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SmartSPIM 組織層光掃描影像系統

SmartSPIM 層光掃描 3D 螢光影像系統 - Selective Plane Illumination Microscopy (SPIM), 是可導入 6 組選擇性波長的雷射激發光束, 光束是從側邊以水平面 ( selective plane ) 對澄清透明化的組織, 作類似電腦斷層掃描的光切照明, , 以取得透明組織的全貌 3D 重建螢光影像. 其好處在於大面積3D取樣, 快速與高解析.

組織樣本最大尺寸範圍 : 3 x 5.5 x 1.2 cm  (標準 2 x 2.5 x 1.2 cm )



傳統 共軛焦掃描顯微鏡 (Confocal Microscopy ) 是在鏡頭下的焦平面, 以 " 點方式 "  作為成像的基礎, 無法快速取得組織樣本的 3D 全貌影像.

SPIM 是是在鏡頭下的焦平面, 從組織樣本側邊, 雷射激雷射激發光束以水平面方向射出, 以一層一層的平面光束 ( selective plane ), 作為 " 面方式 " 成像的基礎, 可以快速3D成像.



 

 

 

Cleared with SmartClear II Pro and imaged with SmartSPIM.

Sample courtesy of G. Allan Johnson, Duke Center for In Vivo Microscopy.






Immunolabeled GFAP (cyan) and Beta-amyloid plaques (magenta) in a 5xFAD mouse brain model of AD

Images were acquired using the ASI 15X objective on SmartSPIM with 0.41 x 0.41 x 1 µm voxel size. A 100 µm MIP in the xy-plane (left) and a resliced 100 µm MIP in the xz-plane (right) from the same stack is shown, highlighting the nearly isotropic resolution. Sample courtesy of the Epp Lab at the University of Calgary.




Light sheet imaging system optimized for resolution, speed, and flexibility for large samples

SmartSPIM is an advanced light sheet microscope capable of imaging large tissue samples with high speed and resolution. The system was designed from the ground up, with no limitation based on commercially available microscope bodies. The unique illumination arm of the SmartSPIM creates a flat-top illumination profile laterally, and a focused beam axially. To achieve homogeneous axial point spread functions (PSFz) across the entire FOV, the illumination optics scan the beam axially while synchronized to the camera’s rolling shutter detection.

SmartSPIM offers the following:

Ability to use custom and commercially available objective

Custom illumination optics Zemax designed and optimized for achromatic performance across all points in the sweeping field of view

Detection optics Zemax designed and optimized for all current large format objectives and camera sensors

Focus compensation for RI mismatches and chromatic focal shifts

Compatible with both aqueous and organic clearing methods

Simple, user-friendly software interface

High speed acquisition: 20-40 minutes per channel per organ at single cell resolution, enabling the acquisition of dozens of datasets in a single week



 
 

Whole mouse brain processed with LifeCanvas’ tissue processing pipeline. Preserved with SHIELD, cleared with SmartClear II Pro, labeled with SmartLabel with anti-ChAT, and imaged with SmartSPIM at 4 µm Z-step and 1.8 µm XY pixel size. Tissue courtesy of the Fishell Lab at Broad Institute.



 
 

Mouse brain hemisphere displaying ChAT-positive neurons in both the brainstem and basal forebrain and dense labeling of Neuropeptide Y throughout the hypothalamus. Tissue was SHIELD perfused, cleared with SmartClear II Pro, and immunolabeled with SmartLabel with anti-NPY (cyan) and anti-ChAT (magenta). Imaged using SmartSPIM at 4 µm Z-step and 1.8 µm XY pixel size. Tissue courtesy of Hongwei Dong, Bin Zhang, and Neda Khanjani and the Center for Integrative Connectomics (CIC) lab.