Optimization of Cultivation Conditions of Native Microalga Scenedesmus quadricauda and Evaluation of Lipids for Enhanced Biodiesel Production.

Biofuels are considered to be among the primary alternatives to the use of fossil fuels. These fuels, made from feedstock or waste raw materials, have the advantage of being renewable and contributing much less to global warming. Microalgae are a promising biodiesel source. Microalgae, unlike traditional crops that are now used to make commercialized biodiesel, may be grown on non-agricultural land and has a greater capacity for growth and yield. Cultivation has been considered as a critical stage in the generation of biofuels. The goal of the present study is to learn that Scenedesmus quadricauda has a potential for biodiesel production in the near future. Optimization studies revealed that BG-11 medium, temperature of 25 °C, pH 7.0, glucose and sucrose (as carbon sources), static condition (for lipid accumulation) & shaking condition (for biomass yield), cultivation days of 18, 21, and 24 day, NaNO3 dosing of 1.0 mM followed by 0.8 mM (on 5th day of cultivation), 3% yeast extract dosing, 3000 lx light intensity, photoperiod cycles of 24L/0D (for biomass yield) and 18L/6D (for lipid production) and 10 mM concentration of NaCl (salinity stress) can be regarded as best suited physio-biochemical parameters for efficient biomass and lipid yield from S. quadricauda. FTIR indicated presence of various stretching of carbohydrates and lipids that again is supporting biodiesel production capability of S. quadricauda. SEM showed that cells of S. quadricauda under stress conditions became fragmented separated from coenobium and were not so compactly arranged. Present optimization studies along with Nile red fluorescence, FTIR and SEM revealed that S. quadricauda could be a suitable candidate to produce good quality biofuel and that also in stress conditions.

A holistic review on red fluorescent graphene quantum dots, its synthesis, unique properties with emphasis on biomedical applications.

Graphene quantum dots (GQDs) are an evolving class of carbon-based nanomaterial, seizing tremendous attention owing to their intense optical property, engineered shapes and structures, and good photostability. Being a zero-dimensional form of carbon structure, GQDs have superior photoluminescent behavior, tunable emission and absorption, excellent biocompatibility, low cytotoxicity, hydrophilic nature, modifying surface states. Their water dispersibility and functionalized surface structure, involving heteroatoms and various functional groups onto the surface of GQDs, make them particularly suitable for biological applications. Based on their absolute luminescence properties, GQDs emit blue, green, yellow, and red light under ultraviolet irradiation. Amongst the three colors, red luminescence can achieve deeper penetration of light into tissues, good cellular distribution, bio-sensing property, cell imaging, drug delivery, and serves as a better candidate for photodynamic therapy. The overall objective of this review is to provide a comprehensive overview of the synthesis methods for red fluorescence graphene quantum dots (RF-GQDs), critical comparative analyses of spectral techniques used for their characterization, the tunable photoluminescence mechanisms underpinning red emission, and the significance of chemically functionalizing GQDs' surface edges in achieving red fluorescence are discussed in depth. This review also discusses the effective biological applications and critical challenges associated with RF-GQDs are examined, providing insights into their future potential in clinical and industrial applications.

Red Fluorescence from Organic Microdots: Leveraging Foldamer-Linked Azobenzene for Enhanced Stability and Intensity in Bioimaging Applications.

Azobenzene, while relevant, has faced constraints in biological system applications due to its suboptimal quantum yield and short-wavelength emission. This study presents a pioneering strategy for fabricating organic microdots by coupling foldamer-linked azobenzene, resulting in robust fluorescence intensity and stability, especially in aggregated states, thereby showing promise for bioimaging applications. Comprehensive experimental and computational examinations elucidate the mechanisms underpinning enhanced photostability and fluorescence efficacy. In vitro and in vivo evaluations disclose that the external layer of cis-azo-foldamer microdots performs a self-sacrificial function during photo-bleaching. Consequently, these red-fluorescent microdots demonstrate extraordinary structural and photochemical stabilities over extended periods. The conjugation of a ß-peptide foldamer to the azobenzene chromophore through a glycine linker instigates a blue-shifted and amplified π*-n transition. Molecular dynamics simulations reveal that the aggregated state of cis-azo-foldamers fortifies the stability of cis isomers, thereby augmenting fluorescence efficiency. This investigation furnishes crucial insights into conceptualizing novel, biologically inspired materials, promising stable and enduring imaging applications, and carries implications for diverse arenas such as medical diagnostics, drug delivery, and sensing technologies.

Fabrication and Efficient Interfacial Assembly of Bright Red-Emitting Carbon Quantum Dots for Security-Warning Textiles.

Carbon quantum dots (CQDs) have attracted more attentions due to their multiple performances. However, the fabrication of long-wavelength emitting CQDs with aliphatic precursors still remains a challenge, mainly because it is difficult to generate large sp2 domains to reduce energy gap, which is not conducive to a redshift of the luminescence peak. Hereon, by regulating the pH of citric acid and thiourea mixture, a N, S co-doped CQD emitting bright red fluorescence at 635 nm is successfully fabricated through the solvothermal reaction under acidic condition, achieving a high quantum yield of 32.66%. Solvatochromic effects of the CQDs are discussed through theoretical equations and models, which confirm that the hydrogen-bonding interaction dominates the fluorescence emission behavior of CQDs in polar solvents. Besides, a feasible strategy is proposed to prepare an anti-counterfeiting textile via the deposition of red-emitting CQDs onto cotton fibers, through rapidly evaporating the preferred organic solvent. As expected, the CQD-decorated textiles exhibit encouraging anti-counterfeiting and security-warning functions, along with underwater and long-distance detectability, washability, and sun resistance. It is worth noting that the present work is innovative in realizing the application of red-light-emitting CQDs in the fields of security-warning textiles.

Nile red staining for rapid screening of plastic-suspect particles in edible seafood tissues.

Concerns regarding microplastic (MP) contamination in aquatic ecosystems and its impact on seafood require a better understanding of human dietary MP exposure including extensive monitoring. While conventional techniques for MP analysis like infrared or Raman microspectroscopy provide detailed particle information, they are limited by low sample throughput, particularly when dealing with high particle numbers in seafood due to matrix-related residues. Consequently, more rapid techniques need to be developed to meet the requirements of large-scale monitoring. This study focused on semi-automated fluorescence imaging analysis after Nile red staining for rapid MP screening in seafood. By implementing RGB-based fluorescence threshold values, the need for high operator expertise to prevent misclassification was addressed. Food-relevant MP was identified with over 95% probability and differentiated from natural polymers with a 1% error rate. Comparison with laser direct infrared imaging (LDIR), a state-of-the-art method for rapid MP analysis, showed similar particle counts, indicating plausible results. However, highly variable recovery rates attributed to inhomogeneous particle spiking experiments highlight the need for future development of certified reference material including sample preparation. The proposed method demonstrated suitability of high throughput analysis for seafood samples, requiring 0.02-0.06 h/cm2 filter surface compared to 4.5-14.7 h/cm with LDIR analysis. Overall, the method holds promise as a screening tool for more accurate yet resource-intensive MP analysis methods such as spectroscopic or thermoanalytical techniques.

Red Emitting Solid-State CDs/PVP with Hydrophobicity for Latent Fingerprint Detection.

Fluorescent carbon dots (CDs) are a new type of photoluminescent nanomaterial. Solid-state CDs usually undergo fluorescence quenching due to direct π-π* interactions and superabundant energy resonance transfer. Therefore, the preparation of solid-state fluorescent CDs is a challenge, especially the preparation of long wavelength solid-state CDs. In this research, long wavelength emission CDs were successfully synthesized by solvothermal methods, and the prepared CDs showed good hydrophobicity. The composite solid-state CDs/PVP (Polyvinyl pyrrolidone) can emit strong red fluorescence, and the quantum yield (QY) of the CDs/PVP powder reaches 18.9%. The prepared CDs/PVP solid-state powder was successfully applied to latent fingerprint detection. The results indicate that the latent fingerprints developed by CDs/PVP powder have a fine definition and high contrast visualization effect, which proves that the prepared CDs/PVP has great application potential in latent fingerprint detection. This study may provide inspiration and ideas for the design of new hydrophobic CDs.

Facile synthesis of Gd/Ru-doped fluorescent carbon dots for fluorescent/MR bimodal imaging and tumor therapy.

Functional metal doping endows fluorescent carbon dots with richer physical and chemical properties, greatly expanding their potential in the biomedical field. Nonetheless, fabricating carbon dots with integrated functionality for diagnostic and therapeutic modalities remains challenging. Herein, we develop a simple strategy to prepare Gd/Ru bimetallic doped fluorescent carbon dots (Gd/Ru-CDs) via a one-step microwave-assisted method with Ru(dcbpy)3Cl2, citric acid, polyethyleneimine, and GdCl3 as precursors. Multiple techniques were employed to characterize the morphology and properties of the obtained carbon dots. The Gd/Ru-CDs are high mono-dispersity, uniform spherical nanoparticles with an average diameter of 4.2 nm. Moreover, X-ray photoelectron spectroscopy (XPS), and Fourier transform infrared (FTIR) confirmed the composition and surface properties of the carbon dots. In particular, the successful doping of Gd/Ru enables the carbon dots not only show considerable magnetic resonance imaging (MRI) performance but also obtain better fluorescence (FL) properties, especially in the red emission area. More impressively, it has low cytotoxicity, excellent biocompatibility, and efficient reactive oxygen species (ROS) generation ability, making it an effective imaging-guided tumor treatment reagent. In vivo experiments have revealed that Gd/Ru-CDs can achieve light-induced tumor suppression and non-invasive fluorescence/magnetic resonance bimodal imaging reagents to monitor the treatment process of mouse tumor models. Thus, this simple and efficient carbon dot manufacturing strategy by doping functional metals has expanded avenues for the development and application of multifunctional all-in-one theranostics.

Development of a novel in vitro model for non-cavitated carious lesion formation reflecting carious lesion activity.

BACKGROUND: Oral biofilms are a critical component in dental caries formation. However, current remineralization studies often overlook the impact of microbial factors. Therefore, a comprehensive clinically relevant assessment of caries is needed. This study aimed to develop a novel in vitro model capable of generating non-cavitated carious lesions that incorporates both mineral loss and microbial activity using quantitative light-induced fluorescence-digital (QLF-D) technology. METHODS: A total of 44 artificial early carious lesions were formed using bovine incisors. The extent of fluorescence loss (ΔF) was analyzed using a QLF-D camera. Oral microcosm biofilms were then employed to construct 22 active and 22 inactive carious lesions. The red fluorescence emission rate (ΔR) and bacterial viability (RatioG/G+R) was measured using QLF-D camera and a live-dead bacterial assay, respectively. Independent t-tests were performed to compare ΔF, ΔR, and bacterial viability of artificial carious lesions according to their activity status. RESULTS: No significant difference in ΔF between the lesions was found based on activity status (p = 0.361). However, the ΔR of active lesions was 1.82 times higher than that of inactive lesions, and the RatioG/G+R was 1.49 times higher in active lesions than in inactive lesions (both p < 0.001). CONCLUSIONS: The significant differences observed in ΔR and RatioG/G+R between active and inactive lesions emphasize the importance of considering lesion activity status when evaluating the potential efficacy of remineralization agents. This study presents a novel in vitro remineralization assessment model that reflects carious lesion activity while controlling baseline mineral distributions of lesions.

Dual-state emission, mechanofluorochromism, and lipid droplet imaging of asymmetric D-π-A-D'-type triads.

Dual-state emission (DSE) is an emerging phenomenon wherein organic luminescent molecules display bright emissions in both molecularly isolated and packed states, addressing the challenge associated with the traditional paradigm of dyes with mono-state emission. This study presents the design and synthesis of two unsymmetrical triads, TPCA and TPCT, featuring a D-π-A-D' electronic structure by integrating phenothiazines, triphenylamines, and cyanostilbene. Photophysical assessments reveal that both molecules serve as robust DSEgens, exhibiting strong emissions in both solution and solid phases. TPCA displays ΦTHF 53.2% and Φsolids 43.2%, while TPCT exhibits ΦTHF 49.6% and Φsolids 37.5%. However, due to differences in molecular conformation and packing, they diverge in solid-state emission wavelengths and mechanofluorochromic behavior. In the solid state, TPCA emits strong red fluorescence, contrasting with TPCT, which emits orange fluorescence. Furthermore, TPCA demonstrates significant mechanofluorochromism (MFC), shifting from yellow to yellow-red upon mechanical grinding, while TPCT exhibits negligible MFC owing to conformational distinctions. As robust and low-toxic bioimaging agents, both TPCA and TPCT prove highly effective for lipid-droplet imaging studies. This research contributes valuable insights to the evolving field of DSE materials, elucidating the promising applications and mechanisms governing their versatile emission behaviors.

Red fluorescence intensity as a criterion for assessing remineralization efficacy in early carious lesions.

OBJECTIVES: Evaluating early carious lesion activity with an objective and clinically valid approach is crucial for developing effective treatment plans. Therefore, we here assessed the activity of non-cavitated carious lesions using a quantitative light-induced fluorescence-digital (QLF-D) camera and compared the remineralization efficiency after fluoride treatment according to the lesion's activity level. METHODS: Red fluorescence emission rate (ΔR) and fluorescence loss (ΔF) were evaluated in 44 non-cavitated carious lesions by using a QLF-D camera. Based on the ΔR level, the lesions were classified into 22 active (ΔR ≥37.55) and 22 inactive carious lesions (ΔR <37.55). Each lesion was treated with 1.23 % fluoride gel for 60 s and then immersed into artificial saliva for 7 days. Subsequently, ΔR and ΔF changes in the lesions were measured. RESULTS: Significant interactions between lesion activity and time were found for both ΔR and ΔF (p < 0.001). ΔR of active lesions declined faster and ΔF increased more steeply than did inactive lesions. Specifically, on day 7 post-fluoride treatment, the ΔR reduction rate was 1.40-times higher in active lesions, and the ΔF recovery rate was 2.50-times higher, indicating that active lesions respond more markedly to fluoride application. CONCLUSIONS: This study highlighted the significance of ΔR in predicting remineralization efficiency in non-cavitated carious lesions after fluoride application. It underscored the importance of accurately assessing caries activity when formulating effective treatment plans. Lesion activity, as determined by ΔR, not only influences the outcome of remineralization treatments but also provides a more objective measure for tailoring caries management strategies.

Red-Fluorescent Covalent Organic Framework Nanospheres for Trackable Anticancer Drug Delivery.

Covalent organic frameworks (COFs) have emerged as promising drug carriers due to their structural variability, inherent porosity, and customizable functions. However, most COFs used in drug delivery suffer from low cellular bioavailability and poor luminescence properties. In this study, we designed a series of size-tunable, crystalline, and red-fluorescent COF nanospheres (COFNSs) for trackable anticancer drug delivery. The semiconducting COFNSs were prepared by condensations of 1,3,5-triformylbenzene (TFB) with various dihydrazide blocks through the Schiff-base reaction, resulting in red emission at 647 nm and excellent fluorescence stability (∼100% for 1 h). Such fluorescence property allowed for systematic investigation of the cellular endocytosis pathway of COFNSs, visualization of drug delivery, and observation of the cell apoptosis process. The COFNSs exhibited high cell viability (>90%), a loading capacity of 183 wt % for the anticancer drug camptothecin (CPT), and significant enhancement in inhibiting 4T1 cancers both in vitro and in vivo as the CPT nanocarrier. This progress presents a valuable approach to design COF nanocarriers with integrated fluorescent and drug delivery functions.

Congo Red Staining in Digital Pathology: The Streamlined Pipeline for Amyloid Detection Through Congo Red Fluorescence Digital Analysis.

Renal amyloidosis is a rare condition caused by the progressive accumulation of misfolded proteins within glomeruli, vessels, and interstitium, causing functional decline and requiring prompt treatment due to its significant morbidity and mortality. Congo red (CR) stain on renal biopsy samples is the gold standard for diagnosis, but the need for polarized light is limiting the digitization of this nephropathology field. This study explores the feasibility and reliability of CR fluorescence on virtual slides (CRFvs) in evaluating the diagnostic accuracy and proposing an automated digital pipeline for its assessment. Whole-slide images from 154 renal biopsies with CR were scanned through a Texas red fluorescence filter (NanoZoomer S60, Hamamatsu) at the digital Nephropathology Center of the Istituto di Ricovero e Cura a Carattere Scientifico San Gerardo, Monza, Italy, and evaluated double-blinded for the detection and quantification through the amyloid score and a custom ImageJ pipeline was built to automatically detect amyloid-containing regions. Interobserver agreement for CRFvs was optimal (k = 0.90; 95% CI, 0.81-0.98), with even better concordance when consensus-based CRFvs evaluation was compared to the standard CR birefringence (BR) (k = 0.98; 95% CI, 0.93-1). Excellent performance was achieved in the assessment of amyloid score overall by CRFvs (weighted k = 0.70; 95% CI, 0.08-1), especially within the interstitium (weighted k = 0.60; 95% CI, 0.35-0.84), overcoming the misinterpretation of interstitial and capsular collagen BR. The application of an automated digital pathology pipeline (Streamlined Pipeline for Amyloid detection through CR fluorescence Digital Analysis, SPADA) further increased the performance of pathologists, leading to a complete concordance with the standard BR. This study represents an initial step in the validation of CRFvs, demonstrating its general reliability in a digital nephropathology center. The computational method used in this study has the potential to facilitate the integration of spatial omics and artificial intelligence tools for the diagnosis of amyloidosis, streamlining its detection process.

Real-time optical detection of endodontic infection using bacterial autofluorescence.

OBJECTIVES: For successful root canal treatment (RCT), it is essential to objectively assess the presence and activity of bacteria in the root canal system. However, current methods rely on subjective observations of root canal exudates. This study aimed to confirm whether real-time optical detection using bacterial autofluorescence can evaluate endodontic infection status by assessing the red fluorescence (RF) detected from root canal exudates. METHODS: During RCT, endodontic paper points were used to collect root canal exudates scored using conventional organoleptic tests to assess the severity of root canal infections. RF on the paper points was assessed using quantitative light-induced fluorescence (QLF) technology. RF intensity and area from the paper points were quantified, and their correlations with infection severity were assessed using their organoleptic scores. The oral microbiome composition of RF samples was compared with non-red fluorescent (non-RF) samples. RESULTS: The RF detection rate was nil and >98% in the non-infectious and severe groups. The RF intensity and area significantly increased with infection severity (p<0.001) and showed strong correlations with organoleptic scores (r=0.72, 0.82, respectively). The diagnostic accuracy for detecting root canal infection using RF intensity was good to excellent (AUC = 0.81-0.95) and increased with infection severity. The microbial diversity of the RF samples was significantly lower than that of the non-RF samples. Gram-negative anaerobic bacteria such as Prevotella and Porphyromonas were more predominant in RF samples. CONCLUSIONS: Optical detection using bacterial autofluorescence can objectively evaluate endodontic infection status in real-time by assessing the RF of endodontic root canal exudates. CLINICAL SIGNIFICANCE: This real-time optical technology can be utilised to detect endodontic bacterial infection without conventional incubation, allowing clinicians to determine the endpoint of chemomechanical debridement and increase the positive outcomes of RCTs.

Quantifying metal-binding specificity of CcNikZ-II from Clostridium carboxidivorans in the presence of competing metal ions.

Many proteins bind transition metal ions as cofactors to carry out their biological functions. Despite binding affinities for divalent transition metal ions being predominantly dictated by the Irving-Williams series for wild-type proteins, in vivo metal ion binding specificity is ensured by intracellular mechanisms that regulate free metal ion concentrations. However, a growing area of biotechnology research considers the use of metal-binding proteins in vitro to purify specific metal ions from wastewater, where specificity is dictated by the protein's metal binding affinities. A goal of metalloprotein engineering is to modulate these affinities to improve a protein's specificity towards a particular metal; however, the quantitative relationship between the affinities and the equilibrium metal-bound protein fractions depends on the underlying binding mechanisms. Here we demonstrate a high-throughput intrinsic tryptophan fluorescence quenching method to validate binding models in multi-metal solutions for CcNikZ-II, a nickel-binding protein from Clostridium carboxidivorans. Using our validated models, we quantify the relationship between binding affinity and specificity in different classes of metal-binding models for CcNikZ-II. We further illustrate the potential relevance of data-informed models to predicting engineering targets for improved specificity.

An ultra-small fluorescence zero-valent iron nanoclusters selectively kill gram-positive bacteria by promoting reactive oxygen species generation.

A list of the most dangerous bacteria that are multiple-drug resistance has been published by WHO, among which are various Gram-positive bacteria related with serious healthcare and community-associated infection. An effort is called for developing new strategies to combat the resistance, and nanomaterial-based approaches provide an ideal potential to mitigate the antimicrobial resistance as an alternative to antibiotics. Nanoscale zero-valent iron particles exhibited a good antimicrobial activity by triggering Fenton reaction, however, no zero-valent iron nanoclusters are developed as antimicrobial medical materials. In this work, a novel ultra-small zero-valent iron nanoclusters (usZVIN) was synthesized by one-step reduction in aqueous solutions, which exhibited bright red fluorescence at 616 nm. Interestingly, the usZVIN displayed an excellent selectively antibacterial activity against Gram-positive bacteria, and little effects on Gram-negative bacteria. The killing efficiency of usZVIN against S. aureus can reach 100 % with a concentration of 40 µg mL-1 after 1 h incubation, whereas there is no killing effect of usZVIN against E.coli even with a concentration of 900 µg mL-1 for 4 h. The antimicrobial mechanism of usZVIN was demonstrated to be the intracellular reactive oxygen species (ROS) production triggered by usZVIN due to its excellent peroxidase-like activity. Collectively, our findings suggested that usZVIN is a good medical-material candidate for fighting against Gram-positive bacterial infections, especially when we need leave beneficial Gram-negative bacteria intact.

A red fluorescence probe for reversible detection of HSO3-/H2O2 and its application in food samples and bioimaging.

Reducing agent SO2 and oxidant H2O2 are two essential substances in cells, and the balance between them is closely related to the survival of cells. SO2 derivative HSO3- is often used as food additive. Therefore, simultaneous detection of SO2 and H2O2 is of great significance in biology and food safety. In this work, we successfully developed a mitochondria-targeted red fluorescent probe (HBTI), which has excellent selectivity, high sensitivity and large Stokes shift (202 nm). HBTI and HSO3-/SO32- undergo Michael addition on the unsaturated C=C bond, and the addition product (HBTI-HSO3-) can react with H2O2 to restore the conjugated structure. Fluorescence changes from red to non-emissive and then restores to red, and can be detected quickly and visually. In addition, HBTI has been successfully targeted mitochondria, and achieved dynamic reversible response to SO2/H2O2 in living cells, and has been successfully applied to detect SO2 in food samples.

Multifunctional carbon dots for glutathione detection and Golgi imaging.

Glutathione (GSH) is present in almost every cell in the body and plays various integral roles in many biological processes. The Golgi apparatus is a eukaryotic organelle for the biosynthesis, intracellular distribution, and secretion of various macromolecules; however, the mechanism of GSH in the Golgi apparatus has not been fully elucidated. Here, specific and sensitive sulfur-nitrogen co-doped carbon dots (SNCDs) with orange-red fluorescence was synthesized for the detection of GSH in the Golgi apparatus. The SNCDs have a Stokes shift of 147 nm and excellent fluorescence stability, and they exhibited excellent selectivity and high sensitivity to GSH. The linear response of the SNCDs to GSH was in the range of 10-460 µM (LOD = 0.25 µΜ). More importantly, we used SNCDs with excellent optical properties and low cytotoxicity as probes, and successfully realized golgi imaging in HeLa cells and GSH detection at the same time.

Lymphatic Alterations Under Tattoos: Preliminary Reports of One Observational Study.

Background: The number of people within the European population having at least one tattoo has increased notably and with it the number of tattoo-associated clinical complications. The injected inks are known to be removed by the lymphatic vessels and can be found in the draining lymph nodes. Aim of the Study: To report our observations on the lymphatic drainages seen under tattoos using near infrared fluorescence imaging of these lymphatic vessels after the injection of indocyanine green. Material and Methods: Indocyanine green was injected intradermally at the basis of the 20 tattooed area(s) in 19 subjects (nine women and ten men; mean age = 28.6). Ten subjects had only black tattoos (six upper limbs and four lower limbs), five (two upper limbs and three lower limbs) black and white tattoos and five multi-colored tattooed limbs (four lower limbs and one upper limb). Results: The imaging exams revealed alterations in eight individuals, seven of whom had tattoos on their lower limbs. Furthermore, the imaging results showed that the abnormalities might be related to the tattooed limb, the tattoo extent and colour. Conclusion: Alterations of the cutaneous lymphatic channels are frequently observed under tattooed territories. Their causal factors should be more precisely studied in future works and these lymphatic alterations should be considered in tattooed patients when using similar imaging techniques for therapeutic and surgical assessments.

Single-molecule characterization of a bright and photostable deep-red fluorescent squaraine-figure-eight (SF8) dye.

Squaraine Figure Eight (SF8) dyes are a unique class of deep-red fluorescent dyes with self-threaded molecular architecture that provides structural rigidity while simultaneously encapsulating and protecting the emissive fluorochrome. Previous cell microscopy and bulk phase studies of SF8 dyes indicated order of magnitude enhancements in photostability over conventional pentamethine cyanine dyes such as Cy5. Studies conducted at the single molecule level now reveal that these ensemble level enhancements carry over to the single molecule level in terms of enhanced emission quantum yields, longer times to photobleaching, and enhanced total photon yields. When compared to Cy5, the SF8-based dye SF8(D4)2 possesses a three-fold larger single molecule emission quantum yield, exhibits order of magnitude longer average times before photobleaching, and exhibits twenty times larger photon yields. Additional features such as water solubility, fluorochrome encapsulation to protect it against nucleophilic attack, and selective biomarker targeting capability make SF8-based dyes promising candidates for biological labeling and microscopy applications and single molecule tracking.

Quantitative Assessment of Full Size Microplastics in Bottled and Tap Water Samples in Hong Kong.

Human exposure to microplastics (MPs) through drinking water has drawn serious concern recently because of the potential adverse health effects. Although there are reports on the occurrence of MPs in bottled water, little is known about the abundance of a whole spectrum of MPs with sizes ranging from 1 µm to 5 mm due to the restrictions of conventional MPs detection methods. Some studies using micro-Raman spectroscopy can achieve MPs with a size of <10 µm, however, quantitation of all MPs was extremely time consuming and only a small portion (<10%) of MPs would be analyzed. The present study quantified MPs from nine brands of bottled water using fluorescence microscopy and flow cytometry for MPs with a size of ≥50 µm and a size of <50 µm, respectively. The average abundance of MPs with a size of ≥50 µm in bottled water samples was found ranging from 8-50 particles L-1, while MPs with a size of <50 µm were found to be 1570-17,817 particles L-1, where the MPs abundance from mineral water samples were significantly more than distilled and spring water samples. The modal size and shape of MPs were found at 1 µm and fragments, respectively. Besides, three tap water samples obtained locally were analyzed and compared with the bottled water samples, where less MPs were found in tap water samples. In addition, contamination of MPs from bottle and cap and interference by addition of mineral salts were studied, where no significant difference from all these processes to the control sample was found, suggesting the major contamination of MPs was from other manufacturing processes. Estimated daily intake (EDI) of MPs increased substantially when data of small MPs are included, suggesting that previously reports on exposure of MPs from drinking water might be underestimated, as only large MPs were considered.