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In this study, we brought together X-ray induced photodynamic therapy (X-PDT) and chemo-drug (5-FU) for the treatment on colorectal cancer cells. This was achieved by developing a lipid-polymer hybrid nanoparticle delivery system (FA-LPNPs-VP-5-FU). It was prepared by incorporating a photosensitizer (verteporfin), chemotherapy drug (5-FU) and a targeting moiety (folic acid) into one platform. The average size of these nanoparticles was around 100 nm with low polydispersity. When exposed to clinical doses of 4 Gy X-ray radiation, FA-LPNPs-VP-5-FU generated sufficient amounts of reactive oxygen species, triggering the apoptosis and necrosis pathway of cancer cells. Our combined X-PDT and chemo-drug strategy was effective in inhibiting cancer cells' growth and proliferation. Cell cycle analyses revealed that our treatment induced G2/M and S phase arrest in HCT116 cells. Our results indicate that this combined treatment provides better antitumour effect in colorectal cancer cells than each of these modalities alone. This may offer a novel approach for effective colorectal cancer treatment with reduced off-target effect and drug toxicity.

Pain is currently assessed using subjective measurements, often not aligning with clinical symptoms. Therefore, objective pain level assessments, using minimally-invasive and molecular methods, are needed to assess disease activity and response to treatment in osteoarthritis and rheumatoid arthritis. We report sophisticated quantitative biochemical “signatures” from the label-free hyperspectral imaging (HSI) of cartilage tissue for the characterization of molecular composition, structure and functional status. Further study on sinuvium tissue provides evidence that HSI could be used as a novel technique to delineate disease state. Additionally, HSI could be used to objectively separate individuals based on pain severity providing molecular correlates of pain

Type 1 diabetes occurs when insulin secreting beta cells in pancreatic islets are destroyed leading to elevated glucose and ill health. Islet transplantation is an effective therapy, but islets are often damaged by the isolation process resulting in numerous, repeated transplants to achieve insulin independence. We have applied hyperspectral microscopy to damaged islets and have shown through the assessment of native cell autofluorescence we can detect heterogenous forms of damage (elevated ROS, inflammatory signalling and warm ischemia) in mouse islets. This approach has great potential to be translated clinically to minimise the burden of suboptimal islet transplantations.

Detecting reactive oxygen species (ROS) that play a critical role as redox modulators and signalling molecules in biological systems currently requires invasive methods such as ROS-specific indicators for imaging and quantification. We developed a non-invasive, real-time, label-free imaging technique for assessing the level of ROS in live cells and thawed cryopreserved tissues that is compatible with invivo imaging.The (AFMI) system used in this study is based on an adapted IX83 Olympus microscope as previously reported. It provides narrow excitation wavelength ranges (±5nm) from high power LEDs and several filter cubes to produce defined spectral regions, which span the excitation (340 nm–510 nm) and emission (420 nm–650 nm) wavelength ranges. In total, 18 distinct spectral channels (Nch= 18) are available and the samples are imaged in all these channels

Stem cell exhaustion plays a major role in the ageing of different tissues. Similarly, in vitro cell ageing during expansion prior to their use in regenerative medicine can severely compromise stem cell quality through progressive declines in differentiation and growth capacity. We utilized non-destructive multispectral assessment of native cell autofluorescence to investigate the metabolic mechanisms of in vitro mesenchymal stem cell (MSC) ageing in human bone marrow MSCs over serial passages (P2–P10). The spectral signals for NAD(P)H, flavins and protein-bound NAD(P)H were successfully isolated using Robust Dependent Component Analysis (RoDECA). NAD(P)H decreased over the course of hMSC ageing in absolute terms as well as relative to flavins (optical redox ratio). Relative changes in other fluorophore levels (flavins, protein-bound NAD(P)H) suggested that this reduction was due to nicotinamide adenine dinucleotide depletion rather than a metabolic shift from glycolysis to oxidative phosphorylation. Using multispectral features, which are determined without cell fixation or fluorescent labelling, we developed and externally validated a reliable, linear model which could accurately categorize the age of culture-expanded hMSCs. The largest shift in spectral characteristics occurs early in hMSC ageing. These findings demonstrate the feasibility of applying multispectral technology for the non-invasive monitoring of MSC health in vitro.

© 2019 Early detection of seeds, as a precursor tool for the prevention of Grass Seed Infestation, is analysed. It is demonstrated that terahertz time-domain spectroscopy and imaging techniques can be utilized to identify seeds in sheep-wool.

Background: Cell cycle analysis is important for cancer research. However, available methodologies have drawbacks including limited categorisation and reliance on fixation, staining or transformation. Multispectral analysis of endogenous cell autofluorescence has been shown to be sensitive to changes in cell status and could be applied to the discrimination of cell cycle without these steps. Methods: Cells from the MIA-PaCa-2, PANC-1, and HeLa cell lines were plated on gridded dishes and imaged using a multispectral fluorescence microscope. They were then stained for proliferating cell nuclear antigen (PCNA) and DNA intensity as a reference standard for their cell cycle position (G1, S, G2, M). The multispectral data was split into training and testing datasets and models were generated to discriminate between G1, S, and G2 + M phase cells. A standard decision tree classification approach was taken, and a two-step system was generated for each line. Results: Across cancer cell lines accuracy ranged from 68.3% (MIA-PaCa-2) to 73.3% (HeLa) for distinguishing G1 from S and G2 + M, and 69.0% (MIA-PaCa-2) to 78.0% (PANC1) for distinguishing S from G2 + M. Unmixing the multispectral data showed that the autofluorophores NADH, FAD, and PPIX had significant differences between phases. Similarly, the redox ratio and the ratio of protein bound to free NADH were significantly affected. Conclusions: These results demonstrate that multispectral microscopy could be used for the non-destructive, label free discrimination of cell cycle phase in cancer cells. They provide novel information on the mechanisms of cell-cycle progression and control, and have practical implications for oncology research.

To investigate the effect of maternal MitoQ treatment on renal disorders caused by maternal cigarette smoke exposure (SE). We have demonstrated that maternal SE during pregnancy increases the risk of developing chronic kidney disease (CKD) in adult offspring. Mitochondrial oxidative damage contributes to the adverse effects of maternal smoking on renal disorders. MitoQ is a mitochondria-targeted antioxidant that has been shown to protect against oxidative damage-related pathologies in many diseases. Female Balb/c mice (8 weeks) were divided into Sham (exposed to air), SE (exposed to cigarette smoke) and SEMQ (exposed to cigarette smoke with MitoQ supplemented from mating) groups. Kidneys from the mothers were collected when the pups weaned and those from the offspring were collected at 13 weeks. Maternal MitoQ supplementation during gestation and lactation significantly reversed the adverse impact of maternal SE on offspring's body weight, kidney mass and renal pathology. MitoQ administration also significantly reversed the impact of SE on the renal cellular mitochondrial density and renal total reactive oxygen species in both the mothers and their offspring in adulthood. Our results suggested that MitoQ supplementation can mitigate the adverse impact of maternal SE on offspring's renal pathology, renal oxidative stress and mitochondrial density in mice offspring.

© The Authors, published by EDP Sciences, 2017. Epitaxial Al x Ga 1-x Sb layers on GaSb and GaAs substrates have been grown by atmospheric pressure metalorganic chemical vapor deposition using TMAl, TMGa and TMSb. Nomarski microscope and a profiler were employed to examine the surface morphology and growth rate of the samples. We report the effect of growth temperature and V/III flux ratio on growth rate and surface morphology. Growth temperatures in the range of 520°C and 680°C and V/III ratios from 1 to 5 have been investigated. A growth rate activation energy of 0.73 eV was found. At low growth temperatures between 520 and 540°C, the surface morphology is poor due to antimonide precipitates associated with incomplete decomposition of the TMSb. For layers grown on GaAs at 580°C and 600°C with a V/III ratio of 3 a high quality surface morphology is typical, with a mirror-like surface and good composition control. It was found that a suitable growth temperature and V/III flux ratio was beneficial for producing good AlGaSb layers. Undoped AlGaSb grown at 580°C with a V/III flux ratio of 3 at the rate of 3.5 μm/hour shows p-Type conductivity with smooth surface morphology.

Intravital multiphoton microscopy has emerged as a powerful technique to visualize cellular processes in-vivo. Real time processes revealed through live imaging provided many opportunities to capture cellular activities in living animals. The typical parameters that determine the performance of multiphoton microscopy are speed, field of view, 3D imaging and imaging depth; many of these are important to achieving data from in-vivo. Here, we provide a full exposition of the flexible polygon mirror based high speed laser scanning multiphoton imaging system, PCI-6110 card (National Instruments) and high speed analog frame grabber card (Matrox Solios eA/XA), which allows for rapid adjustments between frame rates i.e. 5 Hz to 50 Hz with 512 x 512 pixels. Furthermore, a motion correction algorithm is also used to mitigate motion artifacts. A customized control software called Pscan 1.0 is developed for the system. This is then followed by calibration of the imaging performance of the system and a series of quantitative in-vitro and in-vivo imaging in neuronal tissues and mice.

Here we propose a region-recognition approach with iterative thresholding, which is adaptively tailored to extract the appropriate region or shape of spatial frequency. In order to justify the method, we tested it with different samples and imaging conditions (different objectives). We demonstrate that our method provides a useful method for rapid imaging of cellular dynamics in microfluidic and cell cultures.

The biological behaviour in terms of cellular incubation and organelle specificity for two complexes of the type fac-[Re(CO)3(phen)L], where phen is 1,10-phenanthroline and L is either 3-pyridyltetrazolate or4-cyanophenyltetrazolate, are herein investigated. The emission signal detected from the live insect Drosophila and human cell lines, generated by exploiting two-photon excitation at 830 nm to reduce cellular damage and autofluorescence, suggests photophysical properties that are analogous to those measured from dilute solutions, meaning that the complexes remain intact within the cellular environment. Moreover, the rhenium complex linked to 4-cyanophenyltetrazolateshows high specificity for the lipid droplets, whereas the complex bound to 3-pyridyltetrazolate tends to localise within the lysosomes. This differential localisation implies that in thesecomplexes, organelle specificity can be achieved and manipulated by simple functional group transformations thus avoiding more complex bioconjugation strategies. More importantly, these results highlight the first example of phosphorescent labeling of the lipid droplets, whose important cellular functions have been recently highlighted along with the fact that their role in the metabolism of healthy and diseased cells has not been fully elucidated.

Stacked self-assembled In₀.₅Ga₀.₅As∕GaAs quantum dot infrared photodetectors grown by low-pressure metal-organic chemical vapor deposition, with and without silicon dopants in the quantum dot layers, are investigated. The increase of dark currents observed at higher doping levels is attributed to higher defect density leading to stronger sequential resonant tunneling and to lowering of the operating temperature of the device.

A comprehensive study of the morphological, optical and microstructural properties of mass-transport (MT) and conventionally grown GaN by hydride vapour-phase epitaxy is presented. Spatially resolved techniques have been utilized to reveal in a comparative way, the characteristics of the material grown either in predominant vertical or lateral growth modes. A strong donor-acceptor pair (DAP) emission is observed from the MT regions with a distinctive intensity contrast between the exciton and DAP emission bands from MT and nontransport regions. Secondary ion mass spectroscopy and imaging were employed to investigate the impurity incorporation into different regions. An increase of residual oxygen and aluminium impurity concentrations was found in the MT areas. In addition, positron annihilation spectroscopy showed a strong signal of Ga vacancy clusters in the MT grown material. The increase of the point defect concentrations of both Ga vacancy and oxygen impurity, most likely forming defect complexes, is related to the enhancement of the DAP emission., QC 20120907