and visual platform for on-site monitoring of 2,4-D, but also expanded application prospects in the field of complex biological analysis.

An easy-to-use phosphate triggered zinc-azophenine complex assisted metal extrusion assay: A diagnostic approach for chronic kidney disease and in silico docking studies

Regular monitoring of H2PO4- is inevitable owing to its physiological, chemical as well as environmental significance. Herein, a new and promising azophenine-functionalized Zn2+ complex, CN-1·Zn2+ is rationally designed for the selective and sensitive colorimetric detection (LOD = 67.9 ppb) of monophosphorylated species, H2PO4- based on H2PO4--specific metal-extrusion assay through hypsochromic shift in the UV–vis spectra. The sensing phenomenon has been authenticated via UV–vis, ESI-MS, CV, LSV, 1H NMR studies with theoretical (DFT) substantiation. CN-1·Zn2+ has been exploited towards two-input-two-output logic circuitry fabrication, advantageous for molecular-level information storage. Moreover, TLC strip-based contact mode analysis is beneficial for its on-spot applicability. Smartphone-assisted chromogenic recognition efficiency of CN-1·Zn2+via RGB analysis has been well-correlated with spectrophotometric outcomes, demonstrating reliability of the present method. Phosphate detection efficacy of CN-1·Zn2+ in serum specimens with qualified recovery (90.9–98.9 %) makes it a potential diagnostic tool for monitoring chronic kidney disease (CKD). Additionally, CN-1·Zn2+ exhibited DNA binding activity through non-intercalation mode, examined via UV–vis, fluorescence spectroscopic studies with in silico docking. This is the first Zn2+-complex in azophenine family, effectively employed for smartphone-assisted phosphate detection having promising DNA-binding activity, thereby leaving a sustainable footprint in the domain of supramolecular and bio-medical functional world of applications.

Sensitive acid phosphatase assay based on light-activated specific oxidase mimic activity

Acid phosphatase (ACP) is a key marker in the diagnosis of many diseases. However, exploiting a simple and sensitive sensor for the real-time quantitative analysis of ACP is still challenging. Herein, we attempted to develop a sensitive colorimetric sensing strategy for the detection of ACP based on light-activated oxidase mimic property of carbon dots (CDs). The synthesized CDs were proved to be capable of intrinsic light-activated oxidase mimic activity, which could generate reactive oxygen species to oxidize chromogenic substrate under ultraviolet light stimulation. Interestingly, this light-activated oxidase mimic behavior would be effectively suppressed by the antioxidant ascorbic acid (AA), a product from the hydrolysis of 2-phospho-L-ascorbic acid trisodium (AAP) mediated by ACP. Based on the above property, a facile and sensitive colorimetric sensing method for ACP was developed. Under the optimal conditions, the linear range for ACP 0.1–5.5 U/L, and the detection limit was 0.056 U/L. Compared with conventional nanozyme based ACP assay systems, the catalytic activity of light-activated nanozyme could be conveniently regulated by switching the light on and off, which made it easier to precisely control the extent of the reaction and ensured the accuracy of the assay. In addition, the proposed sensing system would be readout directly by the naked eye or smartphone-based RGB analysis system, and have been successfully applied to analyze diluted in diluted fetal bovine serum and urine samples spiked with ACP. All these results indicated that this approach holds good promise for future applications in clinical analysis and point-of-care (POC) biosensor platforms.

Construction of a ratiometric fluorescent probe for visual detection of urea in human urine based on carbon dots prepared from Toona sinensis leaves and 5-carboxyfluorescein

In this work, pH-sensitive blue fluorescent carbon dots (CDs) with high fluorescence quantum yield (17.24%) were synthesized by hydrothermal method using Toona sinensis leaves and ethylenediamine (EDA) as raw materials. The CDs can detect urea with a limit of detection (LOD) of 6.700 mmol L−1. For more sensitive detection of urea, we constructed a ratiometric fluorescent probe (CDs@5-FAM) using CDs and 5-carboxyfluorescein (5-FAM). The CDs@5-FAM probe can rapidly and sensitively detect urea according to the changes of I514/I405, with LOD as low as 0.014 mmol L−1. Furthermore, with the help of a smartphone and RGB analysis software, urea's visual intelligent detection was realized using a CDs@5-FAM probe. The method proposed in this paper is consistent with the standard method, which indicates that the pH-sensitive ratiometric fluorescent probe CDs@5-FAM is accurate and reliable for practical application. It provides a new way for rapid and visual detection of urea.

Dualistic Fluorescence as Well as Portable Smartphone-Assisted RGB-Relied Sensing Assay for the Ultra-Sensitive Determination of Pendimethalin in Food and Water Samples by AIEE Active Organic Probes.

Herein, a peculiar fluorometric as well as smartphone-assisted RGB-relied sensing assay is introduced for determining pendimethalin (PDM) herbicide contents (in parts per trillion level) based on the anthracene-incorporated pyrimidinone/thione probes (S1 to S4). These compounds offered a unique and impressive aggregation-induced emission enhancement (AIEE) behavior by aggregation in H2O-dimethylformamide medium. Furthermore, these AIEE active compounds were found to display superior selectivity and extraordinary sensitivity for PDM detection via fluorescence quenching response. The extent of quenching degree was found to be linearly varied with the PDM concentration ranging from 0 to 20 nM, with a lower limit of detection of 367.8 pM (103.4 ppt) by S3 nanoaggregates. The detailed investigation revealed that such a high sensitivity of the designed sensor toward PDM is attributable to the existence of dual "photoinduced charge transfer and Förster resonance energy transfer process mechanisms". The Stern-Volmer plots, Job's plot, Benesi-Hildebrand plot, and 1H NMR titrations as well indicated the existence of substantial interactions between the sensor and PDM. The conducted selectivity tests provided distinguishable selectivity for PDM detection over various other insecticides/pesticides as well as other structural nitro analogues. Additionally, the presented sensing assay was also applied to quantify the PDM residues in spiked food (vegetables, fruits, and grains) and water samples. In addition, the sensor-coated fluorescent paper test strips were also fabricated for on-site detection of PDM. The applicability of smartphone-relied RGB analysis significantly streamlined the operation process, speeds up the detection procedure, and also offered a novel methodology for real-time analysis of PDM in real samples.

Fabrication of microtiter plate on paper using 96-well plates for wax stamping.

The online version contains supplementary material available at 10.1007/s10404-022-02606-3.