LLSM Dataset for Virtual and Augmented reality

LLSM Dataset for Virtual and Augmented Reality. This dataset includes data generated by Lattice Light Sheet Microscopy. Image metadata is described in the file: FilesDescription_LLSM_Datasets_VR_AR.xlsx. This dataset includes: Rab5(early endosomes), Plasma Membrane CellMask (endosomal pathway) and CD63-Turquoise.2 (late-multivesicular endosomes). Lattice light-sheet microscopy. LLSM was done on a commercialized version of a previously described setup4 from 3i (Denver, USA). Cells were scanned incrementally through a 20 μm long light sheet in 500 nm steps using a fast piezoelectric flexure stage equivalent to ∼271 nm or 600 nm steps using a fast piezoelectric flexure stage equivalent to ∼325 nm, with respect to the detection objective and were imaged using a sCMOS camera (Orca-Flash 4.0; Hamamatsu, Bridgewater, NJ). Excitation was achieved with 488- (Coherent, Santa Clara, CA), 560- or 642-nm diode lasers (MPB Communications, Canada) through an excitation objective (Special Optics 28.6× 0.7 NA 3.74-mm water-dipping lens) and detected via a Nikon CFI Apo LWD 25× 1.1 NA water-dipping objective with a 2.5× tube lens with a final pixel size of 104 nm. Lattice light-sheet imaging was performed using an excitation pattern of outer NA equal to 0.55 and inner NA equal to 0.493. Acquired data were deskewed, a necessary step to realigned image frames, then deconvolved using LLSpy (https://github.com/tlambert03/LLSpy).面向虚拟现实与增强现实的LLSM（Lattice Light Sheet Microscopy，晶格光片显微镜）数据集。本数据集包含晶格光片显微镜采集得到的实验数据，图像元数据详见文件FilesDescription_LLSM_Datasets_VR_AR.xlsx。本数据集涵盖三类标记样本的数据：Rab5（早期内体）、细胞膜CellMask（内吞通路标记）以及CD63-Turquoise.2（晚期多泡内体）。本次实验采用美国丹佛3i公司商业化的、此前已有文献报道的setup4实验装置。细胞通过快速压电弯曲载物台，以500 nm步长（对应检测物镜方向约271 nm步长）或600 nm步长（对应检测物镜方向约325 nm步长），在20 μm长的光片内进行逐段扫描，并通过sCMOS（科学互补金属氧化物半导体）相机（Orca-Flash 4.0；滨松光子学株式会社，新泽西州布里奇沃特）完成成像。激发光源采用488 nm（相干公司，加利福尼亚州圣克拉拉）、560 nm或642 nm二极管激光器（加拿大MPB通信公司），通过激发物镜（Special Optics 28.6× 0.7 NA 3.74-mm水浸物镜）激发荧光；荧光信号经由尼康CFI Apo LWD 25× 1.1 NA水浸物镜搭配2.5×筒镜进行采集，最终成像像素尺寸为104 nm。本次晶格光片成像采用的激发模式为：外数值孔径（NA）0.55，内数值孔径（NA）0.493。采集得到的原始数据需先进行斜切校正（这是图像帧重对准的必要步骤），随后通过LLSpy工具（https://github.com/tlambert03/LLSpy）完成反卷积处理。