Tomocube

Research on organoids, the 3D cell aggregates replicating organ structures and functions, benefits significantly from the detailed and noninvasive imaging capabilities of Holotomography (HT). HT efficiently captures the dynamic behavior of cells within living organoids, including mitotic events, cyst formation, and crypt budding. Real-time, high-resolution observations with available quantitative measurements facilitated by HT are crucial for exploring the complex dynamics and morphology of the organoids.

In combination with the HT-ready 96-well plate, which supports a broad spectrum of cell types from 2D to 3D cultures, the HT-X1 is an ideal tool for leveraging the potential of organoids in drug development. It enables high-throughput screenings for rapid assessment of drug response and its toxicity on patient-derived tissues.

A recent study highlighted a successful example of such efforts, demonstrating how HT has set a new standard for comprehensive and rigorous statistical assessments for the biological studies of organoids (Lee et al., 2023). With HT, intricate subcellular structures of individual cells in small intestinal organoids were observed in intricate detail. Long-term monitoring of various biological processes within the specimen, including cell mitosis, migration, apoptosis, and other subcellular dynamics, was achieved effortlessly without the need for labeling or other time-consuming preparations.

Additionally, the observation of morphological changes was combined with quantitative parameters, including organoid volume, protein concentration, and mass, to effectively and comprehensively evaluate the response of the organoids to chemotherapy drug treatments. The study suggested that the Tomocube HT-X1 imaging system is an indispensable tool for organoid-based pharmacological screening applications.

While 2D cell culture systems have been instrumental in advancing our understanding of cell biology, they fall short in replicating the natural microenvironment of tissues due to the lack of cellular communication and interaction. Therefore, it is essential to study cell and tissue behavior in a 3D context that mirrors the structure of the extracellular matrix (ECM).

Among various 3D cell culture techniques, organ-on-a-chip is in increasing demand across diverse biomedical research fields. Unlike less advanced 3D culture systems, such as protein-based ECM or hydrogels, organ-on-a-chip combines cell culture with microfluidic devices (µFDs). By simplifying and miniaturizing integrated biological processes on microscale chips, it better imitates the dynamic mechanical properties and biochemical functionalities of living organs.

The HT-X1 is a powerful tool that offers numerous benefits for organ-on-a-chip research. It provides a precise representation of cellular behavior in a 3D environment with high resolution and in real-time, enabling researchers to better understand cell interactions in complex environments. Additionally, the HT-X1 can track cellular behaviors without relying on staining techniques, which is particularly useful when fluorescent staining is challenging in intricate 3D platforms.

The excellent ability of Tomocube low-coherence HT was demonstrated in an experiment observing live Hep3B cells on a 3D collagen type I and fiber structure within the DAX-1 chip (video). High-resolution images of live Hep3B cells were obtained, featuring detailed structures of organelles such as the nucleus, nucleolus, lipid droplets, mitochondria, and collagen fibers at exceptional spatial resolution. The Hep3B cells exhibited a distinct round-shape morphology that differs from their morphology in a 2D environment, highlighting the significant influence of the ECM on cell behavior and morphology.

Study tissues provides a more comprehensive and accurate understanding of biological processes, disease mechanisms, and therapeutic responses compared to studying isolated cells. However, research involving tissues presents numerous challenges. These include technical difficulties such as maintaining tissue integrity, manipulation, and culture, as well as technological limitations in imaging and analyzing tissue heterogeneity and structural complexity.

Holotomography (HT) is a pivotal advancement in live tissue imaging. It optimizes tissue research by offering deep structural insight of intact tissue sample without the need for arduous preparation steps such as thin tissue sectioning, fixation, and staining, which often compromise the morphological and physiological information obtainable from the specimen.

A recent review highlights the inevitable transition from 2D to 3D spatial profiling in cancer research, showcasing how HT has transformed our understanding of cancer cells and their surrounding tumor microenvironments (Wang et al., 2024). Tomocube’s 3D imaging technology addresses challenge such as extensive sample preparation, preservation of tissue's natural state, and post-imaging multimodal molecular data collection. These innovations are paving the way for the future of personalized medicine, enabling more precise and effective cancer diagnosis and treatment.

The video on the right showcases HT imaging capabilities, revealing remarkable spatial features of three different mouse reproductive tissues. Observations included the dynamics of live placental tissue, stages of spermatogenesis within seminiferous tubules, and detailed structures in oviduct tissue. Using HT, the morphology of epithelial cells, sperm cells, cilia cells, secretory cells, and the lamina propria were all visualized in high detail and in 3D. The timelapse capture of internal flow and beating in placental tissue offers additional insights into reproductive processes.