Fluorescent proteins with emission wavelengths in the near-infrared and infrared range are in high demand for whole-body imaging techniques. We report near-infrared fluorescent proteins eqFP650 and eqFP670. To our knowledge, eqFP650 is the brightest fluorescent protein with emission maximum above 635 nm, and eqFP670 displays the most red-shifted emission maximum and high photostability.

A simple method to evaluate the hemoglobin oxygen saturation and relative hemoglobin concentration in a tissue from diffuse reflectance spectra in the visible wavelength range is put forward in this paper. It was assumed that while oxygenated and deoxygenated hemoglobin contributions to light attenuation are strongly variable functions of wavelength, all other contributions to the attenuation including scattering are smooth wavelength functions and can be approximated by Taylor series expansion. Based on this assumption, a simple, robust algorithm suitable for real time monitoring of the hemoglobin oxygen saturation in the tissue has been derived. This algorithm can be used with different fiber probe configurations for delivering and collecting light passed through the tissue. An experimental technique using this algorithm has been developed for in vivo monitoring during artery occlusion and in vitro monitoring of blood samples. The experimental results obtained are presented in the paper.

We present a comprehensive analysis of the submissions to the first edition of the Endoscopy Artefact Detection challenge (EAD). Using crowd-sourcing, this initiative is a step towards understanding the limitations of existing state-of-the-art computer vision methods applied to endoscopy and promoting the development of new approaches suitable for clinical translation. Endoscopy is a routine imaging technique for the detection, diagnosis and treatment of diseases in hollow-organs; the esophagus, stomach, colon, uterus and the bladder. However the nature of these organs prevent imaged tissues to be free of imaging artefacts such as bubbles, pixel saturation, organ specularity and debris, all of which pose substantial challenges for any quantitative analysis. Consequently, the potential for improved clinical outcomes through quantitative assessment of abnormal mucosal surface observed in endoscopy videos is presently not realized accurately. The EAD challenge promotes awareness of and addresses this key bottleneck problem by investigating methods that can accurately classify, localize and segment artefacts in endoscopy frames as critical prerequisite tasks. Using a diverse curated multi-institutional, multi-modality, multi-organ dataset of video frames, the accuracy and performance of 23 algorithms were objectively ranked for artefact detection and segmentation. The ability of methods to generalize to unseen datasets was also evaluated. The best performing methods (top 15%) propose deep learning strategies to reconcile variabilities in artefact appearance with respect to size, modality, occurrence and organ type. However, no single method outperformed across all tasks. Detailed analyses reveal the

Conventional 3D multicellular tumor spheroids of head and neck squamous cell carcinoma (HNSCC) consisting exclusively of cancer cells have some limitations. They are compact cell aggregates that do not interact with their extracellular milieu, thus suffering from both insufficient extracellular matrix (ECM) deposition and absence of different types of stromal cells. In order to better mimic in vivo HNSCC tumor microenvironment, we have constructed a 3D stroma-rich in vitro model of HNSCC, using cancer-associated MeWo skin fibroblasts and FaDu pharynx squamous cell carcinoma. The expression of stromal components in heterospheroids was confirmed by immunochemical staining. The generated co-culture FaDu/MeWo spheroids were applied to study penetration, distribution and antitumor efficacy of photoactive drugs such as Temoporfin and Chlorin e6 used in the photodynamic therapy flow cytometry and fluorescence microscopy techniques. We also investigated the distribution of photodiagnostic agent Indocyanine Green. We demonstrated that the presence of stroma influences the behavior of photoactive drugs in different ways: (i) No effect on Indocyanine Green distribution; (ii) lower accumulation of Chlorin e6; (iii) better penetration and PDT efficiency of Temoporfin. Overall, the developed stroma-rich spheroids enlarge the arsenal of in vitro pre-clinical models for high-throughput screening of anti-cancer drugs.

Tetragonal xenotime-type yttrium orthophosphate (YPO4) Nd(3+) doped nanoparticles suitable for biomedical applications were prepared by microwave-hydrothermal treatment. We applied the energy transfer probing based on the analysis of kinetics of impurity quenching to determine the presence and spatial position of -OH fluorescence quenching acceptors in the impurity-containing nanoparticles. We show that the impurity quenching kinetics of the 0.1 at% Nd(3+) doped YPO4 nanoparticles is a two stage (ordered and disordered) static kinetics, determined by a direct energy transfer to the -OH acceptors. Analyzing the ordered stage, we assume that the origin of the -OH groups is the protonation of the phosphate groups, while analyzing the disordered stage, we assume the presence of water molecules in the mesopores. We determine the dimension of the space of the -OH acceptors as d = 3 and quantify their absolute concentration using the disordered Förster stage of kinetics. We use the late stage of kinetics of fluorescence hopping (CDD ≫ CDA) quenching (the fluctuation asymptotics) at 1 at% Nd(3+) concentration as an energy transfer probe to quantify the relative concentration of -OH molecular groups compared to an optically active rare-earth dopant in the volume of NPs, when energy migration over Nd(3+) donors to the -OH acceptors accelerates fluorescence quenching. In doing so we use just one parameter α = γ(A)/γ(D) = n(A)√[C(DA)]/n(D)√[C(DD)], defined by the relation of concentration of the -OH acceptors to the concentration of an optically active dopant. The higher is the α, the higher is the relative concentration of -OH acceptors in the volume of nanoparticles. We find α = 2.95 for the 1 at% Nd(3+):YPO4 NPs that, according to the equation for α, and the results obtained for the values of the microparameters CDD(Nd-Nd) = 24.6 nm(6) ms(-1) and CDA(Nd-OH) = 0.6 nm(6) ms(-1), suggests twenty times higher concentration for acceptors other than donors. As the main result we have established that the majority of -OH acceptors is located not on the surface of the Nd(3+):YPO4 nanoparticles, as many researchers assumed, but in their volume, and can be either associated with crystal structure defects or located in the mesopores.

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The COVID-19 pandemic has updated research on inactivation of coronaviruses with physical, chemical, and physical-chemical treatments. The review focuses on the inactivation of coronaviruses including the recent SARS-CoV-2 and viruses of other groups with analogous structure using optical radiation. The antiviral effects of different optical ranges from vacuum ultraviolet to infrared radiation are described in terms of the mechanisms of virus photoinactivation, sensitive molecular targets and efficacy. Direct and photosensitized damaging effects of light on the viral molecular structures are considered. Information on the applied pathogen photoinactivation technologies and the advantages of light sources for future applications is provided.