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Genetically encoded optical sensors and advancements in microscopy instrumentation and techniques have revolutionized the scientific toolbox available for probing complex biological processes such as release of specific neurotransmitters. Most genetically encoded optical sensors currently used are based on fluorescence and have been highly successful tools for single-cell imaging in superficial brain regions. However, there remains a need to develop new tools for reporting neuronal activity in vivo within deeper structures without the need for hardware such as lenses or fibers to be implanted within the brain. Our approach to this problem is to replace the fluorescent elements of the existing biosensors with bioluminescent elements. This eliminates the need of external light sources to illuminate the sensor, thus allowing deeper brain regions to be imaged noninvasively. Here, we report the development of the first genetically encoded neurotransmitter indicators based on bioluminescent light emission. These probes were optimized by high-throughput screening of linker libraries. The selected probes exhibit robust changes in light output in response to the extracellular presence of the excitatory neurotransmitter glutamate. We expect this new approach to neurotransmitter indicator design to enable the engineering of specific bioluminescent probes for multiple additional neurotransmitters in the future, ultimately allowing neuroscientists to monitor activity associated with a specific neurotransmitter as it relates to behavior in a variety of neuronal and psychiatric disorders, among many other applications.

BACKGROUND: Positron emission tomography (PET) has had a transformative impact on oncological and neurological applications. However, still much of PETs potential remains untapped with limitations primarily driven by low spatial resolution, which severely hampers accurate quantitative PET imaging via the partial volume effect (PVE). PURPOSE: We present experimental results of a practical and cost-effective ultra-high resolution brain-dedicated PET scanner, using our depth-encoding Prism-PET detectors arranged along a compact and conformal gantry, showing substantial reduction in PVE and accurate radiotracer uptake quantification in small regions. METHODS: The decagon-shaped prototype scanner has a long diameter of 38.5 cm, a short diameter of 29.1 cm, and an axial field-of-view (FOV) of 25.5 mm with a single ring of 40 Prism-PET detector modules. Each module comprises a 16 × 16 array of 1.5 × 1.5 × 20-mm3 lutetium yttrium oxyorthosillicate (LYSO) scintillator crystals coupled 4-to-1 to an 8 × 8 array of silicon photomultiplier (SiPM) pixels on one end and to a prismatoid light guide array on the opposite end. The scanners performance was evaluated by measuring depth-of-interaction (DOI) resolution, energy resolution, timing resolution, spatial resolution, sensitivity, and image quality of ultra-micro Derenzo and three-dimensional (3D) Hoffman brain phantoms. RESULTS: The full width at half maximum (FWHM) DOI, energy, and timing resolutions of the scanner are 2.85 mm, 12.6%, and 271 ps, respectively. Not considering artifacts due to mechanical misalignment of detector blocks, the intrinsic spatial resolution is 0.89-mm FWHM. Point source images reconstructed with 3D filtered back-projection (FBP) show an average spatial resolution of 1.53-mm FWHM across the entire FOV. The peak absolute sensitivity is 1.2% for an energy window of 400-650 keV. The ultra-micro Derenzo phantom study demonstrates the highest reported spatial resolution performance for a human brain PET scanner with perfect reconstruction of 1.00-mm diameter hot-rods. Reconstructed images of customized Hoffman brain phantoms prove that Prism-PET enables accurate radiotracer uptake quantification in small brain regions (2-3 mm). CONCLUSIONS: Prism-PET will substantially strengthen the utility of quantitative PET in neurology for early diagnosis of neurodegenerative diseases, and in neuro-oncology for improved management of both primary and metastatic brain tumors.

Nourished by academic research, venture capital investments, and a vibrant community, the bioeconomy is emerging as a transformational force in places such as Boston and San Francisco. Not surprisingly, today's [...]

Ammonia (NH3) has received significant attention recently as a promising carbon-free, low-cost, and safe candidate as a chemical energy storage element and/or fuel alternative. As for any new fuel, fundamental aspects of the combustion process, such as the detailed chemical kinetic pathways of NH3 oxidation, flame propagation and stability, and emissions, are to be understood to predict its combustion behavior in practical systems. One aspect of improving our understanding of the combustion behavior of NH3 is to explore the emission spectra of spherically expanding and/or Bunsen-type flames under controlled laboratory conditions. In this study, flame spectra from NH3-containing mixtures were obtained utilizing two different experimental flame configurations, namely a spherically propagating flame and a modified Mckenna burner flame. Two different fuel mixtures were investigated. The first was an oxy-ammonia mixture consisting of NH3 + (25% O2 +75% N2) with the goal of studying neat ammonia. The second mixture consisted of H2:NH3:N2 with a ratio of 45:40:15 by volume. Several excited-state species within the UV-visible spectral region were identified. For instance, NO* emission features were found between 220 and 280 nm. In addition, OH* emission was detected around 310 nm, and an NH* feature was seen around 337 nm. Moreover, a broadband intensity was reported over the visible range (400 ~ 800 nm), where several NH2* features were also detected. The effect of the mixture on the flame spectra was investigated by examining the two different mixtures utilizing the same experimental apparatus (spherical flame in this case), and it was shown that the mixture has an effect on the presence of NH2* features, which was also backed by chemical kinetics predictions for the NH2* mole fraction profiles. It was shown (from kinetics) that the oxy-ammonia mixture produced more NH2* than the H2:NH3:N2 mixture by up to 113%. Additionally, the effects of the flame configuration were also investigated by studying the same mixture (the H2:NH3:N2 mixture in this case), and it was revealed that the spectra produced using the modified Mckenna burner had less noise, and all features were easily identified. Lastly, laminar flame speeds of the oxy-ammonia mixture were measured using the spherical flame apparatus. The laminar flame speed followed the expected pattern, peaking near a 1.1 equivalence ratio with a measured flame speed of 22.7 cm/s. [ABSTRACT FROM AUTHOR]