Fluorescence micro-optical sectioning tomography using acousto-optical deflector-based confocal scheme

Abstract

Fluorescent labeling has opened up the possibility of clarifying the complex distribution and circuit wiring of specific neural circuits for particular functions. To acquire the brain-wide fluorescently labeled neural wiring, we have previously developed the fluorescence micro-optical sectioning tomography imaging system. This employs simultaneous mechanical sectioning and confocal imaging of the slices, and is capable of acquiring the image dataset of a centimeter-sized whole-mouse brain at a voxel resolution of [Formula: see text]. We analyze the key optical considerations for the use of an acousto-optical deflector (AOD) scanner-based confocal detection scheme in this system. As a result, the influence of confocal detection, the imaging site during sectioning, and AOD fast scan mode on signal-to-background noise ratio are described. It is shown that mechanical sectioning to separate the slice and optical sectioning by confocal detection should be combined to maximize background suppression in simultaneous fast scan imaging while sectioning system setup.

Denoising Across Data Acquisition Modalities for Mesoscopic Scale Optical Neuroimaging

“…The proposed GAN model is a mix-max game with the Wasserstein cost. The GAN model converting the denoised SI images to the noisy TDI images alike is shown in (3). The other GAN model is built alike in the denoised TDI images to the noisy SI images direction.…”

“…T HE mesoscopic scale optical imaging systems can capture images of the neuronal fibers, which may expand across a large part of the brain [1]. In this paper, we focus on the image denoising for the micro optical sectioning tomography (MOST) and its followup works [2], [3]. The imaging process and the result is illustrated in Fig.…”

Optical Brain Imaging: A Powerful Tool for Neuroscience

“…deflectors (Fig. 2) are able to rescue some of the information loss during sectioning and imaging [46], and help to achieve a higher resolution as well as a faster scan imaging [39,45]. Furthermore, to maintain the fluorescence signals, different embedding approaches during specimen preparation also have diverse effects on both the processes and results of the MOST system; these include glycol methacrylate, Unicryl, and LR White, among which modified glycol methacrylate is relatively more suitable [44].…”

“…Schematic of the fluorescence MOST system[21,39,40].A The system usually consists of a confocal microscope, an acoustical optical deflector scanner, and a microtome with a diamond knife and a moveable chamber. The motion of the chamber matches spatial coordinate axes.…”

Robust Cell Detection for Large-Scale 3D Microscopy Using GPU-Accelerated Iterative Voting

“…Knife-Edge Scanning Microscopy (KESM) is an optical imaging technique that allows researchers to quickly collect terabyte-scale 3D images by serially sectioning a sample (Mayerich et al, 2008 ) labeled using either traditional brightfield stains, such as thionine, Golgi-Cox, or hematoxylin and eosin (H&E), as well as transgenic fluorescent labels (Qi et al, 2015 ). KESM allows researchers to collect detailed images describing cell structure and vascular/neuronal connectivity across large (cm 3 ) volumes.…”