KEYWORDS: Signal to noise ratio, Deep learning, Two photon imaging, Image enhancement, Imaging systems, Education and training, Two photon excitation microscopy, Signal processing, Neurons, Neural networks
We developed a high-speed two-photon volumetric imaging system with hundreds of axial layers that match a deep-learning denoising model to capture millisecond-level functional changes in individual neurons with high SNR. Compare with general deep-learning methods, the spatial information-based training method not only enhances SNR by 300% but prevents temporal distortion. Our proof-of-concept experiment focused on calcium dynamics in cerebellum Purkinje cells, revealing similar responses in the parallel dendritic layers, yet significant divergence in the somatic area. This sheds light on the intricate signal processing at individual neuron levels, validating our imaging system.
KEYWORDS: In vivo imaging, Laser scanners, Optical resolution, Photoacoustic microscopy, Brain, Whole body imaging, Surgery, Neuroimaging, Calcium, Animal model studies
Surgical removal of the head cuticle of Drosophila is necessary for optical brain imaging, leading to the Drosophila death in a short period of time and thus hindering long term monitoring. Targeting to the unmet need of surgery free procedure for Drosophila brain researches, in this study, laser scanning optical resolution photoacoustic microscopy (LSOR-PAM) of in vivo cuticle intact RCaMP labeled Drosophila mushroom body (MB) is presented. The MB is a higher-order olfactory center in Drosophila brain. This study paves the way toward exploring LSOR-PAM functional imaging capability of cuticle intact Drosophila brain under olfactory stimulus.
KEYWORDS: Brain, In vivo imaging, 3D image processing, 3D photoacoustic microscopy, Brain imaging, Laser scanners, Optical resolution, Photoacoustic microscopy, Confocal microscopy, Surgery
To study the structure and functions of the Drosophila brain, confocal microscopy is commonly used. However, surgical removal of the head cuticle of Drosophila is required because the cuticle hinders both the optical excitation and detection. Such invasive surgery may affect brain functions and prohibits long term monitoring. Targeting to the unmet need of surgery free procedure, here we propose laser scanning optical resolution photoacoustic microscopy (LSOR-PAM) for in vivo three dimensional cuticle intact Drosophila brain imaging. Cuticle intact Drosophila brains with cells in optic lobes expressing fluorescent protein DsRed, which serves as an optical absorber and thus a photoacoustic signal source, were imaged. Acquired in vivo 3D LSOR-PAM cuticle-intact brain images were cross-validated using their confocal microscopic counterparts with the cuticles being surgically removed. Acoustic and optical attenuation of the cuticles and degradation in spatial resolution caused by the cuticles were also measured, which explains the reason why LSOR-PAM outperforms confocal microscopy for cuticle intact brains. In addition, the optical absorption bleaching of DsRed expressing optic lobes as a function of the number of the repeated experiments was measured, verifying the LSOR-PAM long-term monitoring capability. In summary, we have demonstrated 3D LSOR-PAM of the Drosophila brain without invasive surgery for the first time. The focus of the future work will be on ways to explore its functional imaging capability on the cuticle intact Drosophila brain.
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