Fluorescence Detection Research Articles

Aflatoxin M1 level and risk assessment in milk, yogurt, and cheese in Tabriz, Iran

Aim and BackgroundThe objective of this study was to investigate the presence of aflatoxin M1(AFM1) in milk, yogurt, and cheese samples collected from Tabriz, Iran. Additionally, the study conducted a risk assessment related to the consumption of milk and dairy products within Tabriz city. Study MethodFor this study, 56 samples (raw milk, pasteurized milk, ultra-high temperature milk (UHT), traditional yogurt, pasteurized yogurt, traditional cheese, and pasteurized cheese) were collected randomly in Tabriz from December 2021 to March 2022. The analysis was carried out using liquid chromatography, which was equipped with a fluorescence detector. The estimated dietary intake (EDI) and the hazard index (HI) were calculated for the risk assessment. Results and DiscussionAFM1 was detected in all samples. The highest concentration of AFM1 was observed in traditional cheese (P<0.05). The lowest concentration was observed in UHT milk and there were no significant differences between raw milk and pasteurized yogurt(P<0.05). Moreover, in all samples, the levels of AFM1 were below the maximum limit permitted by Iranian national standards. AFM1 levels in traditional yogurt exceeded the European Union's permissible limit in 25% of the samples. The HI in all samples was less than one for both adult and child consumers, except for milk samples for children, which were more than one and indicated a medium risk. In conclusion, the levels of contamination in milk and dairy products and risk assessment appear not to pose a public health risk to Tabriz consumers.

Modulating the intrinsic fluorescence properties of amoxicillin by sulfur nanodot-based nanoreactor for its direct fluorescence detection

Direct fluorescence detection of amoxicillin (AMX) is highly desired for food and environment safety but remains a formidable challenge because of its low conjugation degree and stabilization effects on excitons. In this work, we proposed a direct fluorometry for AMX through modulating its fluorescence properties by utilizing sulfur nanodots (S-dots) as nanoreactors. S-dots provided both confined carrier and reactants (polyethylene glycol (PEG) and sulfite ions) to modulate the fluorescence properties of AMX, achieving a 277-fold increase on the fluorescence intensities. PEG and SO32- interacted with AMX through both covalent and hydrogen bounds, which stabilized the excited states, decreased the chances of nonradiative path and activated the radiative decay. The ultrasmall sized S-dots also provided confinement carries for promoting the reactivity between AMX and surface function groups. Quantitative detection of AMX in commercial milk samples was realized through direct recording the fluorescence signals, achieving a limit of detection of 159 nM. The reported results provided a new idea to design fluorescence assays for extremely weak emissive molecules, showing great potential in food safety analysis, environmental and healthy monitoring.

Highly effective fluorescence detection of UO22+ ions by an anionic Na/Eu heterometallic metal–organic framework with Lewis basic chelating sites

Uranium is a serious radioactive contaminant that can easily migrate into groundwater and surface water, posing a deadly threat to human health. Despite the development of various materials for detecting uranyl ions in water, rational design and synthesis of new uranium sensors with sensitivity, selectivity, and fast response remain challenging. Herein, we demonstrate a novel strategy for obtaining highly sensitive and selective UO22+ sensors by constructing heterometallic metal–organic framework with both an anionic skeleton and abundant chelating sites. Namely, the first anionic Na/Eu heterometallic metal–organic framework (Na/Eu-MOF) based UO22+ sensor is successfully synthesized by using sodium salt and europium salt as reactants, and abundant ether groups were introduced into its porous structure as chelating Lewis basic sites. Beneficial from the synergistic effect between the anionic skeleton and the chelating sites, Na/Eu-MOF achieved rapid, selective adsorption and fluorescence quenching toward uranyl ion. The detection limits of Na/Eu-MOF in deionized and lake water were determined to be 49.7 nM and 113.0 nM, respectively, which are lower than the maximum concentration of 130.0 nM in drinking water proposed by the United States Environmental Protection Agency, indicating potential environmental applications. The detection capability of Na/Eu-MOF is attributed to the combination system of energy competition and interactions between UO22+ ions and Na/Eu-MOF. This work offers a feasible strategy to construct an anionic heterometallic metal–organic framework with chelating sites for uranyl detection in an aqueous solution.

RGB Trichromatic Whiteness Assessment of Bio Analytical Chromatographic Tool Using Fluorescence for Quantitation of Semaglutide: Application to Pharmaceutical Preparations and Spiked Plasma.

Semaglutide (SEMG) is one of the most widely used and trending medications to treat type II diabetes and obesity.This work aimed to develop a liquid chromatography with spectroflourimetric detection (HPLC-flourimetry) analysis of SEMG in both its tablet dosage form and plasma. The power of fluorescence detection coupled with HPLC proved its capability as a bioanalytical tool to assay SEMG in plasma samples owing to its simplicity and sensitivity which reached below the Cmax of SEMG. Separation was done using a C18 column with mobile phase of acetonitrile and water acidified with orthophosphoric acid (pH 3.5) (1.41 × 10-5M) in isocratic mode in ratio 57:43 and 1mL/min flow rate after extraction using protein precipitation. Detection was carried out at λ excitation of 238nm and λ emission of 416 and 307nm for SEMG and the internal standard, respectively. Evaluation of greenness of the proposed method was done using AGREE (Analytical GREEnness Metric Approach), ComplexGAPI (Complementary Green Analytical Procedure Index) & the new algorithm RGB 12 model (Red-Green-Blue). They showed that these methods can be a greener alternative with acceptable sensitivity for analysis of SEMG. The developed seven min-assay was validated per ICH as well as FDA bio analytical methods' guidelines to prove its applicability for routine sample analysis and future pharmacokinetic studies.

Quantitative High-Throughput and High-Sensitivity Glycoprofile Analysis of Therapeutic Glycoproteins Using HILIC-FLD and ESI-MS.

Protein glycosylation is recognized as a Critical Quality Attribute for the biological and therapeutic activity of many recombinant proteins. Therefore, glycosylation should be monitored rigorously to ensure the desired quality, safety, and potency of monoclonal antibodies and other therapeutic glycoproteins. However, glycans are highly heterogeneous structures in proteins, and this poses a challenge for glycoprofile analysis. Hydrophilic interaction liquid chromatography with fluorescence detection has been recognized as a standard method for separation and quantification of released N-glycans after conventional 2-aminobenzamide labeling, but the sample preparation procedure is time-consuming with low sensitivity and poor reproducibility. Here we present a streamlined 96-well plate format platform for high-throughput and high-sensitivity glycan profiling. By taking advantage of rapid glycoprotein denaturation, enzymatic deglycosylation, highly sensitive fluorescent derivation, and on-matrix glycan cleanup, the reaction time has been shortened significantly to approximately 10min. A detailed workflow including two-dimensional chromatography fractionation, exoglycosidase sequential digestion, mass spectroscopy glycan mass confirmation, and data interpretation is described. Potential pitfalls and precautions to be taken to ensure data accuracy are also discussed in this chapter.

Determination of PAH Contamination in Breast Milk Samples from Hungarian Volunteering Mothers, Using HPLC–FLD

(1) The evidence is mounting that polycyclic aromatic hydrocarbons (PAHs) are a class of hazardous organic compounds with established carcinogenic and toxic properties. Humans may be exposed to PAHs through several different routes, including diet, inhalation, and dermal contact. There is also a possibility that they could transfer into breast milk following maternal exposure, which could potentially endanger breastfeeding infants. (2) The objective of this study was to ascertain the concentration of polycyclic aromatic hydrocarbons (PAHs) in breast milk samples from 50 Hungarian mothers, employing high-performance liquid chromatography with fluorescence detection (HPLC–FLD). An Incremental Life Risk Calculation (IRCL) model estimated the carcinogenic risk to infants. (3) Total PAH concentrations ranged from 0 to 78 ng/mL, with fluorene (5.3 ng/mL), phenanthrene (3.2 ng/mL), and pyrene (2.5 ng/mL) being the most abundant. PAHs were detected in 48 of the 50 samples, with phenanthrene present in 92% of samples. Dibenzo (a,h)anthracene was not detected. (4) According to the model measurements, most of the samples were within acceptable risk levels; however, 2 samples out of 50 posed a higher risk. Statistical analysis of questionnaires completed by the mothers indicated that factors such as diet, residence, and education may influence PAH levels in breast milk.

Determination of 3-hydroxyanthranilic acid in the sweat of healthy older adults

3-Hydroxyanthranillic acid (3HAA) is one of the key metabolites from the tryptophan (TRP) metabolism pathway and is associated with aging, age-associated diseases, and healthy lifespan extension. This study aims to detect 3HAA in the sweat of healthy older adults using simple, high-performance liquid chromatography (HPLC) method. Chromatographic separation using 20 mmol/L sodium acetate, 3 mmol/L zinc acetate, and 7% (v/v) acetonitrile as mobile phase is possible to simultaneous detect 3HAA, KYN, and TRP with UV and fluorescence detection, respectively, under 6 min. This method demonstrated excellent linearity with coefficient of determination (r2) greater than 0.998 for all analytes. The linear range were 0.05–6 µg/mL for TRP, 0.1–6 µg/mL for KYN and 0.2–6 µg/mL for 3HAA. Percentage recoveries from spiked in human sweat ranged from 90 ± 7–101 ± 3 for TRP, 86 ± 1–92 ± 3 for KYN, and 96 ± 1–103 ± 4 for 3HAA. The precision (%RSD) of repeatability and reproducibility is less than 3% and 6%, respectively. This method was used in a cross-sectional study with 81 participants aged 50–79 years, selected randomly from a local primary healthcare hospital’s sampling frame. A detectable amount of 3HAA was observed in all sweat samples, marking the first report of 3HAA presence in human sweat. Additionally, the results revealed that the 3HAA sweat levels increased with age analyzed in three different age groups ranging from 50–59, 60–69, and 70–79. These findings enhance our understanding of sweat profiles and their correlation with aging, potentially further improving early diagnosis, disease monitoring, and development of customized treatment programs for older adults.

Commentary: Use of a Single-Fiber Optical Probe for the Detection of Tumor Fluorescence in High-Grade Glioma.

Commentary: Use of a Single-Fiber Optical Probe for the Detection of Tumor Fluorescence in High-Grade Glioma.

Intrastrand Photo-Crosslinking of 5-Fluoro-2'-O-methyl-4-thiouridine-Modified Oligonucleotides and Its Implication for Fluorescence-Based Detection of DNA Sequences.

DNA photo-crosslinking reactions occur widely in biological systems and are often used as valuable tools in molecular biology. In this article, we demonstrate the application of an oligonucleotide 5-fluoro-2'-O-methyl-4-thiouridine (FSU)-containing probe for the fluorescent detection of specific DNA sequences. The design of the probe was predicated on studies of agents that could adversely affect its efficiency. The most important of these is the intrastrand photo-crosslinking of single-stranded oligodeoxynucleotides bearing FSU. Our research findings indicate that FSU after photoexcitation can react with nonadjacent bases; specifically, it can react with distant thymine and cytosine residues in the chain, forming fluorescent and nonfluorescent intrastrand crosslinks, respectively. In addition, partial photooxidation of the FSU residue to 5-fluorouridine was also observed. The results of the study are significant in terms of the use of FSU-labeled oligonucleotide probes in the fluorescence-based detection of specific DNA sequences because the creation of a fluorescent intrastrand crosslink could produce a false signal. To overcome this problem, replacing thymidine with deoxyuridine in the FSU-labeled oligonucleotide probe is proposed and tested.

Boron-doped carbon dots with time-resolved room temperature phosphorescence for detection of ciprofloxacin hydrochloride and information encryption applications.

Novel boron-doped carbon dots (BCDs) with extended afterglow characteristics were synthesized via a one-step solvothermal method using acrylamide, sulfosalicylic acid, and sodium tetraborate as protective matrices. The presence of boron from sodium tetraborate introduced an empty orbital, allowing it to form a more extended conjugated system with adjacent oxygen atoms, thereby lowering the energy level of the lowest unoccupied molecular orbital in the system. The phosphorescence emission of these BCDs exhibits a red shift over time from 450 to 500 nm. These BCDs have been effectively utilized to produce anti-counterfeit phosphorescent powder. Additionally, the BCDs display optimal fluorescence excitation at 330 nm and optimal emission at 420 nm. They demonstrate a detection limit for ciprofloxacin hydrochloride of 37 nM in the 1-100 µM concentration range and 26 nM in the 100-210 µM range. This fluorescence detection is governed by an inner filter effect. Real sample testing further confirms that these BCDs serve as excellent sensors for ciprofloxacin hydrochloride.

Repurposing expired metformin to fluorescent carbon quantum dots for ratiometric and color tonality visual detection of tetracycline with greenness evaluation

Repurposing expired drugs is highly significant as it prevents the accumulation of pharmaceutical wastes, optimizes resource usage, and reduces environmental harm. Metformin, used for type II diabetes, lowers blood sugar via decreasing liver glucose production and increasing muscle insulin sensitivity. However, the growing number of expired tablets annually necessitates a strategic approach for repurposing or reusing these tablets. Herein, we develop a simple hydrothermal approach for converting expired metformin to valuable fluorescent carbon quantum dots (m-CQDs) with sizes smaller than 10 nm. The m-CQDs displayed a blue emission at 420 nm when excited at 350 nm. Interestingly, when tetracycline (TC) was added, the blue emission diminished, and a greenish-yellow emission emerged at 530 nm. This emission tunning was the resulting of combination of inner filter effect and aggregation induced emission. Leveraging this dynamic emission tunning, we developed both ratiometric-based fluorescence and color tonality-based visual detection methods for detecting TC in pharmaceuticals and wastewater. More importantly, three assessment tools, ComplexMoGAPI, AGREE, and BAGI were used to evaluate the environmental sustainability of the proposed analytical method. The evaluation confirmed the sustainability of the method and the alignment with the greenness guidelines for analytical methods.

Fluorescent Antimicrobial Peptides Based on Nile Red: Effect of Conjugation Site and Chemistry on Wash-Free Staining of Bacteria.

Fluorescent probes for bacterial detection can be obtained by conjugating antimicrobial peptides with fluorescent dyes. However, little is known about the effect of the conjugation site and linker chemistry on staining efficiency. We synthesized three conjugates of the antimicrobial peptide ubiquicidin with the environmentally sensitive fluorophore Nile Red that differed by the attachment site and the chemical composition of the linker. We showed that incorporating fluorophore as a minimalistic non-natural amino acid resulted in a superior probe compared with the typically used bioconjugation approaches. The new peptide-based probe named UNR-1 displayed red fluorescence and enabled robust wash-free staining of Gram-positive and Gram-negative bacteria. The probe exhibited selectivity over mammalian cells and enabled rapid fluorescence detection of bacteria by fluorescence microscopy and flow cytometry in an add-and-read format. Our results may foster the development of next-generation fluorescent AMPs for clinical laboratory diagnostics and medical imaging.

Multimass Analysis of Adeno-Associated Virus Vectors by Orbitrap-Based Charge Detection Mass Spectrometry.

Adeno-associated virus (AAV) vectors have attracted significant attention as the main platform for gene therapy. To ensure the safety and efficacy of AAV vectors when used as gene therapy drugs, it is essential to assess their critical quality attributes (CQAs). These CQAs include the genome packaging status, the size of the genome encapsidated within the AAV capsid, and the stoichiometry of viral proteins (VPs) that constitute the AAV capsids. Analytical methods have been established for evaluating CQAs, such as analytical ultracentrifugation, capillary gel electrophoresis with laser-induced fluorescence detection, and capillary gel electrophoresis using sodium dodecyl sulfate with UV detection. Here, we present a multimass analysis of AAV vectors using orbitrap-based charge detection mass spectrometry (CDMS), a single-ion mass spectrometry. Orbitrap-based CDMS facilitates the quantitative evaluation of the genome packaging status based on the mass distribution of empty and full particles. Additionally, we established a novel method to analyze the encapsidated genome directly without pretreatment, such as protein digestion or heat treatment, and to estimate the stoichiometric variation of VP for the capsid based on the mass distribution constituted by the single peak corresponding to AAV particles. Orbitrap-based CDMS is a distinctive method that allows multiple mass characterizations of AAV vectors with a small sample volume of 20 μL for 1013 cp/mL in a short time (30 min), and it holds the potential to become a new standard method in the assessment of CQAs for AAV vectors.

Enzyme cascade signal amplification platform with immobilized xanthine oxidase on nanozymes exhibiting enhanced POD-like activity for ultrasensitive dual-mode detection of xanthine

Xanthine (XA) metabolic disorders cause many diseases, hence early and precise XA detection is essential. Herein, a dual-mode enzyme cascade nanosensing platform based on Au/FMZIF-rGO/PCN nanomaterial with dual-emissive and high peroxidase-like (POD-like) activity was devised. The rapid electron transfer between Fe, Mn and P elements and the formation of ZIF-rGO/PCN hybrid enhanced POD-like activity. Aggregation-induced emission (AIE) effect improved fluorescence intensity of AuNCs. Furthermore, utilizing the nanozyme to immobilize xanthine oxidase (XOD) also reduced intermediate product diffusion, improving cascade efficiency and platform detection·H2O2 was catalytically decomposed to generate •OH, lowering fluorescence intensity at 627 nm and oxidizing TMB to produce blue oxTMB. So, dual-mode nanosensing platform was constructed with linear ranges for ratio fluorescence and colorimetric detection of XA spanning 0.5–200 μM and 0.5–300 μM, respectively, and corresponding LODs of 0.11 and 0.24 μM. The nanoplatform suggested promising potential for practical clinical detection of XA and other biomarkers.

Self-assembly of S,N-codoped Ce/Cu bimetallic nanoparticles for fluorescence and visual detection of hexavalent chromium.

Ce2(SO4)3 was doped into 4,6-diamino-2-mercaptopyrimidine (DAMP)-encapsulated copper nanoclusters (CuNCs) via a facile, rapid, low-temperature, and green self-assembly synthesis method to obtain fluorescent S,N-codoped Cu/Ce-DAMP nanoparticles (S,N-codoped Cu/CeNPs) for the detection of Cr(VI). The prepared Cu/CeNPs exhibit double emission peaks at 470nm and 610nm. Thefluorescence emission at 610nm is significantly enhanced due to the aggregation-induced emission (AIE) effect, and the quantum yield is as high as 20.19%. The fluorescence emission at 610nm can be selectively quenched by Cr(VI) due to the internal filter effect (IFE) and dynamic quenching, whereas the weak fluorescence at 470nm remains almost stable. On this basis, a fluorescence assay method for Cr(VI) was established, with good linearity in the concentration range 0.5-120µM and a detection limit (LOD) of 134nM. Using a smartphone to take photos of the fluorescence signals changes caused by Cr(VI) rapid visual detectionis achieved with a linear range of 10-130 μM and a LOD of 2.35 μM. The proposed method was successfully applied to the detection of Cr(VI) in actual water samples.

Ultrabright silicon nanoparticles combined with o-phenylenediamine for ratiometric fluorescence and smartphone imaging dual-mode detection of nitrite

Nitrite (NO2−) is widely present in the natural environment and human daily life. Excessive NO2− can cause harm to the environment and human health. Herein, silicon nanoparticles (SiNPs) with a fluorescence quantum yield of up to 70 % were synthesised using a one-pot hydrothermal method and combined with the common and inexpensive o-phenylenediamine (OPD) to achieve the detection of NO2−. Upon the addition of NO2−, the blue fluorescence of the SiNPs was quenched due to static quenching and Förster resonance energy transfer (FRET), while the yellow fluorescence of benzotriazole, the reaction product of OPD and NO2−, was enhanced, resulting in the fluorescence color change from blue to yellow. Based on these phenomena, a ratiometric fluorescence sensor integrated with smartphone imaging technology was developed. This sensor is notable for its portability, cost-effectiveness, and satisfactory detection limits (0.016 μM for ratiometric fluorescence and 1.64 μM for smartphone imaging). Importantly, it demonstrates high reliability and practicability in detecting NO2− in real water and food samples. This broadens the application of SiNPs in the sensing field and introduces new possibilities for NO2− detection in complex sample matrices.

A clenbuterol detection method based on magnetic separation up-conversion fluorescent probe

In this study, a fluorescence detection method combining aptamer-modified up-conversion nanoparticles (UCNPs) and magnetic nanoparticles (MNPs) was developed for detection of Clenbuterol (CLB). The aptamer-modified magnetic NPs captured CLB, which reacted with the aptamer-modified UCNPs and generated a sandwich complex. The aptamer-modified UCNPs acted as a fluorescence source. The MNP-CLB-UCNP complex was retrieved from the solution using an magnetic field, and the fluorescence intensity was detected by fluorescence spectrophotometry with excitation and emission spectra at 980 nm and in the 400–800 nm region, respectively. The results showed that the fluorescence intensity gradually increased with increasing concentrations of CLB with a good specify. The method was highly sensitive for the quantification of CLB, with a limit of detection of 0.304 ng mL−1. The recovery rate of CLB from pork samples ranged from 84 % to 94.87 %. This fluorescence method enables the sensitive, precise, and accurate quantification of CLB residues in pork samples.

CRISPR/Cas13a-mediated visual detection: A rapid and robust method for early detection of Nosema bombycis in silkworms

The sericulture industry faces a significant threat from the Pebrine disease of silkworms, caused by Nosema bombycis. Nonetheless, the current microscopic diagnostic methods can be time-consuming, labor-intensive, and lacking sensitivity and accuracy. Therefore, it is crucial to develop a novel detection approach that is efficient, highly sensitive, and low-cost. In this regard, the CRISPR/Cas system has the potential to be a fast, accurate, and highly specific method of detection. Herein, using a microplate reader, a portable fluorescence detection device, and test strips as signal output tools respectively, we have efficiently developed three rapid and facile visual detection methods for N. bombycis using a CRISPR/Cas13a system with conjugation of Recombinase polymerase amplification (RPA). We evaluated the sensitivity of this combined technology by comparing it with the positive plasmid standard and the genome standard of N. bombycis. Remarkably, the sensitivity of the CRISPR/Cas13a system for N. bombycis positive plasmid standard based on the microplate reader, portable fluorescence detection device, and test strips was 1 copy/μL, 10 copies/μL, and 1 copy/μL, respectively, while for the N. bombycis genome standards, the detection sensitivity was 10 fg/μL, 10 fg/μL, and 1 fg/μL, respectively. In addition, extensive evaluations have demonstrated that the established technology can accurately detect N. bombycis without cross-reactivity with other pathogens, ensuring a specificity rate of 100%. In brief, this study will provide a practical, efficient, and affordable method for early and rapid detection of N. bombycis in various settings.

Fluorescence‐Based Detector Design Principles for Low Vapor Pressure Analytes

AbstractFluorescence‐based sensing is a promising method for detecting trace quantities (vapors) of chemical threats. However, direct detection at standard temperature and pressure of chemicals with low volatilities, such as the salts of illegal drugs, is difficult to achieve. Herein, the development of a testing platform designed to maximize the response from fluorescent material detection of low volatility analytes, using the salts of illicit drugs as exemplars, is described. The challenges encountered in detecting low‐volatility analytes are highlighted, and the hardware solutions employed to overcome them are detailed. The testing platform is composed of a swab heating unit, a sensing chamber, and optical components that enable detection of illicit drugs via a fluorescence quenching mechanism. The swab heating unit facilitates volatilization of the analytes, with the shape of the sensing chamber and its fabrication material optimized to maximize the interaction of the analyte with the sensing element, increasing sensitivity. The detection platform is able to detect trace amounts (down to 30 ng) of (±)‐3,4‐methylenedioxyamphetamine hydrochloride (MDA•HCl), along with other common illicit drug salts such as cocaine hydrochloride (cocaine•HCl), fentanyl•HCl, and methamphetamine•HCl (MA•HCl).

αB-crystallin mini-peptides support corneal healing in vitro and in vivo in rabbit model.

To evaluate if topical use of αB-crystallin mini-peptides supports corneal healing following flap surgery. Cultured corneal cells were treated with fluorescent tagged αB-crystallin mini-peptides to assess its internalization. Cultured corneal cells pre-treated with or without the mini-peptides were exposed to H2O2 and cell viability was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Elongation of neurites of cultured trigeminal neurones was examined following treatment either with αB-crystallin mini-peptides or protein. Cultured trigeminal neurones were pre-treated either with αB-crystallin mini-peptides or crystallin protein and exposed to H2O2 and presence of beading in the dendrites and axons was assessed. Corneal flap surgery was conducted on rabbit cornea and treated topically either with αB-crystallin peptide (0.5 mg/mL thrice daily for 14d) or phosphate-buffered saline (PBS). Corneal healing was evaluated under slit-lamp biomicroscope, mRNA expression of inflammatory cytokines were assessed and the corneas were evaluated by histopathology. Internalization of αB-crystallin mini-peptides was ascertained by the detection of fluorescence within the corneal cells. The MTT assay revealed that treatment with αB-crystallin mini-peptide reduced cell death induced by H2O2 treatment. The mini-peptides did not influence the elongation of trigeminal neurites, but significantly (P<0.05) reduced beading in the neurites. In rabbit eye, the treated corneas showed reduced hyper-reflective zones (P<0.05) and suppression in the expression of inflammatory cytokines. Histopathological examination also revealed reduction of inflammatory response in treated corneas. The αB-crystallin mini-peptides restrict the damage to corneal cells and neurons and aids in corneal healing.