The Effect of Adrenaline and Trypan Blue Used During Cataract Surgery on Anatomical and Functional Outcomes in Pseudoexfoliation Syndrome Patients.

PURPOSE: To evaluate the effect of the intracameral adrenaline and trypan blue used during cataract surgery on corneal endothelial parameters in pseudoexfoliation syndrome (PEX) patients. METHODS: The patients were divided into four groups according to intraoperative use of agents during cataract surgery: intracameral adrenaline (1/10,000, 0.1 ml) (group 1), trypan blue (0.6 mg/ml, 0.1 ml) (group 2), combination of adrenaline and trypan blue (group 3) and none (group 4). RESULTS: Preoperative ECD, CV, HEX and CCT parameters were similar between the groups. A mean loss of 12.7% in ECD was observed at the postoperative third months compared to the preoperative. In group 3, ECD was found to be lower in the postoperative third months compared to the preoperative (p = 0.014). In the other groups, no statistically significant difference was found in preoperative and postoperative comparisons. CONCLUSION: In conclusion, the utilization of intracameral adrenaline or trypan blue did not cause a significant difference in corneal endothelium in PEX patients. However, their combined use may have a negative effect on endothelial cell density. In a cataract surgery performed in the presence of PEX, the increased likelihood of endothelial damage should be taken into consideration, and appropriate precautions should be taken preoperatively and intraoperatively.

In Vivo Tracking and 3D Mapping of Cell Death in Regeneration and Cancer Using Trypan Blue.

Tracking cell death in vivo can enable a better understanding of the biological mechanisms underlying tissue homeostasis and disease. Unfortunately, existing cell death labeling methods lack compatibility with in vivo applications or suffer from low sensitivity, poor tissue penetration, and limited temporal resolution. Here, we fluorescently labeled dead cells in vivo with Trypan Blue (TBlue) to detect single scattered dead cells or to generate whole-mount three-dimensional maps of large areas of necrotic tissue during organ regeneration. TBlue effectively marked different types of cell death, including necrosis induced by CCl4 intoxication in the liver, necrosis caused by ischemia-reperfusion in the skin, and apoptosis triggered by BAX overexpression in hepatocytes. Moreover, due to its short circulating lifespan in blood, TBlue labeling allowed in vivo "pulse and chase" tracking of two temporally spaced populations of dying hepatocytes in regenerating mouse livers. Additionally, upon treatment with cisplatin, TBlue labeled dead cancer cells in livers with cholangiocarcinoma and dead thymocytes due to chemotherapy-induced toxicity, showcasing its utility in assessing anticancer therapies in preclinical models. Thus, TBlue is a sensitive and selective cell death marker for in vivo applications, facilitating the understanding of the fundamental role of cell death in normal biological processes and its implications in disease.

From Ficus recemosa Leaf Galls to Therapeutic Silver Nanoparticles: Antibacterial and Anticancer Applications.

The synthesis of silver nanoparticles (AgNPs) using environmentally friendly methods has become increasingly important due to its sustainability and cost-effectiveness. This study investigates the green synthesis of AgNPs using gall extracts from the plant Ficus recemosa, known for its high phytochemical content. The formation of AgNPs was verified through multiple analytical techniques, including UV-Vis spectroscopy, Fourier-transform infrared (FTIR) spectroscopy, transmission electron microscopy (TEM), zeta potential analysis, and dynamic light scattering (DLS). The UV-Vis spectroscopy results displayed a distinct surface plasmon resonance peak indicative of AgNP formation. FTIR analysis revealed specific interactions between silver ions and phytochemicals in the gall extract, while TEM images confirmed the nanoscale morphology and size of the synthesized particles. Zeta potential and DLS analyses provided insights into the stability and size distribution of the AgNPs, demonstrating good colloidal stability. Biological properties of the AgNPs were assessed through various assays. Antimicrobial activity was tested using the disc diffusion method against Escherichia coli and Staphylococcus aureus, showing significant inhibitory effects. The anticancer potential was evaluated using the trypan blue exclusion assay on Dalton's Lymphoma Ascites (DLA) cells, revealing considerable cytotoxicity. Additionally, antimitotic activity was studied in the dividing root cells of Allium cepa, where the AgNPs significantly inhibited cell division. This research highlights the effective use of F. recemosa gall extracts for the green synthesis of AgNPs, presenting an eco-friendly approach to producing nanoparticles with strong antimicrobial, anticancer, and antimitotic properties. The promising results suggest potential applications of these biogenic AgNPs in medical and agricultural sectors, paving the way for further exploration and utilization.

Combined Brilliant and Trypan Blue As Vital Dyes for Chromovitrectomy.

Brilliant blue 0.05% and trypan blue 0.1% were mixed in a proportion of 1:1 in a 1-mL syringe. This combination produced a waterfall effect with the fast sinking of the dye to the posterior pole and little diffusion through the vitreous cavity. Therefore, it can effectively stain the internal limiting membrane and the epiretinal membrane with a good contrast during surgeries for a macular hole, myopic foveoschisis, and macular pucker.

Canine corneal endothelial cell analysis using vital dyes and light microscopy.

PURPOSE: To evaluate the use of vital dyes and light microscopy for assessing canine corneal endothelial morphology ex vivo. METHODS: The corneas of 40 canine eyes (n = 20 dogs) enucleated <24 h following euthanasia or death were isolated and flat-mounted on a slide. Corneal endothelium was stained via 0.25% trypan blue followed by 0.5% alizarin red (pH 4.2), photographed, then the following morphological features were calculated using ImageJ: mean cell density (MCD), mean cell area (MCA), polymegathism (coefficient of variation of cell area), and pleomorphism (% hexagonality). RESULTS: Mean ± standard deviation (range) outcomes were: MCD, 2544 ± 541 cells/mm2 (1750-3922 cells/mm2); MCA, 431 ± 97 µm2 (251-626 µm2); polymegathism, 17 ± 2% (14%-22%); pleomorphism, 84 ± 3% (80%-90%). No significant differences (p ≥ .122) were noted for any outcome between male versus female or brachycephalic versus non-brachycephalic dogs. Young dogs (<10 years) had lower MCA (p = .044), lower pleomorphism (p = .003), and higher MCD (p = .035) when compared to older dogs (≥10 years). Age was significantly (p ≤ .049) correlated with MCA (r = 0.467), MCD (r = -0.476), polymegathism (r = 0.444), and pleomorphism (r = 0.609). CONCLUSIONS: The combination of vital dyes and light microscopy allowed for clear visualization and evaluation of the corneal endothelium in canine eyes ex vivo. Our findings can be used in future studies to deepen our understanding of the corneal endothelium in health and disease.

Formulation of silver phosphate/graphene/silica nanocomposite for enhancing the photocatalytic degradation of trypan blue dye in aqueous solution.

Photocatalytic degradation of several harmful organic compounds has been presented as a potential approach to detoxify water in recent decades. Trypan Blue (TB) is an acidic azo dye used to distinguish live cells from dead ones and it's classified as a carcinogenic dye. In this study, silver phosphate (Ag3PO4) nanoparticles and novel Ag3PO4/graphene/SiO2 nanocomposite have been successfully prepared via simple precipitation method. Afterward, their physical properties, chemical composition, and morphology have been characterized using SEM, EDS, TEM, SAED, BET, XRD, FTIR and UV-VIS spectroscopy. The specific surface area of Ag3PO4 and Ag3PO4/G/SiO2 nanocomposite were reported to be 1.53 and 84.97 m2/g, respectively. The band gap energy of Ag3PO4 and Ag3PO4/G/SiO2 nanocomposite was measured to be 2.4 and 2.307 eV, respectively. Photocatalytic degradation of Trypan blue (TB) was studied at different parameters such as pH, catalyst dosage, initial concentration, and contact time. The results showed that, at initial dye concentration of 20 ppm, pH = 2, and using 0.03 g of Ag3PO4/G/SiO2 as a photocatalyst, the degradation percent of TB dye in the aqueous solution was 98.7% within 10 min of light exposure. Several adsorption isotherms such as Langmuir, Freundlich, and Temkin adsorption isotherms have been tested in addition to the photocatalytic degradation kinetics. Both catalysts were found to follow the Langmuir isotherm model and pseudo-second-order kinetic model. Finally, the possible photocatalytic performance mechanism of Ag3PO4/G/SiO2 was proposed.

Evaluation of viability, developmental competence, and apoptosis-related transcripts during in vivo post-ovulatory oocyte aging in zebrafish Danio rerio (Hamilton, 1822).

Introduction: Post-ovulatory aging is a time-dependent deterioration of ovulated oocytes and a major limiting factor reducing the fitness of offspring. This process may lead to the activation of cell death pathways like apoptosis in oocytes. Methodology: We evaluated oocyte membrane integrity, egg developmental competency, and mRNA abundance of apoptosis-related genes by RT-qPCR. Oocytes from zebrafish Danio rerio were retained in vivo at 28.5°C for 24 h post-ovulation (HPO). Viability was assessed using trypan blue (TB) staining. The consequences of in vivo oocyte aging on the developmental competence of progeny were determined by the embryo survival at 24 h post fertilization, hatching, and larval malformation rates. Results: The fertilization, oocyte viability, and hatching rates were 91, 97, and 65% at 0 HPO and dropped to 62, 90, and 22% at 4 HPO, respectively. The fertilizing ability was reduced to 2% at 8 HPO, while 72% of oocytes had still intact plasma membranes. Among the apoptotic genes bcl-2 (b-cell lymphoma 2), bada (bcl2-associated agonist of cell death a), cathepsin D, cathepsin Z, caspase 6a, caspase 7, caspase 8, caspase 9, apaf1, tp53 (tumor protein p53), cdk1 (cyclin-dependent kinase 1) studied, mRNA abundance of anti-apoptotic bcl-2 decreased and pro-apoptotic cathepsin D increased at 24 HPO. Furthermore, tp53 and cdk1 mRNA transcripts decreased at 24 HPO compared to 0 HPO. Discussion: Thus, TB staining did not detect the loss of oocyte competency if caused by aging. TB staining, however, could be used as a simple and rapid method to evaluate the quality of zebrafish oocytes before fertilization. Taken together, our results indicate the activation of cell death pathways in the advanced stages of oocyte aging in zebrafish.

Synergetic impact of MgO on PMMA-ZrO2 hybrid composites: Evaluation of structural, morphological and improved mechanical behavior for dental applications.

This work aims to demonstrate the effect of ZrO2 and MgO inclusion into the Poly(methyl methacrylate) (PMMA). To fabricate novel hybrid composites via heat cure method, various composites (PZM2, PZM4 and PZM6) were synthesized in the system [(95-x) PMMA + 5 ZrO2 + x MgO] (x = 2, 4, and 6) respectively. Density of the prepared composites were determined and varying between 1.035-1.152 g/cm3. X-ray Diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) followed by EDAX and mechanical testing were performed to evaluate the fabricated composite properties. Moreover, to explore the structure of the fabricated composites the 13 C CP-MAS SSNMR and 1 H-13 C Phase-Modulated Lee Goldberg (PMLG) HETCOR Spectrum were recorded which clarify chemical shifting and motional dynamics of the composites. Mechanical tests were performed by UTM and the obtained parameters such as compressive strength, Young's modulus, fracture toughness, brittleness coefficient, flexural strength and flexural modulus are found to be in the range of 91-100 MPa, 0.48-0.51 GPa, 9.122-9.705 MPa.m1/2, 0.66-0.815, 51.03-42.78 MPa and 499-663 MPa respectively. Some more mechanical parameters such as proportional limit, elastic limit, failure strength, modulus of resilience and modulus of toughness were also calculated. Furthermore, tribological properties were also determined and the coefficient of friction (COF) was decreased by 17.4 % and 38 % for composite PZM6 at 20 N and 40 N as compared to the composite PZM2 and the lowest wear volume of 1.55 mm3 was observed for PZM2, whereas the maximum volume loss of 5.64 mm3 is observed for composite PZM6. To check out the biocompatibility, cytotoxicity and genotoxicity of the fabricated composites the Trypan-blue assay was also performed for PZM2 and PZM6 composites. Dissection on the gut of larvae was also performed on the both composites followed by DAPI and DCFH-DA staining. Therefore, these synthesized samples can be used for the fabrication of denture materials.

Comparison of Techniques: Inverted Flap and Conventional Internal Limiting Membrane Removal in Idiopathic Macular Hole Surgery

AIM: To compare functional and anatomical outcomes between the inverted flap technique and conventional removal of the internal limiting membrane (ILM) in the surgical management of idiopathic macular hole (IMH). MATERIAL AND METHODS: We retrospectively evaluated the anatomical and functional results in 67 eyes of 65 patients operated on for IMH. The patients were operated on either using the conventional ILM peeling technique (first group) or with the inverted ILM flap technique (second group). 43 eyes of 41 patients were included in the first group, 24 eyes of 24 patients in the second group. We indicated for surgery only patients with IMH stage 2-4 according to the Gasse classification. Best corrected visual acuity (VA) was always determined before and two months after surgery. Furthermore, a comparison of both techniques was made according to the average letter gain after surgery, and the effect of surgery was evaluated using OCT with regard to whether IMH closure succeeded. For both techniques, 25G PPV with SF6 tamponade was performed. RESULTS: Hole closure took place in 41 eyes with conventional ILM removal. In one eye, the hole did not close even after reoperation with the same technique. Median ETDRS letter gain was 7.0. VA remained the same in 2 eyes (4.7%), worsened in 7 cases (16.2%), and improved in all other cases (79.0%). In 16 eyes (37.2%), VA improved by 2 or more lines of ETDRS charts. Using the inverted flap technique, the hole was closed in all 24 monitored eyes. Median ETDRS letter gain was 9.5. VA remained the same in 2 eyes (8.3%), worsened in 2 cases (8.3%), and improved in all other cases (83.3%). In 12 eyes (50.0%), VA improved by 2 or more lines of ETDRS charts. There were no serious complications intraoperatively or postoperatively. CONCLUSION: Our study demonstrated the safety and efficacy of both methods. Although the results were not statistically significant, the inverted flap technique recorded a greater ETDRS letter gain (9.5 vs. 7.0) and proportion of closed holes (100% vs. 95.3%) compared to the conventional ILM peeling technique in our set of eyes.

Single-Cell MTT: A Simple and Sensitive Assay for Determining the Viability and Metabolic Activity of Polyploid Giant Cancer Cells (PGCCs).

Solid tumors and tumor-derived cell lines commonly contain highly enlarged (giant) cancer cells that enter a state of transient dormancy (active sleep) after they are formed, but retain viability, secrete growth promoting factors, and exhibit the ability to generate rapidly proliferating progeny with stem cell-like properties. Giant cells with a highly enlarged nucleus or multiple nuclei are often called polyploid giant cancer cells (PGCCs). Although PGCCs constitute only a subset of cells within a solid tumor/tumor-derived cell line, their frequency can increase markedly following exposure to ionizing radiation or chemotherapeutic drugs. In this chapter we outline a simple and yet highly sensitive cell-based assay, called single-cell MTT, that we have optimized for determining the viability and metabolic activity of PGCCs before and after exposure to anticancer agents. The assay measures the ability of individual PGCCs to convert the MTT tetrazolium salt to its water insoluble formazan metabolite. In addition to evaluating PGCCs, this assay is also a powerful tool for determining the viability and metabolic activity of cancer cells undergoing premature senescence following treatment with anticancer agents, as well as for distinguishing dead cancer cells and dying cells (e.g., exhibiting features of apoptosis, ferroptosis, etc.) that have the potential to resume proliferation through a process called anastasis.

Trypan Blue and Endoillumination-Assisted Phacoemulsification in a Patient With Advanced Keratoglobus.

Keratoglobus is a rare subset of noninflammatory corneal ectasia, which is a group of disorders characterized by corneal thinning, projection, and scarring. Patients with keratoglobus commonly present with poor vision. A case of advanced keratoglobus was managed by a modified phacoemulsification surgical technique using endoillumination and capsular staining with trypan blue. In this case, we present a 54-year-old man with keratoglobus. In January 2023, a modified phacoemulsification surgical technique using endoillumination was described with a video in a patient with bilateral corneal opacification, neovascularization, significant peripheral thinning, and moderate to severe corneal opacity in which cataract surgery had to be performed alone without considering penetrating keratoplasty. Postoperatively, the patient was doing well with no leaks. We may conclude that this method allows for better visualization during surgery and decreases the risk of intraoperative complications due to poor visualization in patients with severe corneal opacity.

Molecular docking analysis of chlorpyrifos at the human α7-nAChR and its potential relationship with neurocytoxicity in SH-SY5Y cells.

The organophosphate insecticide chlorpyrifos (CPF), an acetylcholinesterase inhibitor, has raised serious concerns about human safety. Apart from inducing synaptic acetylcholine accumulation, CPF could also act at nicotinic acetylcholine receptors, like the α7-isoform (α7-nAChR), which could potentially be harmful to developing brains. Our aims were to use molecular docking to assess the binding interactions between CPF and α7-nAChR through, to test the neurocytotoxic and oxidative effects of very low concentrations of CPF on SH-SY5Y cells, and to hypothesize about the potential mediation of α7-nAChR. Docking analysis showed a significant binding affinity of CPH for the E fragment of the α7-nAChR (ΔGibbs: -5.63 to -6.85 Kcal/mol). According to the MTT- and Trypan Blue-based viability assays, commercial CPF showed concentration- and time-dependent neurotoxic effects at a concentration range (2.5-20 µM), ten-folds lower than those reported to have crucial effects for sheer CPF. A rise of the production of radical oxygen species (ROS) was seen at even lower concentrations (1-2.5 µM) of CPF after 24h. Notably, our docking analysis supports the antagonistic actions of CPF on α7-nAChR that were recently published. In conclusion, while α7-nAChR is responsible for neuronal survival and neurodevelopmental processes, its activity may also mediate the neurotoxicity of CPF.

Effects of different cryopreservation parameters on the differences between trypan blue and fluorescent SYTO 13/GelRed assays.

Post-thaw cell viability assessment is very important in cryopreservation because it is the main assessment method used to optimize cryopreservation protocols for each cell type; hence, having standardized accurate, quick, and reliable assays for post-thaw cell viability measurements is of utmost importance. The trypan blue exclusion assay and nucleic-acid-binding fluorescence-based assays are two different methods for cell viability assessment. Both assays identify cells with damaged membranes by whether they let a compound enter the cell. In this study, these two assays are compared in the context of cryopreservation and the impacts of important cryopreservation parameters on the differences in measurements are investigated. H9c2 myoblasts were cryopreserved with different freezing protocols. Cell membrane integrities were measured immediately after thaw as well as after cryoprotectant removal by a hemocytometer-based trypan blue dye exclusion assay and a dual fluorometric SYTO 13/GelRed assay; and the results were compared. This study quantifies how (i) the absence or presence of different cryoprotectants, (ii) different cell-cryoprotectant incubation conditions, and (iii) the presence or removal of cryoprotectants after thaw affect the differences between these two viability assays.

Identification of Sarcocystis spp. in Slaughtered Sheep from Spain and Evaluation of Bradyzoite Viability after Freezing.

Sarcocystis spp. are complex apicomplexan parasites that cause a substantial economic impact on livestock used for meat production. These parasites are present worldwide. Our study aimed to identify Sarcocystis species affecting sheep meat in southern-central Spain and to evaluate the effectiveness of freezing for parasite inactivation. A total of 210 condemned samples of sheep meat were thoroughly assessed grossly and microscopically; the presence of macro- and microcysts was confirmed. The samples were then frozen at -20 °C for various time intervals (24, 48, 72, 96, 120, and 144 h) and compared with untreated samples. Bradyzoites were isolated through pepsin digestion for subsequent molecular analysis and viability assessment, employing trypan blue and double fluorescence staining techniques. Our measurements confirmed the presence of S. tenella, S. gigantea, and S. medusiformis in Spanish domestic sheep. Freezing for 96 to 144 h resulted in a significant reduction in parasite viability, with a robust correlation observed between the two staining methods. Both stains effectively measured the viability of Sarcocystis, thereby promising future advances in meat safety.

Iron oxide nanoparticles synthesized usingMentha spicataextract and evaluation of its antibacterial, cytotoxicity and antimigratory potential on highly metastatic human breast cells.

Iron oxide nanoparticles (Fe2O3NPs) were synthesized utilizingMentha spicatasourced from Cyprus as a stabilizing agent. The study delved into assessing the antimicrobial, cytotoxic, anti-proliferative, and anti-migratory potential of Fe2O3 NPs through disc diffusion, trypan blue, and 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay, respectively. Characterization of the synthesized Fe2O3 NPs was conducted using Fourier-transform infrared spectroscopy (FTIR), x-ray diffraction (XRD), UV-vis spectroscopy (UV-vis), scanning electron microscopy (SEM), and energy-dispersive x-ray spectroscopy (EDX). The FTIR, XRD, and SEM-EDX spectra confirmed the successful formation of Fe2O3 NPs. The analysis of UV-vis spectra indicates an absorption peak at 302 nm, thereby confirming both the successful synthesis and remarkable stability of the nanoparticles. The nanoparticles exhibited uniform spherical morphology and contained Fe, O, and N, indicating the synthesis of Fe2O3NPs. Additionally, the Fe2O3NPs formed through biosynthesis demonstrated antimicrobial capabilities againstEscherichia coliandBacillus cereus. The significant anti-migratory potential on MDA-MB 231 human breast cancer cells was observed with lower concentrations of the biosynthesized Fe2O3NPs, and higher concentrations revealed cytotoxic effects on the cells with an IC50of 95.7µg/ml. Stable Fe2O3NPs were synthesized usingMentha spicataaqueous extract, and it revealed antimicrobial activity onE. coliandB. cereus, cytotoxic, anti-proliferative and anti-migratory effect on highly metastatic human breast cancer cell lines.

In vitro Safety Assessment of Extracts and Compounds From Plants as Sunscreen Ingredients.

This work investigated the safety of extracts obtained from plants growing in Colombia, which have previously shown UV-filter/antigenotoxic properties. The compounds in plant extracts obtained by the supercritical fluid (CO2) extraction method were identified using gas chromatography coupled to mass spectrometry (GC/MS) analysis. Cytotoxicity measured as cytotoxic concentration 50% (CC50) and genotoxicity of the plant extracts and some compounds were studied in human fibroblasts using the trypan blue exclusion assay and the Comet assay, respectively. The extracts from Pipper eriopodon and Salvia aratocensis species and the compound trans-ß-caryophyllene were clearly cytotoxic to human fibroblasts. Conversely, Achyrocline satureioides, Chromolaena pellia, and Lippia origanoides extracts were relatively less cytotoxic with CC50 values of 173, 184, and 89 µg/mL, respectively. The C. pellia and L. origanoides extracts produced some degree of DNA breaks at cytotoxic concentrations. The cytotoxicity of the studied compounds was as follows, with lower CC50 values representing the most cytotoxic compounds: resveratrol (91 µM) > pinocembrin (144 µM) > quercetin (222 µM) > titanium dioxide (704 µM). Quercetin was unique among the compounds assayed in being genotoxic to human fibroblasts. Our work indicates that phytochemicals can be cytotoxic and genotoxic, demonstrating the need to establish safe concentrations of these extracts for their potential use in cosmetics.

Comparison of methods for determining Batrachochytrium dendrobatidis zoospore viability.

The emerging fungal pathogen Batrachochytrium dendrobatidis (Bd) threatens hundreds of amphibian species globally. During laboratory-based experiments it is often essential to quantify live Bd cells, but a comparison of the effectiveness of methods for counting and assessing the viability of the infectious zoospore life stage has not been done. A direct comparison of staining methods that assess viability will ensure that the most accurate and efficient method is used. Here, we compared the use of 2 relatively cheap common stains, trypan blue and methylene blue, and assessed their accuracy and precision for estimating the viability of Bd zoospores during both manual counting and colorimetric assays. We stained known proportions of killed Bd zoospores (0, 0.25, 0.50, 0.75, and 1.00) with each stain and estimated the proportion of stained (dead) and unstained (viable) cells in each sample using both manual counting and colorimetric assays. Trypan blue was found to be a much more effective stain than methylene blue for both microscopy and colorimetric assays. Additionally, counting zoospores via microscopy was both a more accurate and precise technique. We recommend using manual counts via microscopy using the trypan blue stain for assessing Bd zoospore viability.

Selective Optical Imaging for Detection of Bacterial Biofilms in Tissues.

SIGNIFICANCE: The development of an imaging technique to accurately identify biofilm regions on tissues and in wounds is crucial for the implementation of precise surface-based treatments, leading to better patient outcomes and reduced chances of infection. AIM: The goal of this study was to develop an imaging technique that relies on selective trypan blue (TB) staining of dead cells, necrotic tissues, and bacterial biofilms, to identify biofilm regions on tissues and wounds. APPROACH: The study explored combinations of ambient multi-colored LED lights to obtain maximum differentiation between stained biofilm regions and the underlying chicken tissue or glass substrate during image acquisition. The TB imaging results were then visually and statistically compared to fluorescence images using a shape similarity measure. RESULTS: The comparisons between the proposed TB staining method and the fluorescence standard used to detect biofilms on tissues and glass substrates showed up to 97 percent similarity, suggesting that the TB staining method is a promising technique for identifying biofilm regions. CONCLUSIONS: The TB staining method demonstrates significant potential as an effective imaging technique for the identification of fluorescing and non-fluorescing biofilms on tissues and in wounds. This approach could lead to improved precision in surface-based treatments and better patient outcomes.

The Role of Trypan Blue as a Conventional and Fluorescent Dye for the Diagnosis of Superficial Mycoses by Potassium Hydroxide (KOH) Testing.

Direct microscopic examination of samples using potassium hydroxide (KOH) is a fast, simple, and inexpensive method to confirm clinical suspicion of superficial mycosis. However, the lack of color contrast in this test makes it difficult to separate any fungal structures from artifacts. The sensitivity of the KOH mount technique may be enhanced using both fluorochromes and conventional stains that highlight the fungal structures when observed under fluorescence microscopy and bright-field, respectively. Here we study the possibility of using Trypan Blue (TB), an azo dye which is often used as a live/dead marker, in the diagnosis of superficial mycoses by KOH testing. TB at 0.01% displayed a fluorescent staining pattern similar to that of Calcofluor White (CFW), the conventional cell wall fluorophore. Furthermore, by adjusting the TB concentration to 0.1-0.3%, in addition to maintaining the fluorescent staining pattern, the fungal elements were stained in blue under bright-field microscopy. Thus, we demonstrate for the first time that TB has the unique property as a fungal stain that can be used in both bright-field and fluorescence microscopy for diagnosis of superficial mycoses by direct microscopic examination.

"Dye another day!" - dyes in ophthalmology.

Background: Dyes are substances that are an integral part of ocular procedures and surgeries. In Clinical practice, dyes help in better visualization and aid in diagnoses of ocular surface disorders. In Surgical practice, dyes help in better resolution of the structures that are otherwise naked to the surgeon's eyes. Purpose: To educate ophthalmologists about the importance and uses of dyes. Synopsis: Dyes have become an important part of an ophthalmologists' clinical as well as surgical practice. This video aims at educating the different characteristics, uses, advantages and disadvantages of each dye. Dyes help in identifying the obscure and highlighting the invisible. The indications and contraindications as well as the side effects of each dye are discussed which would help ophthalmologists in the correct usage of these wonder substances. This video will also help the new eye doctors understand and utilize these dyes judiciously which would aid in their learning process and provide better patient care. Highlights: This video highlights the uses, indications, contraindications and side effects of all the dyes used in ophthalmology. Video Link: https://youtu.be/shdV4a6oc20.