Deep Learning-powered Bessel-beam Multi-parametric Photoacoustic Microscopy

Enabling simultaneous and high resolution quantification of the total concentration of hemoglobin (CHb), oxygen saturation of hemoglobin (sO2), and cerebral blood flow (CBF), multi parametric photoacoustic microscopy (PAM) has emerged as a promising tool for functional and metabolic imaging of the live mouse brain. However, due to the limited depth of focus imposed by the Gaussian beam excitation, the quantitative measurements become inaccurate when the imaging object is out of focus. To address this problem, we have developed a hardware-software combined approach by integrating Bessel beam excitation and conditional generative adversarial network (cGAN) based deep learning. Side by side comparison of the new cGAN powered Bessel-beam multi parametric PAM against the conventional Gaussian beam multi parametric PAM shows that the new system enables high resolution, quantitative imaging of CHb, sO2, and CBF over a depth range of ~600 μm in the live mouse brain, with errors 13 to 58 times lower than those of the conventional system. Better fulfilling the rigid requirement of light focusing for accurate hemodynamic measurements, the deep learning powered Bessel beam multi parametric PAM may find applications in large field functional recording across the uneven brain surface and beyond (e.g., tumor imaging).

Tools for increasing visual encounter probabilities for invasive species removal: a case study of brown treesnakes

Early detection and rapid response (EDRR) are essential to identifying and decisively responding to the introduction or spread of an invasive species, thus avoiding population establishment and improving the probability of achieving eradication. However, detection can be challenging at the onset of a species invasion as low population densities can reduce the likelihood of detection and conceal the true extent of the situation until the species is well established. This is doubly challenging if the invading species displays cryptic behavior or is nocturnal, thus further limiting opportunities for its discovery. Survey methods that maximize a searcher’s ability to detect an incipient population are therefore critical for successful EDRR. Brown treesnakes (Boiga irregularis) on Guåhan are a classic cautionary example of the dangers of not detecting an invasion early on, and the risk of their introduction to other islands within the Marianas, Hawai’i and beyond remains. Nocturnal visual surveys are known to detect brown treesnakes of all sizes and are the primary detection tool used by the Brown Treesnake Rapid Response Team, but detection probability remains low in complex forest habitats. As such, we investigated the use of two potential enhancements to nocturnal visual surveys – a live mouse lure and spray scent attractant – that may create hotspots of increased detection probability during surveys. We found that, while brown treesnake detection probabilities were low for all surveys, visual surveys conducted on transects with live mouse lures resulted in detection probabilities that were 1.3 times higher than on transects without live mouse lures. Conversely, the spray scent attractant did not increase the probability of detecting brown treesnakes compared to transects without scent, and in fact had detection probabilities that were 0.66 times lower, though the reasons for this phenomenon are unclear. Unlike scent attractants, live mouse lures likely provide both visual and olfactory cues that attract brown treesnakes to transects and thus provide more opportunities to detect and capture them. These enhancements were trialed on Guåhan, where prey populations are depressed. It remains unclear whether live mouse lures will be as effective for EDRR applications in prey-rich settings.

Tumor cell plasticity, heterogeneity, and resistance in crucial microenvironmental niches in glioma

Both the perivascular niche (PVN) and the integration into multicellular networks by tumor microtubes (TMs) have been associated with progression and resistance to therapies in glioblastoma, but their specific contribution remained unknown. By long-term tracking of tumor cell fate and dynamics in the live mouse brain, differential therapeutic responses in both niches are determined. Both the PVN, a preferential location of long-term quiescent glioma cells, and network integration facilitate resistance against cytotoxic effects of radiotherapy and chemotherapy—independently of each other, but with additive effects. Perivascular glioblastoma cells are particularly able to actively repair damage to tumor regions. Population of the PVN and resistance in it depend on proficient NOTCH1 expression. In turn, NOTCH1 downregulation induces resistant multicellular networks by TM extension. Our findings identify NOTCH1 as a central switch between the PVN and network niche in glioma, and demonstrate robust cross-compensation when only one niche is targeted.

A ratiometric electrochemical sensor for selectively monitoring monoamine oxidase A in the live brain

Herein, an electrochemical method for selectively sensing and accurately quantifying of monoamine oxidase A (MAO-A) in cortex and thalamus of live mouse brains was reported. Using this tool, it was...

Calcium Signaling in Pancreatic Immune Cells In situ

Immune cells were identified in intact live mouse pancreatic lobules and their Ca2+ signals, evoked by various agents, characterized and compared with the simultaneously recorded Ca2+ signals in neighboring acinar and stellate cells. Immunochemistry in the live lobules indicated that the pancreatic immune cells most likely are macrophages. In the normal pancreas the density of these cells is very low, but induction of acute pancreatitis (AP), by a combination of ethanol and fatty acids, markedly increased the number of the immune cells. The principal agent eliciting Ca2+ signals in the pancreatic immune cells was ATP, but these cells also frequently produced Ca2+ signals in response to acetylcholine and to high concentrations of bradykinin. Pharmacological studies, using specific purinergic agonists and antagonists, indicated that the ATP-elicited Ca2+ signals were mediated by both P2Y1 and P2Y13 receptors. The pancreatic immune cells were not electrically excitable and the Ca2+ signals generated by ATP were primarily due to release of Ca2+ from internal stores followed by store-operated Ca2+ entry through Ca2+ release-activated Ca2+ channels. The ATP-induced intracellular Ca2+ liberation was dependent on both IP3 generation and IP3 receptors. We propose that the ATP-elicited Ca2+ signal generation in the pancreatic immune cells is likely to play an important role in the severe inflammatory response to the primary injury of the acinar cells that occurs in AP.