AIE-based fluorescent probe designed with xanthone as a π-bridge for detecting of ClO- in pericarp and living cells.

The role of ClO- in the physiological functioning of organisms is significant. In this paper, the four fluorescent probes HONx (HON1, HON2, HON3 and HON4) were prepared based on oxyanthracene through the introduction of different substituents, and their photophysical properties were investigated, among which the AIE effect of HON1 was the most significant, and therefore the fluorescent "turn-off" ClO- probe HON1-CN was chosen to be prepared by constructing the ClO- recognition site hydrazone bond at HON1. The ClO- recognises the hydrazone group in the probe HON1-CN, and when the hydrazone bond is broken, the aldehyde group is released, generating HON1 with yellow fluorescence. The probe HON1-CN is highly selective and stable for the detection of ClO- with a detection limit of 0.48 µM and a more than 10-fold increase in fluorescence intensity when the fluorescence is 'switched on', and to a lesser extent, the probe is also very good for the detection of hypochlorite ClO- in the pericarp. Finally, HON1-CN has also been used to detect the presence of ClO- in HeLa cells and zebrafish.

Sensing features, on-site detection and bio-imaging application of a tripodal tris(hydroxycoumarin) based probe towards Cu2+/His.

A new tripodal tris(hydroxycoumarin) based Schiff base, HCTN was synthesized and characterized by FT-IR, 1H NMR, 13C NMR and ESI-HRMS. The probe, HCTN exhibits cyan emission in DMSO/HEPES buffer (9:1, v/v) which selectively detects Cu2+ ion via turn-off fluorescence. The quenching of the fluorescence was due to the binding of the probe, HCTN towards paramagnetic Cu2+ ion resulting in chelation enhanced quenching effect (CHEQ). From the spectroscopic results, the limit of detection of Cu2+ ion was obtained as very low as 0.40 × 10-9 M. The complexation of the metal ion, Cu2+ towards the probe HCTN was confirmed by the ESI-HRMS and Job's plot analysis which supports 1:1 binding stochiometric ratio. In order to validate the affinity of Cu2+ ion towards histidine, the HCTN+Cu2+ system was utilized for the detection of histidine via turn-on mode by the metal displacement approach. The detection limit of His was found to be 7.31 × 10-10 M. In addition to the above, the probe was utilized for various detection applications such as paper strips, cotton swabs, logic gates and thin film applications. The probe, HCTN extends its application to the confocal bioimaging to sense the Cu2+ and Histidine intracellularly.

Synthesis and optical properties of tyramine-functionalized boron dipyrromethene dyes for cell bioimaging.

Tyramine signaling amplification (TSA) technology is generally applied in immunofluorescence, enzyme-linked immunoassays, in situ hybridization techniques, etc. Successful amplification of fluoresence signals cannot be achieved without excellent fluorescent dyes. BODIPY fluorophore is an ideal probe for cell fluorescence imaging, but pristine BODIPY cannot be direct used in the TSA system. In the paper, the new red-shifted tyramide-conjugated BODIPY (BDP-B/C/D) was synthesized via the Knoevenagel condensation reaction, which based on the tyramide-conjugated BODIPY (BDP-A). The synthesized dyes were combined with tyramine to obtain which could be used as a fluorescent substrate for enzymatic reaction of TSA. By using the selected substrate (BDP-C) in TSA, we found it to be more sensitive than the commercial dye 594 styramide for the detection of low-abundance antigen proteins.

A dual-channel fluorescent probe targeting lysosomes for differential detection of Cys/Hcy and GSH: Applications in food, pharmaceutical analysis and bioimaging.

Thiols function as antioxidants in food, prolonging shelf life and enhancing flavor. Moreover, thiols are vital biomolecules involved in enzyme activity, cellular signal transduction, and protein folding among critical biological processes. In this paper, the fluorescent probe PYL-NBD was designed and synthesized, which utilized the fluorescent molecule pyrazoline, the lysosome-targeted morpholine moiety, and the sensing moiety NBD. Probe PYL-NBD was tailored for the recognition of biothiols through single-wavelength excitation, yielding distinct fluorescence emission signals: blue for Cys, Hcy, and GSH; green for Cys, Hcy. Probe PYL-NBD exhibited rapid reaction kinetics (<10 min), distinct fluorescence response signals, and low detection limits (15.7 nM for Cys, 14.4 nM for Hcy, and 12.6 nM for GSH). Probe PYL-NBD enabled quantitative determination of Cys content in food samples and L-cysteine capsules. Furthermore, probe PYL-NBD had been successfully applied for confocal imaging with dual-channel detection of biothiols in various biological specimens, including HeLa cells, zebrafish, tumor sections, and Arabidopsis thaliana.

Autonomous multicolor bioluminescence imaging in bacteria, mammalian, and plant hosts.

Bioluminescence imaging has become a valuable tool in biological research, offering several advantages over fluorescence-based techniques, including the absence of phototoxicity and photobleaching, along with a higher signal-to-noise ratio. Common bioluminescence imaging methods often require the addition of an external chemical substrate (luciferin), which can result in a decrease in luminescence intensity over time and limit prolonged observations. Since the bacterial bioluminescence system is genetically encoded for luciferase-luciferin production, it enables autonomous bioluminescence (auto-bioluminescence) imaging. However, its application to multiple reporters is restricted due to a limited range of color variants. Here, we report five-color auto-bioluminescence system named Nano-lanternX (NLX), which can be expressed in bacterial, mammalian, and plant hosts, thereby enabling auto-bioluminescence in various living organisms. Utilizing spectral unmixing, we achieved the successful observation of multicolor auto-bioluminescence, enabling detailed single-cell imaging across both bacterial and mammalian cells. We have also expanded the applications of the NLX system, such as multiplexed auto-bioluminescence imaging for gene expression, protein localization, and dynamics of biomolecules within living mammalian cells.

A simple benzothiazolium-based sensor for cyanide detection: Applications in environmental analysis and bioimaging.

A new sensor based on Ethylbenzothiazolium-2-hydroxynaphthaldehyde conjugate-based fluorescent sensor, (E)-3-ethyl-2-(2-(2-hydroxynaphthalen-1-yl) vinyl) benzo[d]thiazol-3-ium iodide (SU-1) was designed and synthesized. The structure of SU-1 was confirmed by 1H NMR, 13C NMR, HRMS, and single crystal XRD spectral analysis. SU-1 displayed a colorimetric and fluorometric response in a DMSO:H2O (1:1,v/v) matrix, changing color from pale yellow to colorless visible to the naked eye, accompanied by a âˆ¼ 120 nm red-shift in the absorption spectra upon CN- addition. This shift, due to formation of deprotonation followed by the nucleophilic attack on the benzothiazolium ring's double bond, disrupts π-conjugation, blocking intramolecular charge transfer within SU-1. However, competitive anions showed negligible interference while detecting CN-. The Limit of detection for CN- was determined to be 0.27 nM, significantly below the WHO's permissible CN- concentration in drinking water (1.9 µM). Job's plot analysis shows that the binding stoichiometry of SU-1 to CN- is a 1:1, with a stability constant (Ka) of 1.58 x 104 M-1. The sensor demonstrated practical applications in environmental water samples and fluorescence imaging of intracellular CN- in CAD cell line.

PRAME expression in fibrosarcomatous dermatofibrosarcoma protuberans.

PRAME (PReferentially expressed Antigen in MElanoma) was first identified as a malignant melanoma-specific antigen. Recently, a few cases of fibrosarcomatous dermatofibrosarcoma protuberans (FS-DFSP) were shown to have positivity for PRAME, while conventional dermatofibrosarcoma protuberans (C-DFSP) was negative. Because PRAME may be of diagnostic utility in FS-DFSP and is raising expectations as a new immunotherapy target, we examined the positivity of PRAME in FS-DFSP. Twenty-one cases of FS-DFSP and age/sex/location-matched cases of C-DFSP as a control group were examined by immunohistochemistry for CD34 and PRAME. The results were then evaluated by H-score, which was objectively and semi-quantitatively calculated using the open-source bioimaging analysis software QuPath. The results revealed that the PRAME H-score in FS-DFSP was significantly higher than that in C-DFSP (p = 0.0137). As for CD34, the H-score in FS-DFSP was significantly lower than that in C-DFSP (p < 0.001). Using these two immunohistochemical analyses in combination, the sensitivity and specificity for the diagnosis of FS-DFSP were 86% and 90%, respectively. Double staining of CD34 and PRAME revealed that PRAME-positive and CD34-positive areas did not overlap. This is the largest study to examine PRAME expression in FS-DFSP, and it confirmed the usefulness of PRAME in diagnosing this condition.

Automated early ovarian cancer detection system based on bioinformatics.

Ovarian cancer is a common gynecological tumor, with a high mortality rate and difficult clinical treatment. Early detection of ovarian cancer has significant diagnostic value. In response to the problem of poor diagnostic performance of traditional early diagnosis methods, this article designed an automated early ovarian cancer detection system to improve the detection of early ovarian cancer. The conventional early diagnosis methods include serum CA125 (carbohydrate antigen 125) detection and positron emission tomography/computed tomography (PET/CT) imaging. This article combined serum CA125 detection and PET/CT imaging to detect the CA125 level and maximum standardized uptake value (SUV) in patient's serum. When the CA125 level exceeded 35U/ml and the maximum SUV value exceeded 2.5, the test was considered positive. This article selected 200 patients from Jingzhou Hospital for the experiment and compared the three detection methods. The average specificity of single serum CA125 detection, single PET/CT imaging, and automated detection in patients under 50 were 61.24%, 79.57%, and 97.79%, respectively. The automated early ovarian cancer detection system designed in this article can significantly improve the specificity of early ovarian cancer detection and has excellent application value for early ovarian cancer detection.

Optical Transparency Windows in Near-Infrared and Short-Wave Infrared for the Skin, Skull, and Brain: Fluorescence Bioimaging Using PbS Quantum Dots.

Fluorescence imaging (FI) employing near-infrared (NIR) light within the range of ~750-1350 nm enables biomedical imaging several millimeters beneath the tissue surface. More recent investigations into the short-wave IR (SWIR) transparency windows between ~1550-1870 and 2100-2300 nm highlight their superior capabilities. This research presents a comparison of IR-FI of PbS quantum dots, emitting at 990, 1310, and 1580 nm, through the mouse scalp skin, skull, and brain. The SWIR fluorescence is the most effectively transmitted signal, showing particularly significant enhancement when passing through the skull, which causes high light scattering. For the analysis of the imaging results and light propagation through the organs, their spectra of attenuation, absorption, and scattering coefficients are measured. In view of biomedical imaging, attenuation due to light scattering is a more destructive factor. Hence, the spatial resolution and imaging contrast can be improved by operating in SWIR due to decreased light scattering.

A high content imaging assay for identification of specific inhibitors of native Plasmodium liver stage protein synthesis.

Plasmodium parasite resistance to antimalarial drugs is a serious threat to public health in malaria-endemic areas. Compounds that target core cellular processes like translation are highly desirable, as they should be capable of killing parasites in their liver and blood stage forms, regardless of molecular target or mechanism. Assays that can identify these compounds are thus needed. Recently, specific quantification of native Plasmodium berghei liver stage protein synthesis, as well as that of the hepatoma cells supporting parasite growth, was achieved via automated confocal feedback microscopy of the o-propargyl puromycin (OPP)-labeled nascent proteome, but this imaging modality is limited in throughput. Here, we developed and validated a miniaturized high content imaging (HCI) version of the OPP assay that increases throughput, before deploying this approach to screen the Pathogen Box. We identified only two hits; both of which are parasite-specific quinoline-4-carboxamides, and analogs of the clinical candidate and known inhibitor of blood and liver stage protein synthesis, DDD107498/cabamiquine. We further show that these compounds have strikingly distinct relationships between their antiplasmodial and translation inhibition efficacies. These results demonstrate the utility and reliability of the P. berghei liver stage OPP HCI assay for the specific, single-well quantification of Plasmodium and human protein synthesis in the native cellular context, allowing the identification of selective Plasmodium translation inhibitors with the highest potential for multistage activity.