Tomocube

Organoids, the 3D cell aggregates replicating organ structures and functions, benefit significantly from HT's detailed and unlabeled imaging capabilities. It efficiently captures dynamic behavior of cells within living organoids, including mitotic events, epithelial cell dissection, cyst formation, and crypt budding. Real-time, high-resolution observations with available quantitative measurements facilitated by HT are crucial for scrutinizing 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, HT-X1 is an ideal tool to leverage the in vitro expandability of organoids to conduct high-throughput screenings for rapid assessment of drug response and its toxicity on tissues of individual patients.

In a recent study, HT was demonstrated to set a new standard for comprehensive, rigorous statistical assessments for the studies of organoids (Lee et al., 2023). With HT, intricate subcellular structures of individual cells in small intestinal organoids were observed in detail with a lateral resolution of 156 nm and axial resolution of 947 nm. Long-term monitoring of different biological processes within the specimen, such as cell apoptosis, migration, mitosis, and other subcellular dynamics, could be easily achieved as the procedures required no time-consuming preparation or labeling. In addition, 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 cisplatin treatment. The study suggested the Tomocube low-coherence HT imaging system as an indispensable tool for organoids in pharmacological screening applications.

As the traditional 2D cell culture systems cannot mimic the natural tissue microenvironment due to the lack of cellular communication and interaction, it is essential to investigate the behavior of cells and tissues within a 3D context that can imitate the structure of the extracellular matrix (EMC) of the tissue.

Among several 3D cell culture techniques, microfluidic devices (µFDs) are in increasing demand for diverse biomedical research fields because of their ability to simplify and miniaturize integrated biological processes on microscale chips. µFDs are also proven to better mimic the dynamic mechanical properties and biochemical functionalities of the whole living organs than less advanced 3D culture systems, such as protein-based EMC or hydrogels.

However, conventional techniques face challenges in imaging miniaturized samples and labeling live specimens within µFDs due to limitations stemming from channel complexity and light scattering problems.

HT-X1 is a powerful tool that offers various benefits for observing cell dynamics in a 3D channel by providing a precise representation of cellular behavior in a 3D environment in high resolution and in real-time, enabling researchers to understand better how cells interact and behave in complex environments. In addition, the HT-X1 can track cellular behaviors without the need for staining, which is useful when fluorescent staining is not easy for complicated 3D platforms. This capability allows researchers to observe cellular behavior without depending on staining techniques.

The excellent capturing ability of this low-coherence Holotomography system was demonstrated in an experiment aimed to observe live Hep3B cells residing on a 3D collagen type I and fiber structure within the DAX-1 chip.

High-resolution images of live Hep3B cells were achieved featuring detailed organelles structures including nucleus, nucleolus, lipid droplet, mitochondria, and collagen fiber at exceptional spatial resolution. Hep3B cells also displayed a distinct round-shape morphology that differs from their morphology in a 2D environment, highlighting the important influence of extracellular matrix (ECM) on cell behavior and morphology.