Rapid multicolor three-dimensional (3D) imaging for entire organs with high resolution remains a captivating scientific pursuit. A significant challenge associated with whole-organ imaging pertains to the laborious and time-consuming sample preparation and image acquisition.

To this end, we first proposed a fast, cost-effective, and robust multicolor whole-organ 3D imaging method assisted with ultraviolet (UV) surface excitation and vibratomy-assisted sectioning, termed translational rapid ultraviolet-excited sectioning tomography (TRUST). With an inexpensive UV light-emitting diode (UV-LED), TRUST achieves widefield fluorescence and autofluorescence imaging simultaneously while preserving low system complexity and system cost. By combining serial sectioning and real-time staining tech...[ Read more ]

Rapid multicolor three-dimensional (3D) imaging for entire organs with high resolution remains a captivating scientific pursuit. A significant challenge associated with whole-organ imaging pertains to the laborious and time-consuming sample preparation and image acquisition.

To this end, we first proposed a fast, cost-effective, and robust multicolor whole-organ 3D imaging method assisted with ultraviolet (UV) surface excitation and vibratomy-assisted sectioning, termed translational rapid ultraviolet-excited sectioning tomography (TRUST). With an inexpensive UV light-emitting diode (UV-LED), TRUST achieves widefield fluorescence and autofluorescence imaging simultaneously while preserving low system complexity and system cost. By combining serial sectioning and real-time staining techniques, TRUST has achieved automated 3D imaging with excellent staining uniformity and time efficiency. Its fast, robust, and high-content multicolor 3D imaging capability has been demonstrated by imaging all vital organs and two embryos. TRUST also demonstrates potential in the field of 3D histopathology, specifically for acquiring high-quality histological images of whole organs. Traditional methods in 3D histopathology are known for their time-consuming and laborious nature, often taking several weeks to complete. However, by integrating TRUST with deep learning, we have developed HistoTRUST, a pipeline that can rapidly and automatically generate virtual hematoxylin and eosin (H&E) stained histological images of whole organs with subcellular resolution. With HistoTRUST, we have successfully produced 3D histology images of all vital organs within 1–3 days. An inherent drawback of TRUST or HistoTRUST is its limited axial resolution, determined solely by the penetration depth of UV light. To this end, we have introduced a new 3D imaging system known as HiLoTRUST by integrating the TRUST system with the structure illumination method known as HiLo microscopy. HiLoTRUST presents improved imaging capabilities, particularly in terms of axial resolution. 3D imaging of diverse mouse organs and two human cancer specimens has been conducted with HiLoTRUST. The imaging results clearly demonstrate a remarkable enhancement in the optical sectioning capability compared to TRUST, validating the reliability and effectiveness of the HiLoTRUST approach.

In conclusion, this thesis introduces three promising platforms for rapid and cost-effective whole-organ high-resolution imaging while relieving researchers from the burden of sample preparation.