Understanding light intensity and temperature increase is of considerable importance in designing or performing in vivo optogenetic experiments. Our study describes the optimal light power at target depth in the rodent brain that would maximize activation of light-gated ion channels while minimizing temperature increase. Monte Carlo (MC) simulations of light delivery were used to provide a guideline for suitable light power at a target depth. In addition, MC simulations with the Pennes bio-heat model using data obtained from measurements with a temperature-measuring cannula having 12.3 mV/°C of thermoelectric sensitivity enabled us to predict tissue heating of 0.116 °C/mW on average at target depth of 563 μm and specifically, a maximum mean plateau temperature increase of 0.25 °C/mW at 100 μm depth for 473 nm light. Our study will help to improve the design and performance of optogenetic experiments while avoiding potential over- and under-illumination.

Alzheimer's disease (AD) is driven by complex interactions between vascular dysfunction, glymphatic system impairment, and neuroinflammation. Vascular aging, characterized by arterial stiffness and reduced cerebral blood flow (CBF), disrupts the pulsatile forces necessary for glymphatic clearance, exacerbating amyloid-beta (Aβ) accumulation and cognitive decline. This review synthesizes insights into the mechanistic crosstalk between these systems and explores their contributions to AD pathogenesis. Emerging machine learning (ML) tools, such as DeepLabCut and Motion sequencing (MoSeq), offer innovative solutions for analyzing multimodal data and enhancing diagnostic precision. Integrating ML with imaging and behavioral analyses bridges gaps in understanding vascular-glymphatic dysfunction. Future research must prioritize these interactions to develop early diagnostics and targeted interventions, advancing our understanding of neurovascular health in AD.

In microscopic imaging of biological tissues, particularly real-time visualization of neuronal activities, rapid acquisition of volumetric images poses a prominent challenge. Typically, two-dimensional (2D) microscopy can be devised into an imaging system with 3D capability using any varifocal lens. Despite the conceptual simplicity, such an upgrade yet requires additional, complicated device components and usually suffers from a reduced acquisition rate, which is critical to properly document rapid neurophysiological dynamics. In this study, we implemented an electrically tunable lens (ETL) in the line-scan confocal microscopy (LSCM), enabling the volumetric acquisition at the rate of 20 frames per second with a maximum volume of interest of 315 × 315 × 80 µm³. The axial extent of point-spread-function (PSF) was 17.6 ± 1.6 µm and 90.4 ± 2.1 µm with the ETL operating in either stationary or resonant mode, respectively, revealing significant depth axial penetration by the resonant mode ETL microscopy. We further demonstrated the utilities of the ETL system by volume imaging of both cleared mouse brain ex vivo samples and in vivo brains. The current study showed a successful application of resonant ETL for constructing a high-performance 3D axially scanning LSCM (asLSCM) system. Such advances in rapid volumetric imaging would significantly enhance our understanding of various dynamic biological processes.

We describe a new method for recording spatially incoherent common-path off-axis color digital holograms. We present the theoretical and experimental evidence to demonstrate an incoherent common-path off-axis color digital holographic (ICOCH) system capable of capturing information from three-dimensional color objects under incoherent illumination, both in transmission and reflection modes. Fresnel incoherent correlation holography (FINCH), a common-path system, is a frequently used incoherent holography technique. Our proposed system is conceptually similar to an advanced form of FINCH; moreover, it has three advantages over this advanced form of FINCH. First, removal of the spatial light modulator makes our system simpler and more cost-effective. Second, removal of the polarizer or analyzer allows for greater light throughput. Third, the off-axis optical configuration enables separation of zero-order and twin images with only a single exposure per color rather than requiring three exposures per color for in-line holography FINCH. Therefore, we believe that this simple and cost-effective system with high light throughput can acquire incoherent holograms for different colors involving single exposure for each color, which makes the ICOCH system suitable for many applications.

We propose a Monte Carlo (MC) method based on a direct photon flux recording strategy using inhomogeneous, meshed rodent brain atlas. This MC method was inspired by and dedicated to fibre-optics-based optogenetic neural stimulations, thus providing an accurate and direct solution for light intensity distributions in brain regions with different optical properties. Our model was used to estimate the 3D light intensity attenuation for close proximity between an implanted optical fibre source and neural target area for typical optogenetics applications. Interestingly, there are discrepancies with studies using a diffusion-based light intensity prediction model, perhaps due to use of improper light scattering models developed for far-field problems. Our solution was validated by comparison with the gold-standard MC model, and it enabled accurate calculations of internal intensity distributions in an inhomogeneous near light source domain. Thus our strategy can be applied to studying how illuminated light spreads through an inhomogeneous brain area, or for determining the amount of light required for optogenetic manipulation of a specific neural target area.