Fig. 4: Practical use of descSPIM in the drug discovery and 3D pathology.

a An example of Trastuzumab-administered whole cell-line derived xenograft (CDX; BT-474 human breast cancer cell line) imaging with fine axial (FA) mode, tilling light-sheet (TLS) method, flat-field correction (FFC) adoption, and multi-directional stack registration and fusing. Anti-CD31 antibody was also perfused before sampling. PI was stained during clearing. DL650 (Trastuzumab), PI, and FITC (CD31) were excited with 647 nm, 515 nm, and 488 nm laser beams, respectively. Using the θ stage, image tiles from the angles 0° and 180° were obtained. The two-sided stacks were then aligned and merged with ANTs and BigStitcher. Orthogonal cross-sections near the center of the specimen are shown as max intensity projection (MIP) images of a thickness of approximately 70 µm. Although validation of the imaging results by other methods is required, the descSPIM’s potential to acquire apparent drug distributions is anticipated to be beneficial for drug discovery and development. Voxel size: 3.45 × 3.45 × 10 µm³. The original grayscale 8-bit maps were pseudo-colored with a Blue, Yellow or Magenta look-up table (LUT). The imaging experiment was performed at twice with nearly identical results. b An example of 2 mm-thick CDX imaging with FA mode, TLS method, and FFC adoption. The sample was stained with PI and NHS-Alexa FluorⓇ 647 before clearing, and false-colored hematoxylin and eosin colors (3D Fluo-HE) were applied. Alexa FluorⓇ 647 and PI were excited with 647 nm and 515 nm laser beams, respectively. The data demonstrates that descSPIM has the potential to be used routinely in 3D clinical pathology examinations. Voxel size: 3.45 × 3.45 × 10 µm³. The original grayscale 8-bit maps were pseudo-colored with a Blue, Green or Fluo-HE LUT. The imaging experiment was performed twice with nearly identical results.