Fig. 2: Overview of 3D imaging by HiLoTRUST.
From: Rapid full-color serial sectioning tomography with speckle illumination and ultraviolet excitation
a Schematic of the HiLoTRUST system. b Workflow of the 3D imaging with HiLoTRUST, which contains six steps: (1) fluorescent staining with DAPI and PI for ~150 s, (2) widefield imaging with raster-scanning under speckle illumination by LD, (3) widefield imaging with raster-scanning under uniform illumination by two UV-LEDs, (4) shaving off the imaged layer with the vibratome to expose a layer underneath, (5) HiLo image processing, (6) 3D reconstruction by direct image stacking without image spatial registration. During imaging, the previous four steps will be repeated until the entire sample has been imaged and the whole procedure is fully automated with lab-built hardware and control programs. Note: TRUST and HiLoTRUST images have been normalized with min-max normalization for better visualization.
